Zhang Di, Liu Jing, Zhang Xin, Bai Ling, Wang Heng, Guo Zhijie, Wei Liyu, Wang Sheng, Xu Daoming
DOI:10.13702/j.1000-0607.20251210
摘要:ObjectiveTo observe the effects of low-frequency pulsed electromagnetic fields (LF-PEMFs) combined with thunder fire moxibustion on muscle function and inflammatory factor levels in patients with primary osteoporosis (POP).MethodsPOP patients were randomly divided into LF-PEMFs group(30 cases,4 cases dropped out), thunder fire moxibustion group(30 cases,3 cases dropped out), and combination group(30 cases,3 cases dropped out). The LF-PEMFs group received treatment at a frequency of 16 Hz and an intensity of 20 mT (30 min/session, 3 times/week). The thunder fire moxibustion group received moxibustion at Mingmen (GV4), Yaoyangguan (GV3), and bilateral Ganshu (BL18), Shenshu (BL23), and Dachangshu (BL25) (30 min/session, 3 times/week). All patients received continuous treatment for 3 months. Assessments of 6-meter walking speed and serum inflammatory factors [interleukin (IL)-1β, IL-6, and tumor necrosis factor-α (TNF-α)] were conducted before treatment, after 3 months of treatment, and at a 6-month follow-up. Bone mineral density (BMD) was assessed before treatment and at the 6-month follow-up.ResultsAfter 3 months of treatment, compared with baseline, 6-meter walking speed significantly was increased in the LF-PEMFs, thunder fire moxibustion, and combination groups (P<0.05, P<0.01), and serum contents of IL-1β, IL-6, and TNF-α were significantly decreased in all three groups (P<0.01). At the 6-month follow-up, compared with baseline, 6-meter walking speed remained significantly higher in all groups (P<0.05, P<0.01), serum IL-6 and TNF-α contents were significantly decreased in all groups (P<0.05, P<0.01), and serum IL-1β was significantly decreased in the combination group (P<0.01). Compared with 3 months of treatment, the thunder fire moxibustion group showed further significant reductions in serum IL-1β and IL-6 contents at the 6-month follow-up (P<0.05). After 3 months of treatment, the combination group exhibited significantly higher 6-meter walking speed and lower serum TNF-α content compared with the LF-PEMFs group and the thunder fire moxibustion group (P<0.05, P<0.01),and lower serum IL-1β and IL-6 contents compared with the LF-PEMFs group (P<0.01, P<0.05). At the 6-month follow-up, the combination group showed higher 6-meter walking speed than the LF-PEMFs group and the thunder fire moxibustion group (P<0.01, P<0.05), lower serum IL-6 and TNF-α contents than both the LF-PEMFs group and the thunder fire moxibustion group (P<0.01, P<0.05), and lower serum IL-1β content than the thunder fire moxibustion group (P<0.01). There were no statistically significant differences in total lumbar and total hip BMD within or between groups before treatment and at the 6-month follow-up.ConclusionLF-PEMFs combined with thunder fire moxibustion can effectively improve muscle function, reduce serum inflammatory factor levels, and exert a positive effect on BMD in POP patients.
关键词:Primary osteoporosis;Thunder fire moxibustion;Low-frequency pulsed electromagnetic field;Muscle function;Inflammatory factor
Huang Xinyu, Su Shengyong, Huang Wenjing, Zheng Guangmei, Li Xin
DOI:10.13702/j.1000-0607.20251166
摘要:ObjectiveTo explore the effect of “qi-regulating” electroacupuncture on rats with cervical spondylotic radiculopathy (CSR) based on the protein kinase B (AKT)/glycogen synthase kinase-3β (GSK-3β)/nuclear factor erythroid 2-related factor 2 (Nrf2) signaling pathway, and to clarify its potential mechanism.MethodsMale SD rats were randomly divided into blank group, model group and electroacupuncture group, with 10 rats in each group. The CSR rat model was established by intraspinal catheter insertion into cervical vertebral canal. Starting on the 7th day after successful model establishment, rats in the electroacupuncture group received electroacupuncture at bilateral “Hegu”(LI4) and“Taichong”(LR3) for 20 min once a day for 7 consecutive days. Gait score, thermal withdrawal latency and mechanical pain threshold measured by Von Frey anesthesiometer were observed before and after intervention. HE staining was adopted to observe the pathological morphological changes of spinal cord tissue. ELISA was used to detect the contents of interleukin (IL)-1β, IL-6, tumor necrosis factor-α (TNF-α) and superoxide dismutase (SOD) in spinal cord tissue. Microassay method was used to detect the contents of malondialdehyde (MDA) and ferrous ion (Fe2+). DCFH-DA fluorescent probe was applied to detect the level of reactive oxygen species (ROS) in spinal cord tissue. Western blot was used to detect the ratios of phosphorylated (p)-AKT/AKT and p-GSK-3β/GSK-3β protein expression as well as the protein expression levels of Nrf2, glutathione peroxidase 4 (GPX4) and ferritin heavy chain 1 (FTH1) in spinal cord tissue.ResultsCompared with the blank group, the model group showed decreased mechanical pain threshold, shortened thermal withdrawal latency and elevated gait score (P<0.01). The contents of TNF-α, IL-1β, IL-6, MDA, Fe2+ and ROS level in spinal cord tissue were significantly increased (P<0.01), while the content of SOD, the ratios of p-AKT/AKT and p-GSK-3β/GSK-3β, as well as the protein expression levels of Nrf2, FTH1 and GPX4 were remarkably decreased (P<0.01). All above indexes were reversed in the electroacupuncture group compared with the model group (P<0.01, P<0.05).Conclusion“qi-regulating” electroacupuncture can alleviate pain in rats with cervical spondylotic radiculopathy, which may be realized by regulating the AKT/GSK-3β/Nrf2 signaling pathway in spinal cord tissue, reducing oxidative stress injury, suppressing ferroptosis and relieving inflammatory response in damaged nerve tissue.
Zhang Shuo, Cui Yang, Sun Zhong-Ren, Zhou Xinyu, Cao Yu, Wang Ze, Chen Yao, Yang Juandi, Dai Xueyuan, Yin Hongna
DOI:10.13702/j.1000-0607.20251170
摘要:Acupuncture has demonstrated significant therapeutic efficacy in the treatment and rehabilitation of ischemic stroke, effectively improving patients’ neurological function and daily living abilities. However, the underlying mechanisms by which it promotes neural repair have not yet been fully elucidated. This article systematically reviews and summarizes the research progress on acupuncture promoting neurogenesis after ischemic stroke. It elaborates on the biological basis of neurogenesis following ischemic stroke and its crucial role in functional recovery. The molecular mechanisms through which acupuncture promotes neurogenesis are deeply analyzed from the aspects of promoting neural stem cell proliferation and differentiation, regulating the expression of neurotrophic factors, and activating related signaling pathways. Additionally, the limitations of current research are analyzed and future directions are proposed, providing an important theoretical basis for further exploring the mechanism of acupuncture in promoting neurogenesis and developing new therapeutic strategies for stroke based on neural regeneration.
