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1.成都中医药大学针灸推拿学院,成都610075
2.成都中医药大学养生康复学院,成都610075
3.针灸防治老年疾病教育部重点实验室,成都610075
姚俊鹏,Email: yjpcdtcm@163.com
收稿:2025-08-19,
修回:2025-09-09,
网络首发:2026-04-28,
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王雯,李艳秋,曾伟健,等.电针对功能性便秘小鼠结肠线粒体未折叠蛋白反应相关蛋白的影响[J].针刺研究,XXXX,XX(XX):1-9.
WANG Wen,LI Yan-qiu,ZENG Wei-jian,et al.Effect of electroacupuncture on mitochondrial unfolded protein response-associated proteins in the colon tissue of mice with functional constipation[J].Acupuncture Research,
王雯,李艳秋,曾伟健,等.电针对功能性便秘小鼠结肠线粒体未折叠蛋白反应相关蛋白的影响[J].针刺研究,XXXX,XX(XX):1-9. DOI: 10.13702/j.1000-0607.20250883.
WANG Wen,LI Yan-qiu,ZENG Wei-jian,et al.Effect of electroacupuncture on mitochondrial unfolded protein response-associated proteins in the colon tissue of mice with functional constipation[J].Acupuncture Research, DOI:10.13702/j.1000⁃0607.20250883.
目的
2
观察电针“天枢”“上巨虚”对功能性便秘(FC)模型小鼠结肠线粒体未折叠蛋白反应(UPR
mt
)相关蛋白表达的影响,探讨电针改善FC症状的潜在机制。
方法
2
18只雄性C57BL/6J小鼠随机分为空白组、模型组、电针组,每组6只。采用洛哌丁胺灌胃14 d构建FC小鼠模型。电针组采用电针交替干预单侧“天枢”和“上巨虚”(疏密波,3 Hz/15 Hz),每周连续治疗5次,共干预10次,空白组和模型组仅同等固定。采用首粒蓝便排出时间、6 h排便颗粒数、粪便性状评分、含水率和小肠推进率评估小鼠胃肠传输功能,HE染色观察结肠组织病理改变,透射电镜观察结肠组织线粒体超微结构,流式细胞术检测结肠组织线粒体膜电位和活性氧(mtROS)水平,比色法测定结肠组织线粒体ATP含量,免疫组织化学法检测UPR
mt
关键标志分子热休克蛋白-60(HSP60)和LON蛋白酶1(LONP1)在结肠组织的表达情况。
结果
2
与空白组相比,模型组小鼠首粒蓝便排出时间延长(
P
<
0.01),6 h内排便粒数减少(
P
<
0.01),粪便性状评分、含水率及小肠推进率降低(
P
<
0.01,
P
<
0.05),结肠组织线粒体明显肿胀,膜破损、嵴断裂严重,线粒体膜电位降低(
P
<
0.01),ATP含量下降(
P
<
0.01),mtROS水平升高(
P
<
0.01),结肠平滑肌细胞(SMC)中HSP60、LONP1蛋白阳性表达减少(
P
<
0.01)。干预后,与模型组比较,电针组首粒蓝便排出时间缩短(
P
<
0.01),6 h排便颗粒数增加(
P
<
0.01),粪便性状评分、含水率及小肠推进率升高(
P
<
0.01,
P
<
0.05),结肠组织线粒体轻度肿胀,膜、嵴损伤明显改善,线粒体膜电位升高(
P
<
0.01),ATP水平升高(
P
<
0.01),mtROS水平降低(
P
<
0.01),SMC中HSP60、LONP1阳性表达升高(
P
<
0.05)。
结论
2
电针“天枢”“上巨虚”能有效促进FC小鼠肠道传输功能,修复结肠SMC线粒体结构和功能,其机制可能与上调SMC中HSP60和LONP1的表达,进而激活UPR
mt
相关。
Objective
2
To observe the effect of electroacupuncture (EA) at “Tianshu” (ST26) and “Shangjuxu” (ST37) acupoints on the expression of mitochondrial unfolded protein response (UPR
mt)
- related proteins in the colon tissue of functional constipation (FC) model mice, so
as to explore its potential mechanisms underlying improvement of FC symptoms.
