电针调节巨噬细胞极化改善大鼠溃疡性结肠炎机制的研究。

Hui-Chao Xu, Rong-Lin Wu, Zi-Wen Jiang, Hai-Jun Wang, Yu-Xia Cao, Jian-Heng Hao, Rang-Qian Li, Lai-Xi Ji
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引用次数: 0

摘要

目的:观察电针长冰汤对大鼠溃疡性结肠炎(UC)的缓解作用及对结肠巨噬细胞极化的调节作用,探讨其改善UC的作用机制,西药组(6只)。用5%右旋糖酐硫酸钠(DSS)溶液饮水7天,治疗期间饮用1%DSS溶液,建立UC大鼠模型。模型建立7天后,电针治疗(10Hz/50Hz,20min)于“中丸”(CV12)、双侧“天舒”(ST25)和“上巨虚”(ST37)3d,西药组大鼠灌胃给予美沙拉秦混悬液(200mg/kg)3d。评估疾病活动指数(DAI)评分。HE染色后检测结肠的形态学变化和炎症细胞浸润,并评估病理评分。采用ELISA法检测血清中肿瘤坏死因子α(TNF-α)、白细胞介素-1β、白介素-2和白细胞介素-10的含量。用荧光双染法和蛋白质印迹法分别检测M1和M2巨噬细胞标志物一氧化氮合酶(iNOS)和精氨酸酶1(Arg1)的蛋白表达。实时定量PCR检测iNOS和Arg1mRNA的表达。结果:与正常组相比,模型组大鼠结肠组织病理损伤程度和炎症细胞浸润增加,体重增加减慢,结肠长度、脾脏重量、血清抗炎因子IL-2和IL-10含量下降,结肠Arg1/CD68荧光阳性表达,结论:长冰汤电针可通过调节结肠巨噬细胞极化、抑制M1巨噬细胞生成、促进M2巨噬细胞生成等途径改善大鼠UC。
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Study on the mechanism of electroacupuncture regulating macrophage polarization to improve ulcerative colitis in rats.

Objectives: To investigate the effect of electroacupuncture(EA) at "Changbing Decoction" on alleviating ulcerative colitis (UC) and regulating the polarization of colonic macrophages in rats, so as to explore its mechanisms underlying improvement of UC.

Methods: Twenty-six male SD rats were randomly divided into 4 groups:normal group(6 rats), model group(8 rats), EA group(6 rats), and western medication group(6 rats). The rat model of UC was established by using 5% dextran sulfate sodium (DSS) solution drinking water for 7 days, followed by drinking 1% DSS solution during treatment period. After 7-day model establishment, EA treatment(10 Hz/50 Hz, 20 min) was applied to "Zhongwan"(CV12), bilateral "Tianshu"(ST25) and "Shangjuxu"(ST37) for 3 d, and rats in the western medication group were given mesalazine suspension(200 mg/kg) by gavage for 3 d. The body weight, spleen weight and colon length of rats were measured. The disease activity index (DAI) score was evaluated. The morphological changes and inflammatory cell infiltration of colon were detected after HE staining and pathological scores were eva-luated. The contents of tumor necrosis factor α(TNF-α), interleukin(IL)-1β, IL-2 and IL-10 in serum were detected by ELISA. The protein expressions of M1 and M2 macrophage markers nitric oxide synthase (iNOS) and arginase 1(Arg1) were detected by fluorescence double staining and Western blot, respectively. Quantitative real-time PCR was used to detect iNOS and Arg1 mRNA expressions.

Results: Compared with the normal group, rats in the model group had increased pathological damage degree and inflammatory cell infiltration in the colon tissue, slowed-down body weight gain, decreased colon length, spleen weight, serum anti-inflammatory factors IL-2 and IL-10 contents, colonic Arg1/CD68 fluorescence positive expression, and Arg1 protein and mRNA expressions(P<0.01, P<0.05), as well as increased DAI scores, colon histopathological scores, contents of serum pro-inflammatory factors TNF-α and IL-1β, colonic iNOS/CD68 fluorescence positive expression, iNOS protein and mRNA expressions(P<0.01). Compared with the model group, the above indicators were significantly improved in rats of the EA group and the western medication group(P<0.01, P<0.05).

Conclusions: EA of "Changbing Decoction" can improve UC of rats by regulating the polarization of colonic macrophages, inhibiting the generation of M1 macrophages and promoting the generation of M2 macrophages.

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