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1.广州中医药大学第二临床医学院,广州 510006
2.中医证候全国重点实验室,广州中医药大学第二附属医院,广州 510000
3.广东省中医证候临床研究重点实验室,广州中医药大学第二附属医院,广州 510000
刘华桢,E-mail:hz.liu@gzucm.edu.cn
收稿:2026-02-25,
修回:2026-06-10,
网络首发:2026-07-31,
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李学佳,李泳丹,刘昭霖,等.基于TRAF2/NF-κB通路探讨电针调控CD8⁺记忆T细胞抑制银屑病复发的作用机制[J].针刺研究,XXXX,XX(XX):1-13.
Li Xuejia,Li Yongdan,Liu Zhaolin,et al.Mechanism of electroacupuncture inhibiting psoriasis recurrence by regulating CD8+ memory T cells based on the TRAF2/NF-κB signaling pathway[J].Acupuncture Research,DOI:10.13702/j.1000⁃0607.20260218. DOI: 10.13702/j.1000-0607.20260218.
李学佳,李泳丹,刘昭霖,等.基于TRAF2/NF-κB通路探讨电针调控CD8⁺记忆T细胞抑制银屑病复发的作用机制[J].针刺研究,XXXX,XX(XX):1-13. DOI: 10.13702/j.1000-0607.20260218.
Li Xuejia,Li Yongdan,Liu Zhaolin,et al.Mechanism of electroacupuncture inhibiting psoriasis recurrence by regulating CD8+ memory T cells based on the TRAF2/NF-κB signaling pathway[J].Acupuncture Research,DOI:10.13702/j.1000⁃0607.20260218. DOI: 10.13702/j.1000-0607.20260218. DOI:
目的
2
探讨电针对银屑病复发模型小鼠的治疗作用,并从记忆T细胞分化及肿瘤坏死因子受体相关因子2(TRAF2)/核因子κB(NF-κB)信号通路角度探讨其作用机制。
方法
2
将BALB/c小鼠分批用于复发模型及机制验证实验。复发模型设空白对照组、模型组、电针组、甲氨蝶呤组及电针+甲氨蝶呤组;验证实验增设电针+NF-κB抑制剂组和电针+NF-κB激动剂组。采用背部涂抹咪喹莫特(IMQ)诱导银屑病复发模型。电针干预选取“曲池”“足三里”,疏密波,频率2 Hz/30 Hz,强度1 mA,每日1次,连续7 d,每次15 min。观察皮损变化并进行银屑病皮损面积及严重程度指数(PASI)评分;苏木精-伊红(HE)染色观察皮损处皮肤组织病理学改变;ELISA法检测皮损处皮肤组织中白细胞介素(IL)-6、IL-17A、IL-15及肿瘤坏死因子-α(TNF-α)含量;实时荧光定量PCR法检测皮损处皮肤组织IL-15、C-X-C模体趋化因子(CXCL)9、CXCL10、C-X-C趋化因子受体3(CXCR3)mRNA表达;流式细胞术检测皮损组织中CD8⁺组织常驻记忆T细胞(T
RM
)及CD4
+
T
RM
比例;Western blot法检测皮损处皮肤组织中磷酸化NF-κB p65(p-NF-κB p65)、NF-κB p52、磷酸化TRAF2(p-TRAF2)、磷酸化p38丝裂原活化蛋白激酶(p-p38 MAPK)、蛋白表达。
结果
2
与空白对照组相比,模型组PASI,IL-6、TNF-α、IL-17A含量、CD8⁺及CD4⁺ T
RM
比例,IL-15、CXCL9、CXCL10、CXCR3 mRNA表达,p-TRAF2、p-NF-κB p65、NF-κB p52表达均显著升高(
P
<
0.01,
P
<
0.05)。与模型组相比,电针及电针+甲氨蝶呤组上述指标均显著逆转(
P
<
0.01,
P
<
0.05)。与甲氨蝶呤组相比,两电针组上述指标(除IL-17A,CD4⁺T
RM
比例外)均显著降低(
P
<
0.01,
P
<
0.05)。与电针组相比,电针+甲氨蝶呤组CD8⁺CD103⁺T
RM
、CXCL9、CXCL10 mRNA及p-NF-κB p65表达进一步降低(
P
<
0.01,
P
<
0.05)。验证实验中,与模型组相比,电针及电针+NF-κB抑制剂组皮损、炎性因子、CD8⁺CD103⁺ T
RM
、CD4
+
T
RM
比例,p-TRAF2、p-NF-κB p65表达,CXCL9、CXCL10、CXCR3 mRNA表达均显著改善(
P
<
0.01,
P
<
0.05);与电针组相比,电针+抑制剂组上述指标(除PASI)进一步降低(
P
<
0.05,
P
<
0.01),电针+激动剂组IL-6、TNF-α、IL-17A、IL-15含量,CD8⁺CD103⁺T
RM
比例,p-TRAF2、p-NF-κB p65蛋白表达显著升高(
P
<
0.01,
P
<
0.05)。
结论
2
电针能有效改善银屑病复发症状,其作用机制可能与抑制TRAF2/NF-κB信号通路活化,特异性下调CD8⁺记忆T细胞水平有关。
Objective
2
To investigate the therapeutic effect of electroacupuncture(EA) on mice with recurrent psoriasis, so as to explore its mechanism from the perspective of memory T cell differentiation and the tumor necrosis factor receptor-associated factor 2 (TRAF2)/nuclear factor-κB (NF-κB) signaling pathway.