Methods
2
Eighteen male C57BL/6J mice were randomly divided into three groups: normal control (NC), FC model (M), and EA (
n
=6 per group). The FC model was established by gavage of Loperamide suspension (1 mg/mL, 0.1 mL/10 g), twice daily for 14 days. The EA group received alternating unilateral EA stimulation of ST26 and ST37(3 Hz/15 Hz, an electric current intensity being able to induce slight tremor of the mouse’s limbs) for 20 min, once daily, 5 days a week for 2 weeks. The mice in the NC and M groups underwent equivalent restraint. Gastrointestinal transit function was assessed by measuring the first blue stool discharge time, 6-hour fecal pellet count, stool character score, rate of water content, and intestinal propulsion rate. Histopathological changes of the colon tissue were examined by H.E. staining, and the mitochondrial ultrastructure of colonic smooth muscle cells (SMCs) was observed by using a transmission electron microscopy (TEM). The mitochondrial function was comprehensively evaluated by measuring the mitochondrial membrane potential (MMP) and the mitochondrial reactive oxygen species (mtROS) level was assayed using flow cytometry, and the mitochondrial ATP content in the colon tissue was measured using colorimetric assay. The immunohistochemistry stain was employed to detect the expression of key UPR
mt
markers heat shock protein-60 (HSP60) and LON protease 1 (LONP1) in the colon tissue.
Results
2
Compared with the NC group, the M group exhibited a significant increase in the first blue stool discharge time and average immunofluorescence intensity of mtROS (
P
<
0.01), and a considerable reduction in the 6-hour fecal pellet count, stool character score, rate of water content, small intestine propulsion rate, MMP, ATP content, and the expression of HSP60 and LONP1 proteins in the colon tissue (
P
<
0.0
5,
P
<
0.01). Following EA intervention, compared to the M group, the EA group showed a marked decrease in the first black stool discharge time and average immunofluorescence intensity of mtROS (
P
<
0.01), and a notable increase in the 6-hour fecal pellet count, stool character score, rate of water content, intestine propulsion rate, MMP, ATP content, and the expression of HSP60 and LONP1 proteins in the colon tissue (
P
<
0.05,
P
<
0.01). The results of morphological structure showed that the mitochondria in the colon tissue were significantly swollen, with membrane damage and severe cristae rupture in the model group, while in the EA group, the mitochondrial swelling was mild, with significant improvement in the membrane and cristae integrity of the SMCs.
Conclusion
2
EA at ST26 and ST37 can effectively ameliorate gastrointestinal dysfunction and restore mitochondrial integrity in colonic SMCs of FC mice, which may be associated with its functions in up-regulating the expression of HSP60 and LONP1 proteins to activate UPR
mt
.
Barberio B , Judge C , Savarino E V , et al . Global prevalence of functional constipation according to the Rome criteria: a systematic review and meta-analysis [J]. Lancet Gastroenterol Hepatol , 2021 , 6 ( 8 ): 638 - 648 .
Black C J , Drossman D A , Talley N J , et al . Functional gastrointestinal disorders: advances in understanding and management [J]. Lancet , 2020 , 396 ( 10263 ): 1664 - 1674 .
Hao W S , Cha R T , Wang M Z , et al . Ligand-modified gold nanoparticles as mitochondrial modulators: regulation of intestinal barrier and therapy for constipation [J]. ACS Nano , 2023 , 17 ( 14 ): 13377 - 13392 .
Reymond Sutandy F X , Gößner I , Tascher G , et al . A cytosolic surveillance mechanism activates the mitochondrial UPR [J]. Nature , 2023 , 618 ( 7966 ): 849 - 854 .
