左金丸对结肠炎小鼠外周血CD45RA^+CD62L^+CCR7^+T细胞及其CD4^+、CD8^+亚群的影响

Effects of Zuojin Pill on CD45RA^+CD62L^+CCR7^+T cells and CD4^+,CD8^+subsets of peripheral blood in colitis mice

目的:探讨左金丸对葡聚糖硫酸钠(DSS)诱导小鼠结肠炎外周血中CD45RA+CD62L+CCR7+T细胞及其亚群的影响。方法:将40只雄性BALB/c小鼠随机平均分为正常组、模型组、左金丸组、美沙拉嗪组。除正常组外,其余各组均用3%DSS自由饮水7d诱导小鼠结肠炎,第8天开始予左金丸、美沙拉唑肠溶片治疗7d。观察小鼠一般情况及结肠病理损伤变化,采用流式细胞术测定外周血中CD45RA+CD62L+CCR7+T细胞及其CD4+、CD8+亚群水平,Western blotting检测结肠组织中磷脂酰肌醇3-激酶(PI3K)、蛋白激酶B(Akt)和磷酸化蛋白激酶B(p-Akt)蛋白表达水平并计算p-Akt/Akt值。结果:与模型组比较,左金丸组可有效改善DSS诱导结肠炎小鼠体质量、结肠长度、肠重指数和黏膜镜下评分(P<0.05,P<0.01),显著升高外周血中CD45RA+CD62L+CCR7+T细胞及其CD4+、CD8+亚群水平(P<0.01,P<0.05),同时伴见结肠组织中PI3K、p-Akt蛋白表达量显著降低(P<0.01,P<0.05)和p-Akt/Akt比值显著下降(P<0.01)。结论:左金丸可通过抑制PI3K/Akt信号活化调节CD45RA+CD62L+CCR7+T细胞及其CD4+、CD8+亚群水平而达到治疗DSS诱导小鼠结肠炎的目的。

Objective: Investigate the effect of Zuojin Pill(ZJP) on CD45 RA+CD62 L+CCR7+T cells and their subsets in peripheral blood of mice with colitis induced by dextran sulfate sodium(DSS). Methods: A total of 40 male BALB/c mice were randomly divided into normal group, model group, ZJP group and mesalazine group. Except for the normal group, the other groups were treated with 3%DSS free drinking water for 7 days to induce colitis in mice, and ZJP and Mesalazine Entercoated Tablets were given for 7 days on the 8 th day. The general situation of mice and the pathological changes of colon were observed. The levels of CD45 RA+CD62 L+CCR7+T cells and their CD4+ and CD8+ subsets in peripheral blood were measured by flow cytometry. The expression levels of phosphatidylinositol 3-kinase(PI3 K), protein kinase B(Akt) and phosphorylated protein kinase B(p-Akt) in colon tissue were detected by Western blotting and the value of p-Akt/Akt was calculated. Results: Compared with the model group, ZJP group could effectively improve the body weight, colon length, intestinal weight index and mucosal endoscopic score of DSS-induced colitis mice(P<0.05, P<0.01). The level of CD45 RA+CD62 L+CCR7+T cells and their CD4+ and CD8+ subsets in peripheral blood were significantly increased(P<0.01, P<0.05), and the expression of PI3 K and p-Akt protein in colon tissues were significantly decreased(P<0.01, P<0.05). The ratio of p-Akt/Akt was significantly decreased(P<0.01). Conclusion: ZJP can treat colitis induced by DSS in mice by inhibiting the activation of PI3 K/Akt signal and regulating the level of CD45 RA+CD62 L+CCR7+T cells and their CD4+ and CD8+ subsets.