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电针对调节巨噬细胞极化改善溃疡性结肠炎大鼠作用机制的研究。

Study on the mechanism of electroacupuncture regulating macrophage polarization to improve ulcerative colitis in rats.

机构信息

College of Acupuncture-moxibustion and Tuina, Chengdu University of Traditional Chinese Medicine, Chengdu 610075, China.

The Second Clinical College, Shanxi University of Chinese Medicine, Jinzhong 030619, Shanxi Province.

出版信息

Zhen Ci Yan Jiu. 2023 Oct 25;48(10):1033-1040. doi: 10.13702/j.1000-0607.20230286.

Abstract

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(<0.01, <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(<0.01). Compared with the model group, the above indicators were significantly improved in rats of the EA group and the western medication group(<0.01, <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.

摘要

目的

观察电针“肠痈方”对溃疡性结肠炎(UC)大鼠的作用及其对结肠巨噬细胞极化的影响,探讨其改善 UC 的作用机制。

方法

26 只雄性 SD 大鼠随机分为 4 组:正常组(6 只)、模型组(8 只)、电针组(6 只)和西药组(6 只)。采用 5%葡聚糖硫酸钠(DSS)溶液饮水 7 天建立 UC 大鼠模型,造模成功后继续给予 1% DSS 溶液饮水 7 天进行造模,造模结束后电针治疗 3 天,电针“中脘”(CV12)、双侧“天枢”(ST25)和“上巨虚”(ST37),10 Hz/50 Hz,20 min/次;西药组给予美沙拉嗪混悬液(200 mg/kg)灌胃 3 天。观察大鼠体质量、脾脏质量和结肠长度,评估疾病活动指数(DAI)评分,HE 染色观察结肠组织形态学改变和炎症细胞浸润情况,评估病理损伤评分,ELISA 法检测血清肿瘤坏死因子-α(TNF-α)、白细胞介素(IL)-1β、IL-2 和 IL-10 含量,荧光双染法检测 M1 和 M2 型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,实时荧光定量 PCR 法检测 iNOS 和 Arg1mRNA 表达。

结果

与正常组比较,模型组大鼠结肠组织病理损伤程度和炎症细胞浸润加重,体质量增长缓慢,结肠长度、脾脏质量、血清抗炎因子 IL-2 和 IL-10 含量、结肠 Arg1/CD68 荧光阳性表达、Arg1 蛋白和 mRNA 表达降低(<0.01,<0.05),DAI 评分、结肠组织病理损伤评分、血清促炎因子 TNF-α和 IL-1β含量、结肠 iNOS/CD68 荧光阳性表达、iNOS 蛋白和 mRNA 表达升高(<0.01);与模型组比较,电针组和西药组上述指标均有不同程度改善(<0.01,<0.05)。

结论

电针“肠痈方”可能通过调节结肠巨噬细胞极化,抑制 M1 型巨噬细胞生成、促进 M2 型巨噬细胞生成而改善 UC 大鼠的病情。

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