Wang Feiyan, Li Yue, Zeng Huijuan, Lin Shengshi, Dong Chunxuan, Ma Chongbing
DOI:10.13702/j.1000-0607.20260143
摘要:Structural and functional abnormalities in synaptic plasticity are closely related to the onset and progression of post-stroke depression (PSD). Acupuncture has been clinically recognized for its safety and efficacy in PSD intervention.This article analyzed the mechanisms of synaptic plasticity impairment underlying emotional disorders in PSD, and summarized the research progress on acupuncture intervention for PSD from the perspective of synaptic plasticity. Stroke-induced cerebral ischemia and hypoxia trigger energy metabolism disorders, excitotoxicity, and oxidative stress. Acupuncture can exert its antidepressant effects by modulating synaptic plasticity through multiple pathways, including inhibiting oxidative stress and ferroptosis, regulating neurotransmitter and receptor expression, activating brain‑derived neurotrophic factor and its related signaling pathways, modulating neuronal autophagy in neurons of key brain regions.
摘要:ObjectiveTo observe the effects of electroacupuncture (EA) intervention at different time points on cuproptosis-related proteins and Cu2+ levels in the ischemic penumbra of rats with cerebral ischemia-reperfusion injury (CIRI) after acute ischemic stroke (IS), so as to explore the mechanism by which acupuncture inhibits cuproptosis to improve neurological function in CIRI rats, as well as the therapeutic efficacy of intervention administered at different time windows.MethodsSD rats were randomly divided into sham operation group, model group, 0 h acupuncture group, 24 h acupuncture group and 48 h acupuncture group, with 12 rats in each group. The middle cerebral artery occlusion/reperfusion (MCAO/R) model was established. Rats in each EA group received 30 min of EA at “Baihui” (GV20) and “Fengfu” (GV16) at 0 h, 24 h and 48 h after reperfusion, respectively. Modified neurological severity score (mNSS) was used to evaluate the recovery of neurological deficit. 2,3,5-triphenyltetrazolium chloride (TTC) staining was adopted to detect cerebral infarct volume. Colorimetric assay was performed to measure Cu2+ content in brain tissues of the ischemic penumbra. Real-time quantitative PCR and Western blot were used to detect the mRNA and protein expression of ferredoxin 1 (FDX1), dihydrolipoamide S-acetyltransferase (DLAT), lipoic acid synthetase (LIAS), and solute carrier family 31 member 1 (SLC31A1) in the ischemic penumbra.ResultsCompared with the sham operation group, the model group exhibited significantly elevated mNSS score, cerebral infarct volume, Cu2+ content in ischemic penumbra tissues, as well as increased mRNA and protein expression of FDX1 and LIAS (P<0.01); meanwhile, the mRNA and protein expression of DLAT and SLC31A1 were markedly decreased (P<0.01). Compared with the model group, all acupuncture groups presented reduced mNSS score, cerebral infarct volume, Cu2+ level, together with down-regulated mRNA and protein expression of DLAT and SLC31A1 (P<0.01, P<0.05), and up-regulated mRNA and protein expression of FDX1 and LIAS (P<0.01, P<0.05). Compared with the 24 h acupuncture group and 48 h acupuncture group, the 0 h acupuncture group showed distinctly lower mNSS score, cerebral infarct volume, Cu2+ content, and weaker mRNA and protein expression of DLAT and SLC31A1 (P<0.01, P<0.05), accompanied by remarkably higher mRNA and protein expression of FDX1 and LIAS (P<0.01, P<0.05). The 24 h acupuncture group achieved better outcomes in cerebral infarct volume, Cu2+ content, and mRNA expression of DLAT, SLC31A1, FDX1 and LIAS relative to the 48 h acupuncture group (P<0.01, P<0.05).ConclusionEA intervention can effectively suppress cuproptosis and alleviate cerebral injury in CIRI rats, and early EA intervention exerts superior efficacy in ameliorating neurological impairment after CIRI. The underlying mechanism may involve regulating the expression of FDX1, DLAT, LIAS and SLC31A1, reducing brain Cu2+ concentration, and thereby inhibiting neuronal cuproptosis.
Zeng Yahan, Chen Xiaohong, Yu Zheng, Yu Shuguang, Yang Sha
DOI:10.13702/j.1000-0607.20260256
摘要:Primary dysmenorrhea (PD) is a common gynecological disorder affecting women of reproductive age worldwide, characterized by high prevalence, recurrent attacksand impaired quality of life. Acupuncture has definite therapeutic effects on PD, and related studies on its mechanism are increasingly deepening. The core pathogenesis of PD is the disordered brain-body bidirectional interaction. From the perspective of brain-body interaction, this paper systematically summarizes the mechanisms of acupuncture in intervening PD, including synchronously regulating central networks such as default mode network, sensorimotor network, and descending pain modulatory system, modulating signaling pathways including hypothalamic-pituitary-ovarian axis, hypothalamic-pituitary-adrenal axis, Toll-like receptor 4/nuclear factor-κB, and phosphatidylinositol 3‑kinase/protein kinase B/mammalian target of rapamycin, as well as improving uterine local pathological status, optimizing uterine blood flow, correcting prostaglandin imbalance, and relieving smooth muscle spasm. These effects synergistically block the brain-body vicious cycle of PD and achieve multi-system, multi-level holistic regulation and long-lasting analgesia.
