[电针对急性溃疡性结肠炎大鼠结肠自噬和 AMPK/mTOR 信号通路的影响]

Rong-Lin Wu, Hui-Chao Xu, Zi-Wen Jiang, Hai-Jun Wang, Yu-Xia Cao, Fan-Min Kong, Ai-Ai Dong, Rang-Qian Li, Lai-Xi Ji
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引用次数: 0

摘要

目的观察电针(EA)在 "中万"(CV12)、"天枢"(ST25在 "中丸"(CV12)、"天枢"(ST25)和 "尚书"(ST37)(治疗肠道疾病的穴位处方 "长炳方")。方法:研究32只雄性SD大鼠,观察 "天枢"(ST25)和 "上巨虚"(ST37)对溃疡性结肠炎(UC)大鼠自噬和AMPK/mTOR信号通路表达的影响,探讨其改善UC的机制:将32只雄性SD大鼠随机分为对照组、模型组、药物组和EA组,每组8只。通过自由饮用5%葡聚糖硫酸钠盐溶液建立UC模型,为期7天。EA刺激(10赫兹/50赫兹)连续3天,每天一次,每次20分钟。药物组大鼠灌胃美沙拉嗪混悬液(200 毫克/千克),每天一次,共 3 次。每天一次,共 3 次。记录大鼠的一般情况以计算疾病活动指数(DAI)0-4分)。通过 HE 染色观察结肠的组织形态学变化。血清白细胞介素6(IL-6)、肿瘤坏死因子-α(TNF-α)和IL-10的水平由ELIS法测定。和 IL-10 的水平。荧光定量 PCR 检测 LC3B 和 p62 的 mRNA 表达。Western blot检测结肠组织中LC3B、p62和AMPK/mTOR通路相关蛋白的表达水平:结果:与对照组相比,DAI评分、血清中IL-6和TNF-α的含量、p62蛋白和mRNA的表达水平、p-mTOR/mTOR的比值均显著升高(PPPα和IL-10的含量)、药物组和 EA 组 p62 蛋白和 mRNA、LC3B mRNA、p-mTOR/mTOR、LC3BⅡ/LC3BⅠ、p-AMPK/AMPK 的表达均发生逆转(PPPPPC结论穴位敷贴能改善UC大鼠的症状,这可能与其促进结肠自噬、提高AMPK磷酸化水平和降低mTOR磷酸化水平有关。
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[Effect of electroacupuncture on colonic autophagy and AMPK/mTOR signaling pathway in rats with acute ulcerative colitis].

Objective: To observe the effect of electroacupuncture (EA) at "Zhongwan" (CV12), "Tianshu" (ST25) and "Shangjuxu" (ST37) (an acupoint prescription "Changbingfang" for treatment of intestinal disorders) on autophagy and expression of AMPK/mTOR signaling pathway in rats with ulcerative colitis (UC), so as to explore its mechanism underlying improvement of UC.

Methods: Thirty-two male SD rats were randomly divided into control, model, medication and EA groups, with 8 rats in each group. The UC model was established by free drinking of 5% dextran sulfate sodium salt solution for 7 days. EA stimulation (10 Hz/50 Hz) was delivered to CV12, ST25 and ST37 for 20 min, once a day for 3 consecutive days. Rats of the medication group received gavage of mesalazine suspension (200 mg/kg) once a day, 3 times in total. The rats' general conditions were recorded for calculating the disease activity index (DAI) score (0-4 points). Histomorphological changes of colon were observed via HE staining. The levels of serum interleukin 6 (IL-6), tumor necrosis factor-α (TNF-α) and IL-10 were measured by ELISA. The mRNA expressions of LC3B and p62 were tested by fluorescence quantitative PCR. Western blot was used to detect the expression levels of LC3B, p62 and AMPK/mTOR pathway related proteins in colon tissues.

Results: Compared with the control group, the DAI score, contents of serum IL-6 and TNF-α, the expression levels of p62 protein and mRNA, ratio of p-mTOR/mTOR were significantly increased (P<0.01); while the content of serum IL-10, the expression levels of LC3B mRNA, ratio of LC3BⅡ/LC3BⅠ and p-AMPK/AMPK were decreased (P<0.01, P<0.05) in the model group. Relevant to the model group, modeling-induced increases of DAI score, serum IL-6, TNF-α and IL-10 contents, expressions of p62 protein and mRNA, LC3B mRNA, ratio of p-mTOR/mTOR, LC3BⅡ/LC3BⅠ and p-AMPK/AMPK were reversed in both medication and EA groups (P<0.01, P<0.05). The effect of EA was apparently superior to that of mesalazine in up-regulating ratio of LC3BⅡ/LC3BⅠ and p-AMPK/AMPK, p62 mRNA expression (P<0.01, P<0.05), and in down-regulating ratio of p-mTOR/mTOR (P<0.05). H.E. staining showed severe damage of the colonic mucosal barrier with infiltration of a large number of inflammatory cells in the model group, which was milder in medication and EA groups.

Conclusion: EA of acupoint recipe "Changbingfang" can improve the symptoms in UC rats, which may be related to its functions in promoting colonic autophagy, increasing AMPK phosphorylation level, and decreasing mTOR phosphorylation level.

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