Uoselis L , Lindblom R , Lam W K , et al . Temporal landscape of mitochondrial proteostasis governed by the UPR(mt) [J]. Sci Adv , 2023 , 9 ( 38 ): eadh8228 .
Lu H D , Wang X L , Li M , et al . Mitochondrial unfolded protein response and integrated stress response as promising therapeutic targets for mitochondrial diseases [J]. Cells , 2023 , 12 ( 1 ): 20 .
Liu B Y , Wu J N , Yan S Y , et al . Electroacupuncture vs prucalopride for severe chronic constipation: a multicenter, randomized, controlled, noninferiority trial [J]. Am J Gastroenterol , 2021 , 116 ( 5 ): 1024 - 1035 .
Zheng H , Liu Z S , Zhang W , et al . Acupuncture for patients with chronic functional constipation: a randomized controlled trial [J]. Neurogastroenterol Motil , 2018 , 30 ( 7 ): e13307 .
张微 . 针刺俞募穴改善FC小鼠胃肠传输功能的GDNF-PI 3 K-AKT信号转导机制研究 [D]. 成都 : 成都中医药大学 , 2016 .
Zhang W . Research on GDNF-PI3K-AKT ignal pathway in FC mice with acupuncture back-Shu and front-mu points improving gastrointestinal transmitting function (in Chinese) [D]. Chengdu : Chengdu University of TCM , 2016 .
Yao J P , Yan X Y , Li Y Q , et al . Altered gut microbial profile is associated with differentially expressed fecal microRNAs in patients with functional constipation [J]. Front Microbiol , 2024 , 14 : 1323877 .
Wang L , Xi M H , Cao W , et al . Electroacupuncture alleviates functional constipation by upregulating host-derived miR-205-5p to modulate gut microbiota and tryptophan metabolism [J]. Front Microbiol , 2025 , 16 : 1517018 .
Zhang Q , Zhong D , Sun R , et al . Prevention of loperamide induced constipation in mice by KGM and the mechanisms of different gastrointestinal tract microbiota regulation [J]. Carbohydr Polym , 2021 , 256 : 117418 .
Yan X Y , Ma P T , Wang W , et al . Piezo knockdown reduces 5-hydroxytryptamine release from enterochromaffin cells and exacerbates intestinal dyskinesia in mice with functional constipation [J]. Int J Mol Med , 2025 , 56 ( 5 ): 1 - 17 .
余曙光 , 徐斌 . 实验针灸学 [M]. 北京 : 人民卫生出版社 , 2012 : 269 - 277 .
Yu S G , Xu B . Experimental acupuncturology (in Chinese) [M]. Beijing : People’s Medical Publishing House , 2012 : 269 - 277 .
Lewis S J , Heaton K W . Stool form scale as a useful guide to intestinal transit time [J]. Scand J Gastroenterol , 1997 , 32 ( 9 ): 920 - 924 .
王路 , 席梦含 , 张微 , 等 . 基于机械敏感离子通道探讨电针改善功能性便秘小鼠胃肠动力的机制 [J]. 中华中医药杂志 , 2023 , 38 ( 11 ): 5237 - 5243 .
Wang L , Xi M H , Zhang W , et al . Mechanism research of electroacupuncture improving gastrointestinal motility in functional constipation mice based on mechanosensitive ion channels (in Chinese) [J]. China Journal of Traditional Chinese Medicine and Pharmacy , 2023 , 38 ( 11 ): 5237 - 5243 .
席梦含 , 王路 , 张微 , 等 . 电针对功能性便秘小鼠肠道神经元自噬水平的影响 [J]. 中国针灸 , 2023 , 43 ( 11 ): 1279 - 1286 .
Xi M H , Wang L , Zhang W , et al . Effect of electroacupuncture on enteric neuronal autophagy in functional constipation mice (in Chinese) [J]. Chinese Acupuncture & Moxibustion , 2023 , 43 ( 11 ): 1279 - 1286 .