Qu Zhengyang, Su Yangshuai, Liu Xiaoyu, Luo Min, Jing Xianghong
DOI:10.13702/j.1000-0607.20251317
摘要:ObjectiveTo observe the therapeutic effect of electroacupuncture (EA) at “Shangjuxu” (ST37) on colitis in mice, and to elucidate the mediating mechanism of the sacral parasympathetic cholinergic neuroimmune pathway.MethodsIn the first experiment, C57BL/6J mice were randomly assigned to a normal group, a model group, and an EA group,with 15 mice in each group. The colitis model was established by ad libitum administration of 2.5% dextran sodium sulfate solution for 7 days, while the normal group received plain drinking water. Mice in the EA group were subjected to EA stimulation at bilateral ST37 for 30 min per session under isoflurane anesthesia on days 1, 3, 5 and 7 of model induction. The body weight and colon length of mice in each group were measured, and the percentage change in body weight was calculated. HE staining was used to observe pathological alterations in colonic tissues. The MSD multiplex assay was adopted to detect colonic levels of C-X-C motif chemokine ligand 1 (CXCL1), tumor necrosis factor-α (TNF-α), interleukin (IL)-6, interferon-γ (IFN-γ), and IL-10. ELISA was used to measure the contents of acetylcholine (ACh) and acetylcholinesterase (AChE) in colonic tissues, and quantitative real-time PCR was performed to detect the mRNA expression of α7 nicotinic acetylcholine receptor (α7nAChR). In the second experiment, C57BL/6J mice were randomly divided into a normal group and a normal EA group, with 6 mice per group. Mice in the normal EA group received EA intervention on days 1, 3, 5 and 7. Immunofluorescence staining was applied to observe the activating effect of EA at ST37 on choline acetyltransferase (ChAT)-positive neurons in pelvic ganglia. In the third experiment, four ChAT-Ai14 reporter mice were used. Immunofluorescence staining was performed to observe the anatomical association between ChAT-positive nerve fibers and α7nAChR-positive cells in the colon. In the fourth experiment, 9 SD rats were collected, and neuroelectrophysiology was used to observe the influence of acupuncture at ST37 on the discharge activity of pelvic nerves.ResultsCompared with the normal group, the model group exhibited significantly reduced body weight (P<0.05, P<0.01), markedly shortened colon length (P<0.01), and elevated pathological scores of colonic tissues (P<0.001). The levels of pro-inflammatory factors including CXCL1, TNF-α, IL-6 and IFN-γ were increased (P<0.01, P<0.05), whereas the content of AChE and the mRNA expression of α7nAChR were significantly decreased (P<0.01). Compared with the model group, the EA group showed a markedly alleviated loss of body weight (P<0.05), recovered colon length (P<0.05), attenuated colonic tissue injury and decreased pathological scores (P<0.001). The colonic levels of pro-inflammatory factors CXCL1, TNF-α, IL-6 and IFN-γ were downregulated (P<0.05, P<0.01), while the anti-inflammatory factor IL-10 was upregulated (P<0.01). In addition, the AChE content and α7nAChR mRNA expression were significantly elevated in the EA group (P<0.01). EA at ST37 significantly activated ChAT-positive neurons in pelvic ganglia of colitic mice relative to the normal group (P<0.01). Anatomical proximity was observed between ChAT-positive nerve fibers and α7nAChR-positive cells in the colon of ChAT-Ai14 mice. Acupuncture at ST37 markedly increased the discharge frequency of pelvic nerves (P<0.001).ConclusionEA at ST37 alleviates colonic inflammatory response in mice by activating the sacral parasympathetic cholinergic neuroimmune pathway. The underlying mechanism may be related to EA-evoked activation of ChAT neurons in pelvic ganglia and pelvic nerves, which further regulates the anti-inflammatory effects mediated by colonic α7nAChR.
摘要:On the occasion of the 50th anniversary of the founding of Acupuncture Research, this paper systematically reviews the milestone value of the journal in promoting the scientification, standardization, and internationalization of acupuncture-moxibustion medicine. Integrating the core research achievements of the authors’ team over the past sixty-odd years—particularly the classic literature published in Acupuncture Research over the last half-century—this article deeply analyzes the development characteristics of modern acupuncture research in its transition from “empirical verification” to “mechanism-driven” science. Furthermore, it elucidates four core pathways for the high-quality development of the acupuncture research field: first, emphasizing the neurochemical principles of acupuncture analgesia to drive theoretical and technological innovation through heritage preservation; second, harnessing translational medicine as a developmental catalyst to integrate basic medicine and clinical medicine; third, spearheading the evolution of the “Clinical+X” paradigm via brain-computer interfaces; and fourth, prioritizing the standardization and internationalization of acupuncture research, while strengthening the building of multidisciplinary talent teams to enhance China’s voice and leadership in the field of global acupuncture governance. Ultimately, this work provides a robust theoretical foundation and forward-looking horizons for constructing a modern acupuncture medical system with Chinese characteristics and advancing the cause of global health.
关键词:Acupuncture research;Translational medicine;Brain-computer interface;Standardization;Talent cultivation;Clinical + X
摘要:This paper systematically elaborates the theories of stress, biological resilience and hormesis. Mild low-dose stress induces organismic overcompensation via preconditioning, activates nuclear factor erythroid 2-related factor 2 (Nrf2)-mediated antioxidant, autophagy, DNA repair and other pathways, and presents an inverted U-shaped biphasic dose-response relationship. Preconditioning is categorized into local and remote types, which exerts long-term protective effects on systemic organs via metabolic reprogramming as well as neurohumoral and immune pathways, with two distinct protective time windows (early and delayed). Numerous clinical trials have validated that remote ischemic preconditioning improves the prognosis of organ transplantation, stroke, hypertension and other diseases, yet its clinical application is limited by the requirement to predict injuries in advance. It proposes that somatic stimulation therapies including acupuncture, scraping and cupping act as mild stress-like interventions that fully conform to the six core characteristics of hormesis. By regulating oxidative stress, inflammatory response, mitochondrial function and DNA repair pathways, these therapies elevate systemic biological resilience, consistent with the evolutionary law that “mild stress confers benefits” and the traditional Chinese medicine health-preserving philosophy of tempering vitality through hardship as recorded in ancient texts. Acupuncture can be applied to perioperative care, chronic disorders, neurological diseases and full-life-cycle health management. Nevertheless, current research on acupuncture modulating resilience-related pathways remains insufficient, and in-depth exploration of its underlying basic mechanisms and clinical translation is still warranted.
