甘草泻心汤联合美沙拉秦缓释颗粒对急性期溃疡性结肠炎大鼠肠黏膜屏障的作用研究

Experimental research of Gancao Xiexin decoction combined with Mesalachin slow-release granules on intestinal mucosal barrier of ulcerative colitis rats in acute phase

摘要:

目的 探讨甘草泻心汤联合美沙拉秦缓释颗粒治疗急性期溃疡性结肠炎的可能作用机制。方法 将50只健康的雄性SD大鼠随机分为5组［空白组、模型组、甘草泻心汤组、美沙拉秦组及甘草泻心汤联合美沙拉秦组（以下简称联合组）］，每组10只。除空白组外，其余大鼠均采用3%硫酸葡聚糖钠盐（分子量40 000）水溶液自由饮用7 d复制急性期溃疡性结肠炎大鼠模型。模型复制成功后，空白组和模型组采用纯净水干预，其余3组分别采用甘草泻心汤、美沙拉秦混悬液、甘草泻心汤联合美沙拉秦混悬液灌胃治疗14 d。干预治疗后观察大鼠的一般情况，测量各组大鼠体重并进行疾病活动指数（DAI）评分，采用HE染色法观察结肠组织病理改变并进行组织学损伤指数（HI）评分，酶联免疫吸附试验（ELISA）检测血清二胺氧化酶（DAO）和D-乳酸（D-LA）的含量，显色基质鲎试剂定量检测大鼠血清细菌内毒素，Western blotting检测ZO-1和Occludin蛋白表达。结果 与空白组比较，模型组大鼠DAO和D-LA含量及细菌内毒素定量均升高（P <0.05），ZO-1和Occludin蛋白表达降低（P <0.05）。与模型组比较，甘草泻心汤组、美沙拉秦组及联合组的大鼠DAI评分和HI评分降低（P <0.05），DAO和D-LA含量及细菌内毒素定量均降低（P <0.05），ZO-1和Occludin蛋白表达升高（P <0.05）。各治疗组间比较，联合组干预治疗后疗效优于甘草泻心汤组和美沙拉秦组。结论 甘草泻心汤联合美沙拉秦缓释颗粒对急性期溃疡性结肠炎大鼠的肠黏膜屏障具有一定的修复功能，作用机制可能与ZO-1和Occludin蛋白表达有关，进而影响肠黏膜的通透性与完整性。

关键词:溃疡性结肠炎;甘草泻心汤;肠黏膜屏障;紧密连接蛋白

Abstract:

Objective To explore the possible mechanism of action of Gancao Xiexin decoction(GCXX decoction) combined with Mesalachin extended-release granules in the treatment of acute phase ulcerative colitis.Methods Fifty healthy male SD rats were randomly divided into five groups (blank group, model group, GCXX decoction group, Mesalachin group, GCXX decoction combined with Mesalachin group here after referred to as the combined group). Except for 10 rats in the blank group, all other rats were used 3% dextran sulfate nano-salt (molecular weight 40 000) aqueous solution for 7 d to establish the acute phase UC rat model. The successfully modeled rats were randomly divided into the model group, GCXX decoction group, Mesalachin group and combined group, with 10 rats in each group. The blank group and the model group were treated with pure water, and the remaining three groups were treated with GCXX decoction, Mesalachin suspension, and GCXX decoction combined with Mesalachin suspension by gavage for 14 d. After the intervention, the general condition of the rats was observed, the body mass of each group was measured and the disease activity index (DAI) was scored, and the histological changes of the colon were observed by HE staining and the histological damage index (HI) was scored, the levels of serum diamine oxidase (Diamine Oxidase, DAO）and D-lactic acid (D-Lactic, D-LA) were measured by ELISA, and the expression of tight junction proteins (ZO-1) and occludin (Occludin) were detected by Western blotting analysis.Results Compared with the blank group, rats in the model group had increased DAO and D-LA content (P < 0.05), increased bacterial endotoxin quantification (P < 0.05), and decreased ZO-1 and Occludin protein expression (P < 0.05). Compared with the model group, the rats in the GCXX decoction group, Mesalachin group, and combined groups had lower DAI scores (P < 0.05), lower HI scores (P < 0.05), lower DAO and D-LA contents (P < 0.05) and lower bacterial endotoxin quantification (P < 0.05), and higher ZO-1 and Occludin protein expressions (P < 0.05). In comparison between treatment groups, the efficacy of the combined group was better than that of the GCXX decoction group and Mesalachin group after the intervention.Conclusion GCXX decoction combined with Mesalachin slow-release granules has a certain repair function on the intestinal mucosal barrier of UC rats in the acute phase, and the mechanism of action may be related to ZO-1 and Occludin protein expression, thus affecting the permeability and integrity of the intestinal mucosa.

Key words:colitis, ulcerative;Gancao Xiexin decoction;intestinal mucosal barrier;tight junctions