Methods
2
BALB/c mice were divided into batches for establishment of recurrent psoriasis model and mechanism verification experiments. The recurrence experiment included blank control group, model group, EA group, methotrexate(MTX) group, and EA + MTX group. The verification experiment additionally set up EA + NF-κB inhibitor group and EA + NF-κB agonist group. The recurrent psoriasis model was induced by topical imiquimod (IMQ) application on the back skin. EA was applied at “Quchi” (LI11) and “Zusanli” (ST36) with sparse-dense wave, frequency of 2 Hz/30 Hz and intensity of 1 mA, once daily for 7 consecutive days, 15 min each time. Lesion changes were observed and psoriasis area and severity index (PASI) scores were evaluated. HE staining was adopted to observe histopathological alterations of lesional skin tissues. ELISA was used to detect the contents of interleukin (IL)-6, IL-17A, IL-15 and tumor necrosis factor-α (TNF-α) in lesional skin tissues. Real-time quantitative PCR was performed to detect mRNA expressions of IL-15, C-X-C motif chemokine ligand (CXCL) 9, CXCL10 and C-X-C chemokine receptor 3 (CXCR3) in lesional skin tissue. Flow cytometry was applied to detect the proportions of CD8
+
tissue-resident memory T cells (T
RM
) and CD4
+
T
RM
in skin lesions. Western blot was used to test protein expression of phosphorylated NF-κB p65 (p-NF-κB p65), NF-κB p52, phosphorylated TRAF2 (p-TRAF2), and phosphorylated p38 mitogen-activated protein kinase (p-p38 MAPK) in lesional skin tissues.
Results
2
Compared with the blank control group, the model group exhibited significantly increased PASI score, contents of IL-6, TNF-α and IL-17A, proportio
ns of CD8
+
T
RM
and CD4
+
T
RM
, mRNA expression of IL-15, CXCL9, CXCL10, CXCR3, as well as protein expressions of p-TRAF2, p-NF-κB p65 and NF-κB p52 (
P
<
0.01,
P
<
0.05). Compared with the model group, all above indicators were markedly reversed in the EA group and EA + MTX group (
P
<
0.01,
P
<
0.05). Except for the proportion of CD4
+
T
RM
and IL-17A, the remaining indicators were significantly lower in the two EA-related groups than those in the MTX group (
P
<
0.01,
P
<
0.05). Compared with the EA group, the EA + MTX group showed further decreased proportions of CD8
+
CD103
+
T
RM
, down-regulated mRNA expressions of CXCL9, CXCL10 and reduced p-NF-κB p65 protein expression (
P
<
0.01,
P
<
0.05). In verification experiments, compared with the model group, the EA group and EA + NF-κB inhibitor group presented obvious improvements in skin lesions, inflammatory factors, proportions of CD8
+
CD103
+
T
RM
and CD4
+
T
RM
, protein expressions of p-TRAF2, p-NF-κB p65, and mRNA levels of CXCL9, CXCL10, CXCR3 (
P
<
0.05,
P
<
0.01). Compared with the EA group, these indicators were further decreased in the EA + inhibitor group (
P
<
0.01,
P
<
0.05); in the EA + agonist group, the contents of IL-6, TNF-α, IL-17A, IL-15, proportion of CD8
+
CD103
+
T
RM
, and protein expressions of p-TRAF2, p-NF-κB p65 were significantly elevated (
P
<
0.01,
P
<
0.05).
Conclusion
2
EA can effectively alleviate symptoms of recurrent psoriasis. Its underlying mechanism may be related to inhibiting the activation of the TRAF2/NF-κB signaling pathway and specifically down-regulating the level of CD8
+
memory T cells.
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