Li Jiquan, Ma Tieming, Wang Wei, Chen Yiran, Zhao Yan, You Linxin, Zhang Jingyu, Wang Ying, Lin Xingxing, Zheng Qu, Zhao Yeyu, Li Sijia
DOI:10.13702/j.1000-0607.20260134
摘要:ObjectiveTo observe the antipruritic effect of electroacupuncture (EA) at antipruritic acupoints “Quchi” (LI11) and “Xuehai” (SP10) on skin itch in urticaria (UR) rats, so as to analyze the mechanism underlying the antipruritic effect of EA based on transient receptor potential vanilloid subtype 1 (TRPV1) channel-related proteins. Methods Anti-ovalbumin (OVA) serum was prepared by combined injection of the mixture of OVA and aluminum hydroxide immune adjuvant in 3 male SD rats. A total of 16 male SD rats were adopted to establish UR models via passive cutaneous anaphylaxis (PCA): diluted anti-OVA serum was intradermally injected into the rat back, followed by antigen challenge via tail vein injection of diluted OVA and Evans blue solution 48 h later, and model evaluation was conducted 0.5 h after challenge. Successfully modeled rats were randomly divided into a model group and an EA group, with 8 rats in each group; another 8 rats were set as the normal group. Starting from the 2nd day after model establishment, rats in the EA group received EA stimulation at bilateral LI11 and SP10, once daily for 5 consecutive days. The scratching frequency of rats on sensitized back skin before and after EA intervention was recorded, and the area of blue spots on sensitized back skin was measured. Sensitized back skin tissues were harvested for subsequent detections: colorimetric method was used to quantify Evans blue exudation in blue spots of sensitized skin; hematoxylin-eosin (HE) staining was performed to observe skin pathological lesions; transmission electron microscopy was applied to examine the ultrastructure of skin mast cells (MCs); toluidine blue staining was adopted to observe MC morphology and calculate the MC degranulation rate; ELISA was used to detect serum levels of immunoglobulin E (IgE), histamine (HIS), 5-hydroxytryptamine (5-HT), interleukin (IL)-1β, IL-5 and IL-6; immunohistochemistry was conducted to count phosphatidylinositol 4,5-bisphosphate (PIP2)-positive cells in skin tissues; Western blot was utilized to detect the protein expressions of protein kinase C (PKC), phosphorylated PKC (p-PKC) and TRPV1 in skin tissues. Results Compared with the normal group, the model group exhibited significantly increased scratching frequency, enlarged area of blue spots on sensitized skin and elevated Evans blue exudation (P<0.01), accompanied by severe inflammatory infiltration in sensitized skin. MCs showed increased basophilic secretory granules in cytoplasm, damaged granule membranes and decreased internal granule density, with a markedly elevated MC degranulation rate (P<0.01). Serum levels of IgE, HIS, 5-HT, IL-1β, IL-5 and IL-6 were significantly up-regulated (P<0.01). In addition, the number of PIP2-positive cells, protein expressions of PKC and TRPV1, as well as the p-PKC/PKC ratio in skin tissues were all increased (P<0.01). Compared with the model group, the EA group displayed reduced scratching frequency, smaller blue spot area and lower Evans blue exudation (P<0.05, P<0.01). Skin inflammatory lesions were alleviated; MCs presented fewer cytoplasmic basophilic secretory granules, restored intact granule membranes and higher internal granule density, alongside a decreased MC degranulation rate (P<0.01). Serum contents of IgE, HIS, 5-HT, IL-1β, IL-5 and IL-6 were notably decreased (P<0.01). Meanwhile, the number of PIP2-positive cells, expressions of PKC and TRPV1, and the p-PKC/PKC ratio in skin tissues were down-regulated (P<0.01, P<0.05).ConclusionEA at antipruritic acupoints LI11 and SP10 can relieve skin itch and wheal lesions in UR rats. Its therapeutic mechanism may be associated with inhibiting TRPV1 channel-related proteins in skin tissues and suppressing mast cell degranulation, thereby mitigating local inflammatory responses in lesioned skin.
Li Wei, Li Peifang, Zhang Daozong, Liu Hui, Liu Yan, Gao Zhi, Liu Xiaomeng
DOI:10.13702/j.1000-0607.20260294
摘要:ObjectiveTo investigate the ameliorative effect of electroacupuncture (EA) on cognitive dysfunction in Alzheimer’s disease (AD) mice, and observe whether EA exerts neuroprotective effects by regulating ubiquitin-proteasome system (UPS)-mediated microglial inflammatory response and the signal transducer and activator of transcription 3 (STAT3)/inflammasome/synaptic damage axis, so as to explore its underlying mechanism.MethodsWild-type mice were set as the normal control group. Tau-P301S transgenic mice were randomly divided into model group, EA group, sham EA group, EA combined with MG132 group and MG132 group, with 6 mice in each group. Mice in the EA group and EA+MG132 group received acupuncture at “Dazhui” (GV14), “Mingmen” (GV4), “Baihui” (GV20) and “Shenting” (GV24); electrodes were connected to GV20 and GV24 with dense-disperse waves at frequencies of 2 Hz/100 Hz for 20 min per session, once daily. A treatment course consisted of 6 consecutive days of acupuncture followed by 1 day of rest, with a total of 4 courses. Mice in the EA+MG132 group and MG132 group were injected with 2 μL MG132 (5 mmol/L) into the bilateral hippocampal CA1 region. Morris water maze test and open field test were performed to evaluate cognitive function and anxiety-like behaviors. Hematoxylin-eosin (HE) staining was used to observe pathological changes in hippocampal tissues. Transmission electron microscopy (TEM) was adopted to detect ultrastructural damage of hippocampal neuronal synapses and mitochondria. Immunofluorescence staining was applied to examine microglial activation and the fluorescence positive expression of transmembrane protein 16F (TMEM16F) in the hippocampus. Western blot was utilized to detect the expression levels of UPS-related proteins, STAT3/suppressor of cytokine signaling 3 (SOCS3) pathway proteins, inflammasome components, inflammatory factors and synaptic proteins.ResultsCompared with the normal control group, the model group exhibited significantly prolonged escape latency (P<0.05, P<0.01), markedly reduced times of crossing the original platform (P<0.05) and shortened residence time in the target quadrant (P<0.05). In the open field test, total movement distance, times of entering the central zone, residence time and movement distance in the central zone were all significantly decreased (P<0.05). In the hippocampal CA1 region of model mice, neurons were disorganized with shrunken somata, severe pyknosis, widened intercellular spaces and reduced neuronal number; mitochondria were swollen with fractured or disappeared cristae and cytoplasmic vacuolation; synaptic clefts were widened and synaptic vesicles were decreased. The fluorescence intensity and number of ionized calcium-binding adapter molecule 1 (Iba1)-positive cells, as well as the number of TMEM16F/Iba1 double-positive cells in the hippocampus were significantly elevated (P<0.01). The protein expression ratios of phosphorylated signal transducer and activator of transcription 3 to total STAT3 (p-STAT3/STAT3), NOD-like receptor pyrin domain-containing protein 3 (NLRP3), cysteinyl aspartate specific proteinase-1 (Caspase-1), tumor necrosis factor-α (TNF-α) and cyclooxygenase 2 (COX-2) in hippocampal tissues were remarkably up-regulated (P<0.01). Meanwhile, the protein levels of SOCS3, ubiquitin carboxyl-terminal hydrolase L1 (UCHL1), interleukin-10 (IL-10), postsynaptic density protein 95 (PSD95) and synaptophysin (SYP) were markedly down-regulated (P<0.01). Compared with the model group, all above indicators were significantly improved in the EA group (P<0.01); hippocampal CA1 neurons were arranged more regularly with relatively intact somata, alleviated pyknosis and narrowed intercellular spaces; mitochondrial swelling was relieved with relatively complete cristae, cytoplasmic vacuolation was reduced, and partial recovery of synaptic structure was observed. In contrast with the EA group, almost all indicators were reversed in the sham EA group and EA+MG132 group, except for SYP expression in the sham EA group (P<0.05).ConclusionEA can alleviate cognitive dysfunction in AD model mice. Its therapeutic effect may partially depend on the regulation of UPS dysfunction-related microglial inflammation, which is associated with the normalization of UPS-mediated abnormal STAT3/SOCS3 signaling, suppression of inflammasome activation and alleviation of synaptic damage.