Bhardwaj J , Wan D Q , Koenig M K , et al . Impaired gastric emptying and small bowel transit in children with mitochondrial disorders [J]. J Pediatr Gastroenterol Nutr , 2012 , 55 ( 2 ): 194 - 199 .
Finsterer J , Frank M . Gastrointestinal manifestations of mitochondrial disorders: a systematic review [J]. Ther Adv Gastroenterol , 2017 , 10 ( 1 ): 142 - 154 .
Martin-Cano F E , Camello-Almaraz C , Acuña-Castroviejo D , et al . Age-related changes in mitochondrial function of mouse colonic smooth muscle: beneficial effects of melatonin [J]. J Pineal Res , 2014 , 56 ( 2 ): 163 - 174 .
Camilleri M , Carlson P , Zinsmeister A R , et al . Mitochondrial DNA and gastrointestinal motor and sensory functions in health and functional gastrointestinal disorders [J]. Am J Physiol Gastrointest Liver Physiol , 2009 , 296 ( 3 ): G510 - G516 .
Inigo J R , Chandra D . The mitochondrial unfolded protein response (UPRmt): shielding against toxicity to mitochondria in cancer [J]. J Hematol Oncol , 2022 , 15 ( 1 ): 98 .
Katiyar A , Fujimoto M , Tan K , et al . HSF1 is required for induction of mitochondrial chaperones during the mitochondrial unfolded protein response [J]. FEBS Open Bio , 2020 , 10 ( 6 ): 1135 - 1148 .
梁星琛 , 白璐璐 , 周永学 , 等 . 基于 “脑-肠轴” 线粒体未折叠蛋白反应研究济川煎对慢传输型便秘的作用机制 [J]. 中国中西医结合杂志 , 2022 , 42 ( 1 ): 75 - 82 .
Liang X C , Bai L L , Zhou Y X , et al . Mechanism of jichuanjian for treatment of slow transit constipation based on brain-gut axis mitochondrial unfolded protein reaction (in Chinese) [J]. Chinese Journal of Integrated Traditional and Western Medicine , 2022 , 42 ( 1 ): 75 - 82 .
Berger E , Rath E , Yuan D T , et al . Mitochondrial function controls intestinal epithelial stemness and proliferation [J]. Nat Commun , 2016 , 7 : 13171 .
王飞 , 李亚楠 , 张小蕾 , 等 . 电针调节PI3K/AKT/mTOR信号通路激活帕金森大鼠脑组织自噬改善行为学反应机制 [J]. 中国临床解剖学杂志 , 2023 , 41 ( 6 ): 709 - 715 .
Wang F , Li Y N , Zhang X L , et al . Electroacupuncture activates autophagy and improves behavioral responses mechanism in Parkinson’s rats brain by regulating PI3K/AKT/mTOR signaling pathway (in Chinese) [J]. Chinese Journal of Clinical Anatomy , 2023 , 41 ( 6 ): 709 - 715 .
余致韵 , 杨文淼 , 许明敏 , 等 . 电针对抑郁模型小鼠海马神经元线粒体自噬的影响 [J]. 北京中医药大学学报 , 2025 , 48 ( 9 ): 1310 - 1323 .
Yu Z Y , Yang W M , Xu M M , et al . Effects of electroacupuncture on mitophagy in hippocampal neurons of mouse depression model (in Chinese) [J]. Journal of Beijing University of Traditional Chinese Medicine , 2025 , 48 ( 9 ): 1310 - 1323 .
吴阳阳 , 刘明江 , 殷韶杰 , 等 . 针刺对溃疡性结肠炎大鼠氧化应激和内质网应激的影响 [J]. 针刺研究 , 2020 , 45 ( 1 ): 8 - 14 .
Wu Y Y , Liu M J , Yin S J , et al . Acupuncture reduce colonic inflammation by suppressing oxidative stress and endoplasmic reticulum stress in rats with ulcerative colitis (in Chinese) [J]. Acupuncture Research , 2020 , 45 ( 1 ): 8 - 14 .
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