关键词:Electroacupuncture;Alzheimer’s disease;Microglia;Inflammasome;Synaptic damage;Ubiquitin-proteasome system
Luo Qiuping, Li Hongying, Yang Xinyu, Lu Chunxia, Cui Jin
DOI:10.13702/j.1000-0607.20251424
摘要:ObjectiveTo observe the effect of electroacupuncture (EA) on ovarian function in perimenopausal rats with ovarian function decline, and to explore its potential molecular mechanism underling delaying ovarian aging through the Kisspeptin (KP)/follicle-stimulating hormone receptor (FSHR) pathway.MethodsForty female SD rats were included in the present study. Ten rats with regular estrous cycles were selected as the control group. Thirty perimenopausal (disordered estrous cycle) rats with ovarian function decline were randomly divided into model, EA, and EA plus antagonist (KP234, an antagonist for KP) groups, with 10 rats in each group. For rats of the EA and EA+antagonist groups, EA (0.1—1 mA, 2 Hz) was applied to Guanyuan (CV4), Qihai (CV6), and bilateral Taichong (LR3) and Taixi (KI3) for 20 min, once daily for 14 consecutive days. After EA intervention, rats of the EA+antagonist group received intraperitoneal injection of KP antagonist KP234 (100 nmol/kg) once every other day for 14 days. During the intervention, vaginal smears were performed daily, and rapid Gram staining was conducted to determine the estrous cycles. Twenty-four hours after the last intervention, blood samples and ovarian tissues were collected for measuring the contents of serum anti-Müllerian hormone (AMH), follicle-stimulating hormone (FSH), luteinizing hormone (LH) and estradiol (E2) with ELISA. The ovarian histomorphology and follicular development were observed by H.E. staining. The reactive oxygen species (ROS) levels in the ovarian tissue were measured by immunofluorescence assay. The mRNA expression levels of Sirtuin 1 (Sirt1), tumor suppressor protein (p53) and cyclin-dependent kinase inhibitor 1 (p21) in the ovarian tissue were detected by real-time quantitative PCR. The immunoactivity of KP, KP receptor (Kiss1R) and FSHR, and the KP/Kiss1R colocalization in the ovarian tissue were detected by immunofluorescence staining. The protein expression levels of ovarian Sirt1, p53, p21, KP, Kiss1R and FSHR were detected by western blot.ResultsAfter modeling and treatment, all the rats (100%) of the model group, 20% in the EA group and 80% in the EA+inhibitor group had a disordered estrous cycle, being significantly lower in the EA group than in the model group (P<0.001), and significantly higher in the EA+inhibitor group than in the EA group (P<0.05). Compared with the control group, the model group showed a considerable decrease in the contents of serum AMH and E2, number of growing follicles, expression level of Sirt1 mRNA, and immunoactivity of Kiss1R and FSHR, and percent of colocalized area of KP/KissR, as well as the expression levels of Sirt1, Kiss1R and FSHR proteins (P<0.001, P<0.01), and a notable increase in the contents of serum FSH and LH, number of atretic follicles, ovarian ROS level, mRNA expression levels of p53 and p21, immunoactivity of KP, and protein expression of p53, p21 and KP (P<0.05, P<0.01, P<0.001). Following the intervention, modeling induced decrease and increase of all the indexes mentioned above were reversed by EA (P<0.001, P<0.05, P<0.01) rather than by EA + inhibitor, suggesting an elimination of the therapeutic effect after administration of KP234, an antagonist for KP. Additionally, H.E. staining of the ovarian tissue displayed that in the model group, early on, the follicular collapse could be seen, with disordered arrangement of granulosa cells and theca cells, while in the later stage, irregularly shaped annular zona pellucida could be seen. The arrangement of granular cells in the EA group became more regular.ConclusionEA can improve the ovarian reserve function and delay ovarian aging by attenuating local oxidative stress and cellular senescence in the ovary through regulating the KP/FSHR pathway.
关键词:Electroacupuncture;Perimenopausal ovarian function decline;Ovarian aging;Oxidative stress;KP/FSHR signaling pathway
Wang Yujun, Lin Wei, Chen Jiaqi, Zhang Yihang, Chen Yang, Guo Jing, Yin Tao, Zeng Fang
DOI:10.13702/j.1000-0607.20251242
摘要:ObjectiveTo analyze the symptom clusters, acupoint selection rules, symptom-acupoint association rules, and acupoint combinations for functional gastrointestinal disorders (FGIDs) in ancient literature based on data mining technology.MethodsRelevant texts regarding acupuncture and moxibustion treatment of FGIDs were collected from 68 ancient acupuncture classics, such as Ling Shu (Miraculous Pivot) and Zhen Jiu Jia Yi Jing (Systematic Classic of Acupuncture and Moxibustion). According to inclusion and exclusion criteria, relevant information was extracted to establish a prescription database. Descriptive statistics were performed on the frequency, meridian tropism, and specific acupoints, and a map of high-frequency acupoints was drawn. Python 3.3.2 software was used to conduct symptom-acupoint association rule analysis and plot a complex network relationship diagram. Cluster analysis was performed on symptoms with a frequency >5. R 4.4.2 software was utilized to conduct heatmap, cluster, and co-occurrence network analyses on the top 20 high-frequency acupoints.ResultsA total of 566 texts were included, involving 145 symptoms and 161 acupoints. The top four acupoints in terms of application frequency were Zhongwan (CV12), Zusanli (ST36), Neiguan (PC6), and Pishu (BL20). The meridians involved were mainly the Conception Vessel (Ren Mai), Bladder Meridian of Foot-Taiyang, Stomach Meridian of Foot-Yangming, and Spleen Meridian of Foot-Taiyin. Among the specific acupoints, Five-Shu acupoints, Front-Mu acupoints, and Eight-Influential acupoints were frequently used among the specific acupoints. The symptom-acupoint association rules revealed a group of correlated symptoms: retching, vomiting, regurgitation, and indigestion, which embodied the acupoint selection rules corresponding to pathogenesis. The symptom cluster analysis yielded 8 categories, including stomach qi counterflow syndrome, food accumulation in the stomach and intestines syndrome, qi-stagnation syndrome, and stomach-yang deficiency syndrome. The acupoint cluster analysis resulted in 4 categories, including the group for supplementing qi, resolving dampness and removing blood stasis, and the group for unblocking the heart and diaphragm. Conclusion Acupuncture and moxibustion for treating FGIDs commonly utilizes acupoints such as Zhongwan (CV12), Zusanli (ST36), Neiguan (PC6), and Pishu (BL20). Certain symptom clusters are correlated with specific subtypes of modern FGIDs.
Liu Jiabao, Ren Hongxia, Lin Qishun, Zhao Shuai, Li Tong, Jian Hongwei, Sun Hong, Yu Lingling, Jing Xianghong, Li Man, Zhang Hongxing
DOI:10.13702/j.1000-0607.20260402
摘要:ObjectiveTo explore the mechanism by which electroacupuncture (EA) combined with antibiotics regulates the vagus-adrenal axis via the dorsal motor nucleus of vagus (DMV), promotes catecholamine secretion, attenuates inflammatory storm, and improves survival rate in septic mice.MethodsExperiment 1: Fifty C57BL/6 mice were randomly divided into sham operation group, model group, antibiotic group, EA group, and EA + antibiotic group, with 10 mice in each group. Cecal ligation and puncture (CLP) was performed to establish the sepsis model. Mice in the EA group received 15-min EA at bilateral “Zusanli” (ST36), once daily for 3 consecutive days. Mice in the antibiotic group were intraperitoneally injected with imipenem-cilastatin sodium (30 mg/kg) every 12 h for 3 consecutive days. The 3-day survival rate and body weight changes of mice in each group were recorded. ELISA was adopted to detect serum levels of interleukin-6 (IL-6) and tumor necrosis factor-α (TNF-α). HE staining was used to observe pathological changes and the number of inflammatory cells in cecal tissues. Whole-brain optical clearing was performed to detect the expression of proto-oncogene c-Fos protein in the DMV region. Experiment 2: Fifty Chat-Cre mice were randomly assigned to sham operation + empty virus group, model + empty virus group, EA-antibiotic combination + inhibitory virus group, EA-antibiotic combination + empty virus group, and antibiotic + activating virus group, with 10 mice per group. Corresponding viruses were injected into the DMV 3 weeks before model establishment. CLP operation was conducted 3 weeks after virus injection, followed by daily intraperitoneal injection of clozapine N-oxide (CNO, 3 mg/kg), combined with antibiotics or EA treatment (the same interventions and parameters as Experiment 1. Serum IL-6 and TNF-α concentrations were measured by ELISA; cecal inflammatory infiltration was evaluated via HE staining. Ultra-high performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) was applied to detect serum catecholamines including dopamine (DA), epinephrine (E), norepinephrine (NE), as well as their metabolites 3-methoxytyramine (3-MT), normetanephrine (NMN), and metanephrine (MN) levels. Immunofluorescence staining was used to quantify the counts of programmed death receptor-1 positive (PD-1+) cells and forkhead box protein P3 positive / cluster of differentiation 4 positive (FOXP3+/CD4+) cells in cecal tissues.ResultsExperiment 1: Compared with the sham operation group, the model group exhibited markedly decreased 3-day survival rate and body weight (P<0.000 1), elevated serum IL-6 and TNF-α levels (P<0.000 1), reduced concentrations of DA, E, NE and their metabolites 3-MT, NMN, MN (P<0.01, P<0.000 1), and increased inflammatory cell infiltration in cecum (P<0.000 1). Compared with the model group, the EA + antibiotic group showed improved survival rate (P<0.05), alleviated body weight loss (P<0.000 1), decreased serum IL-6 and TNF-α levels (P<0.000 1), reduced inflammatory cell infiltration (P<0.000 1), and elevated c-Fos density in DMV (P<0.000 1). Moreover, all above indicators were superior to those in the single EA or single antibiotic group to varying degrees. Experiment 2: In comparison with the sham operation + empty virus group, the model + empty virus group had higher serum inflammatory factor levels and more cecal inflammatory infiltrates (P<0.0001), and lower levels of catecholamines and their metabolites, as well as fewer PD-1+ and FOXP3+/CD4+ cells in cecal tissues (P<0.000 1). Compared with the model + empty virus group, the EA-antibiotic combination + empty virus group, EA-antibiotic combination + inhibitory virus group, and antibiotic + activating virus group displayed reduced serum inflammatory factors and relieved cecal inflammatory infiltration (P<0.000 1, P<0.01, P<0.05), accompanied by increased levels of catecholamines and metabolites, and more PD-1+ and FOXP3+/CD4+ cells in cecum (P<0.001, P<0.05, P<0.01, P<0.0001). Relative to the EA-antibiotic combination + empty virus group, all the above indicators were reversed in the EA-antibiotic combination + inhibitory virus group (P<0.01, P<0.001, P<0.05).ConclusionEA at ST36 combined with antibiotics exerts optimal therapeutic effects on CLP-induced septic mice. The underlying mechanism may involve activating cholinergic neurons in DMV, regulating the vagus-adrenal axis to facilitate catecholamine secretion, suppressing excessive inflammatory response, and consequently elevating survival rate.
Liang Tingting, Zhao Yu, Wang Tianlang, Zhang Jiahao, Wang Xihan, Wang Xining, Feng Yifan, Guo Yue, Liu Yunxiao, Qu Yi, Xue Yanan, Liu Jidong
DOI:10.13702/j.1000-0607.20260535
摘要:ObjectiveTo clarify the pathway by which electroacupuncture at “Taichong” (LR3) and “Zusanli” (ST36) alleviates low-grade duodenal inflammation in functional dyspepsia (FD) rats via regulating the Hippo/Yes-associated protein (YAP) signaling pathway.MethodsA total of 21 male SD rats (SPF grade) were enrolled and randomly divided into the blank group, the model group, and the electroacupuncture (EA) group (n = 7). The FD model was established through a combined induction strategy involving simultaneous administration of exhaustive swimming, tail-clamping stimulation, and food restriction/overeating protocols, with continuous intervention lasting 21 days. Rats in the EA group received EA stimulation at LR3 and ST36, once daily for 20 min, 6 consecutive days followed by 1-day rest each course , with 2 courses administered in total. The general physical conditions and body weight of rats in each group were monitored throughout model establishment and treatment. Upon completion of intervention, the 3-h food intake, gastric emptying rate and small intestinal transit rate of rats were measured. HE staining was performed to observe pathological changes in duodenal mucosa. ELISA was adopted to detect serum levels of motilin (MLT) and gastrin (GAS). Western blot was used to quantify the protein expressions of YAP, transcriptional coactivator with PDZ-binding motif (TAZ), phosphorylated (p)-YAP and p-TAZ in duodenal tissues. Immunohistochemistry was applied to determine the positive expression of interleukin (IL)-1β, IL-6 and tumor necrosis factor-α (TNF-α) in duodenum. Immunofluorescence staining was conducted to detect the positive expression of Occludin and zonula occludens-1 (ZO-1) in duodenal tissues. Quantitative real-time PCR was utilized to test the mRNA expression levels of YAP, TAZ, B-cell lymphoma-2 (Bcl-2) and Bcl-2-associated X protein (Bax) in duodenum. TUNEL staining was carried out to assess the apoptotic rate of duodenal epithelial cells.ResultsCompared with the blank group, rats in the model group exhibited severe lethargy, manifested as narrowed eyes, curled recumbency, dry and messy fur, and loose stools; the body weight, 3-h food intake, gastric emptying rate and small intestinal transit rate were markedly decreased (P<0.05, P<0.01); massive inflammatory infiltration was observed in the lamina propria of duodenal tissues; serum MLT and GAS concentrations were significantly reduced (P<0.01); the protein expression ratios of p-YAP/YAP and p-TAZ/TAZ in duodenum were notably elevated (P<0.01); the positive expression of IL-1β, IL-6 and TNF-α in duodenal tissues was upregulated (P<0.01). The mean fluorescence intensity of Occludin and ZO-1 in duodenum was remarkably lowered (P<0.01). The mRNA level of Bax in duodenal tissues was significantly increased (P<0.01), while the mRNA expressions of YAP, TAZ and Bcl-2 were distinctly downregulated (P<0.01). The apoptotic rate of intestinal epithelial cells was markedly higher (P<0.01). After EA intervention, relative to the model group, rats in the EA group showed improved mental state, enhanced locomotor activity, glossy fur and formed stools, the inflammatory infiltration in the duodenal lamina propria was alleviated, and all the indicators above were reversed (P<0.01, P<0.05).ConclusionElectroacupuncture at LR3 and ST36 can effectively relieve symptoms of FD, improve the general physical status and suppress low-grade duodenal inflammation in FD rats. Its therapeutic mechanism may be associated with the Hippo/YAP signaling pathway.
LI Hong-ying, LUO Hong-yan, ZHU Min, XUE Kai-yang, LU Xiao-xiao, CUI Jin
当前状态:二校优先
DOI:10.13702/j.1000-0607.20250861
摘要:ObjectiveTo investigate the effects of herbal cake-separated moxibustion combined with atorvastatin on lipid metabolism, aortic pathological structure, plaque deposition, and integrin/ yes-associated protein (YAP)/ c-Jun N-terminal kinase (JNK) signaling pathway in ApoE-/- atherosclerosis (AS) mice, and to explore its preventive and therapeutic mechanisms for AS.MethodsMale C57BL/6J mice were used as the blank group, and male ApoE-/- mice were randomly divided into the model group, medicine group, herbal cake-separated moxibustion group, and combined treatment group, with 9 mice per group. High-fat diet was used to establish the AS model. The herbal cake-separated moxibustion group received moxibustion at “Danzhong” (CV17) and “Shenque” (CV8) acupoints, once every other day, 3 times a week. The medicine group received atorvastatin calcium tablet via gavage, 3 mg·kg-1·d-1, once daily. The combined treatment group received an integrated approach that combined the interventions of the medicine group and herbal cake-separated moxibustion group. All groups were treated for 8 weeks. Body weight were observed before and after treatment. HE staining was used to observe aortic pathological morphology, and Oil Red O staining to observe aortic lipid plaque area. Biochemical analysis was used to detect serum triglyceride (TG), total cholesterol (TC), low-density lipoprotein cholesterol (LDL-C), and high-density lipoprotein cholesterol (HDL-C) levels. Serum vascular endothelial growth factor (VEGF), endothelin-1 (ET-1), interleukin (IL)-6, IL-8, and tumor necrosis factor-α (TNF-α) levels were detected by ELISA, and serum nitric oxide (NO) level was detected by colorimetric assay. Western blot was used to detect protein expression levels of integrin αVβ3,YAP, phosphorylated (p)-YAP, p-JNK1/2, intercellular adhesion molecule-1 (ICAM-1), and vascular cell adhesion molecule-1 (VCAM-1) in the aorta.ResultCompared to the blank group, the model group exhibited increased body weight (P<0.01); thickened aortic intima, foam cells and lipid deposition, hypertrophic edema of smooth muscle cells, thinning of elastic fibers, widened endothelial cell gaps, and rupture of elastic membranes; significantly increased arterial lipid plaque area (P<0.01); elevated serum TC, TG, LDL-C, IL-6, IL-8, TNF-α, ET-1, and VEGF levels (P<0.01), while HDL-C and NO levels decreased (P<0.01); increased protein expression of integrin αVβ3, YAP, p-JNK1/2, ICAM-1, and VCAM-1 (P<0.01), and reduced p-YAP protein expression (P<0.01) in the aorta. Compared to the model group, the medicine group, herbal cake-separated moxibustion group, and combined treatment group showed reduced body weight (P<0.01), with a significantly lower weight change difference than that of the model group (P<0.01); more regular aortic lumen structure, thinner intima, reduced foam cells and lipid deposition, alleviated smooth muscle cell edema, hypertrophy, and inflammatory infiltration; all other indicators also showed significant improvement compared to the model group (P<0.01, P<0.05). Compared to the herbal cake-separated moxibustion group, the medicine group and combined treatment group exhibited reduced plaque area (P<0.05, P<0.01), increased serum NO level (P<0.01), decreased LDL-C, ET-1, and VEGF levels (P<0.01), and reduced protein expressions of p-JNK1/2, ICAM-1, and VCAM-1 in the aorta (P<0.05, P<0.01); the medicine group also showed decreased serum TC, TG, IL-6, IL-8, and TNF-α levels (P<0.01), increased HDL-C level (P<0.01), reduced integrin αVβ3 and YAP protein expressions in the aorta (P<0.01, P<0.05), and increased p-YAP protein expression (P<0.05). Compared to the medicine group and the herbal cake-separated moxibustion group, the combined treatment group exhibited a greater reduction in body weight (P<0.05, P<0.01), decreased serum levels of TC, TG, IL-6, IL-8, and TNF-α, as well as reduced expressions of integrin αVβ3 and YAP proteins in the aorta (P<0.01, P<0.05), elevated serum HDL-C level (P<0.01), and increased p-YAP protein expression (P<0.01).ConclusionHerbal cake-separated moxibustion combined with atorvastatin exerts synergistic effects, ameliorates serum lipid profiles, mitigates endothelial dysfunction of aorta, and attenuates inflammatory responses in AS mice, potentially via modulation of the integrin/YAP/JNK signaling pathway in the aorta.
SHI Rong, MA Tie-ming, SUN Shi-jing, JIANG Jing, LI Jia-xin, WU Qiong, ZHOU Zi-xi, ZHANG Jia-wei, YANG Rui-rui, ZHAO Xi-tong, MA Jun-jie, ZHANG Xiao-qing, CHEN Yi-ran
当前状态:二校优先
DOI:10.13702/j.1000-0607.20251014
摘要:ObjectiveTo observe the effect of “Tiaoshen Tongluo”(mind regulation and meridian-collateral dredging) electroacupuncture (EA) on the nuclear factor erythroid 2-related factor 2 (Nrf2)/triggering receptor expressed on myeloid cells 2 (TREM2) signaling pathway and hippocampal microglial polarization in rats with learning-memory impairment induced by middle cerebral artery occlusion/reperfusion (MCAO/R).MethodsMale SD rats were randomly divided into 7 d sham group (n=10), 14 d sham group (n=10), and model group. The MCAO/R model was established by Koizumi intraluminal filament MCAO method. Rats meeting the criteria of learning-memory impairment due to MCAO/R,identified by the Morris water maze tests, were randomly assigned to 7 d model, 7 d EA, 14 d model and 14 d EA groups, with 10 rats in each group. Rats of the EA groups received “Tiaoshen Tongluo” EA stimulation at “Baihui” (GV20), “Yintang” (GV24+), and bilateral “Taichong” (LR3) and “Hegu” (LI4) for 10 min per session, once daily, for 7 or 14 consecutive days. The modified neurological severity score (mNSS) was used to assess the neurological deficits before and after the intervention. Morris water maze test was conducted to evaluate learning and memory abilities. HE staining was used to observe histopathological changes, and Nissl staining was applied to examine neuronal morphology and Nissl bodies in the hippocampal CA1 region. Immunofluorescence double staining was employed to determine the proportions of CD68+/Iba1+ (M1 type) and CD206+/Iba1+ (M2 type) positive cells in the hippocampus tissue. Western blot was used to measure the relative protein expression levels of Nrf2, TREM2, DNAX activator protein 12(DAP12), tumor necrosis factor-α(TNF-α) and transforming growth factor-β(TGF-β) in the hippocampus tissue.ResultsCompared with the 7 d and 14 d sham groups, the 7 d and 14 d model groups had a significant increase in the mNSS (P<0.01), and an obvious down-regulation in expression levels of hippocampal Nrf2, TREM2, and DAP12 proteins (P<0.05, P<0.01), and loose arrangement of neurons and reduction in the number of Nissl bodies in the hippocampal CA1 region. In contrast to the 14 d sham group, the 14 d model group had a notable increase in the escape latency, the number of CD68+/Iba1+ dual labeled cells, and the expression level of TNF-α in the hippocampus (P<0.05, P<0.01), and a striking decrease in the number of original platform crossings and time spent in the target quadrant (P<0.01). Compared with the 7 d and 14 d model groups, the 7 d and 14 d EA groups exhibited reduction of mNSS and TNF-α expressions (P<0.01, P<0.05), up-regulation of expression levels of Nrf2 and TREM2 proteins (P<0.05, P<0.01), alleviated arrangement of CA1 neurons, and increased Nissl bodies. Compared with the 14 d model group, the 14 d EA group showed a decrease in the escape latency and the number of CD68+/Iba1+ dual labeled cells (P<0.01), and an increase in the number of original platform crossings and time spent in the target quadrant, number of CD206+/Iba1+ dual labeled cells, and the expression of DAP12 and TGF-β proteins(P<0.05, P<0.01). Compared with the 7 d EA group, the 14 d EA group showed a further reduction in mNSS score (P<0.05), and a further up-regulation of expression of TREM2 protein (P<0.05), a more compact neuronal arrangement in the CA1 region with the morphological structure being close to the normal.Conclusion“Tiaoshen Tongluo” EA can improve learning-memory ability in MCAO/R rats, which may be associated with its functions in activating the Nrf2/TREM2 signaling pathway, inhibiting microglial M1 polarization and promoting M2 polarization. In addition, the therapeutic effect of 14 d EA is better than that of 7 d EA.
WANG Shi-lin, HE Long, YANG Jing, ZHANG Na, SUN Yuan-zheng, LIU Guan-ping
当前状态:二校优先
DOI:10.13702/j.1000-0607.20251237
摘要:ObjectiveTo investigate the regulatory effect of “Jiaji” (EX-B2) electroacupuncture (EA) on the NOD-like receptor thermal protein domain-associated protein 3 (NLRP3) inflammasome in the lumbar spinal cord and to explore its mechanism in improving limb dysfunction in amyotrophic lateral sclerosis (ALS) mice.MethodsFifty-four mice carrying the human superoxide dismutase 1 G93A (hSOD1G93A) gene were randomly divided into model, Jiaji EA, and inhibitor groups, with 18 mice per group. Eighteen mice not carrying the hSOD1G93A gene served as wild group. At 60 d of age, the Jiaji EA group was given EA at L1—L2 and L5—L6 EX-B2 points with continuous wave, 1 mA, 2 Hz. The needles retained for 20 min, twice every week. The inhibitor group received intraperitoneal injection of the NLRP3 inhibitor (10 mg/kg, twice every week). All treatments were administered for 4 consecutive weeks. The survival period of mice in each group was observed to assess disease progression; the rotarod test was conducted to evaluate motor coordination and limb motor function; HE staining was used to observe pathological changes in the gastrocnemius, and muscle atrophy was assessed by measuring the cross-sectional area of the gastrocnemius fiber; Nissl staining was performed to evaluate the number of motor neurons in the lumbar spinal cord anterior horns; the protein expressions of NLRP3, Caspase-1, apoptosis-associated speck-like protein (ASC), interleukin-1β (IL-1β), interleukin-18 (IL-18), and tumor necrosis factor-α (TNF-α) in the lumbar spinal cord were detected by Western blot; the mRNA expressions of NLRP3, Caspase-1, ASC, IL-1β, IL-18, and TNF-α were detected by real-time quantitative PCR.ResultsCompared with the wild group, mice in the model group exhibited shortened survival time (P<0.01); reduced time spent on the rotating rod starting from the 13th week of age (P<0.05); significantly shorter and rounded gastrocnemius fibers with enlarged inter-fiber spaces, nuclear translocation and leakage. The cross-sectional area of the gastrocnemius fibers was reduced (P<0.01); the number of motor neurons in the spinal anterior horns was decreased (P<0.01); protein and mRNA expressions of NLRP3, Caspase-1, ASC, IL-1β, IL-18, and TNF-α were increased in the lumbar spinal cord (P<0.01). Compared with the model group, survival time was prolonged in the Jiaji EA and inhibitor groups (P<0.01); rotarod time was prolonged starting from the 13th week of age (P<0.05); gastrocnemius fibers exhibited more regular margins and larger morphology, with reduced inter-fiber spaces, and reduced number of rounded fibers and improvement in nuclear translocation and leakage; cross-sectional areas of gastrocnemius fiber were increased (P<0.01); the numbers of motor neurons in the spinal anterior horns were increased (P<0.01); protein and mRNA expressions of NLRP3, Caspase-1, ASC, IL-1β, IL-18, and TNF-α decreased in the lumbar spinal cord (P<0.05, P<0.01). Compared with the inhibitor group, mice in the Jiaji EA group exhibited prolonged rotarod time from the 17th week of age (P<0.05); the cross-sectional area of the gastrocnemius fiber increased (P<0.05); protein and mRNA expressions of NLRP3, Caspase-1 and IL-18 in the lumbar spinal cord were higher (P<0.05, P<0.01), while protein and mRNA expressions of TNF-α were lower (P<0.05, P<0.01).ConclusionEX-B2 EA improves limb function and prolongs survival in ALS mice, with its underlying mechanism potentially involving the alleviation of neuroinflammation through inhibition of NLRP3 inflammasome activation.