【摘要】 目的:探讨茵陈苓桂术甘汤对非酒精性脂肪性肝病(nonalcoholic fatty liver disease, NAFLD)小鼠Th17/Treg细胞免疫平衡的调控作用，为其临床应用提供依据。方法:将SPF级雄性C57BL/6小鼠随机分为空白组、模型组、茵陈苓桂术甘汤低剂量组、茵陈苓桂术甘汤中剂量组和茵陈苓桂术甘汤高剂量组。采用高脂饲料诱导NAFLD小鼠模型，造模成功后药物组分别予相应浓度药物灌胃，空白组和模型组予等量的蒸馏水灌胃，1次/d,共4周，观察不同方式治疗后小鼠体重及肝组织病理学变化，通过流式细胞术检测小鼠肠系膜淋巴结辅助性T细胞17(Th17细胞)、调节性T细胞(Treg细胞)水平，以及Th17/Treg细胞比率；并通过RT-qPCR检测二者相关细胞因子IL-17、IL-10的mRNA表达。结果:与空白组比较，模型组小鼠体重明显增加(P<0.01),肝细胞出现空泡样变性及脂肪变性，炎性细胞浸润明显；经治疗4周后，茵陈苓桂术甘汤各剂量组小鼠体重明显降低(P<0.05,P<0.01);肝组织空泡样变性、脂肪样变性及炎性细胞浸润均明显减轻。机制方面，与空白组比较，模型组小鼠肠系膜淋巴结Th17占比显著升高(P<0.01),其相关细胞因子IL-17的mRNA表达水平显著升高(P<0.05);而Treg细胞占比显著降低(P<0.05),其相关细胞因子IL-10的mRNA表达水平降低(P<0.01),Th17/Treg细胞比率显著升高，差异有统计学意义(P<0.01)。茵陈苓桂术甘汤组与模型组比较，小鼠肠系膜淋巴结Th17占比显著降低(P<0.01),其相关细胞因子IL-17的mRNA水平显著降低(P<0.01);而Treg细胞占比显著升高(P<0.01),其相关细胞因子IL-10的mRNA表达水平显著升高(P<0.01);Th17/Treg显著降低，差异有统计学意义(P<0.01)。结论:茵陈苓桂术甘汤可以显著改善NAFLD小鼠肝细胞的脂肪变性及炎性细胞浸润，其作用机制与调节Th17/Treg细胞免疫平衡有关。更多还原

【Abstract】 Objective:To investigate the modulating effect of Yinchen Linggui Zhugan Decoction(YLZD) on Th17/Treg immune balance in nonalcoholic fatty liver disease(NAFLD) mice, providing a scientific basis for its treatment of NAFLD. Methods: SPF-grade male C57BL/6 mice were randomly divided into a control group, model group, low-dose YLZD group, medium-dose YLZD group, and high-dose YLZD group, with five mice in each group. The NAFLD mouse model was induced with high-fat feed. After successful modeling, the drug group was given corresponding concentrations of drugs by gavage. In contrast, the control and model groups were given equal amounts of distilled water by gavage once a day for four weeks.The pathological changes in the liver tissue of mice after different drug treatments were observed. Flow cytometry was used to detect the helper T cell 17(CD3~+CD4~+IL-17~+Th17 cells), regulatory T cell(CD3~+CD4~+Foxp3~+Treg cells) levels, Th17/Treg in mesenteric lymph nodes. RT-qPCR detected liver IL-17 and IL-10.Results:Compared with the control group, the weight of the model group mice significantly increased(P<0.01). Hepatocyte ballooning degeneration, steatosis and inflammatory cell infiltration were obvious. After four weeks of drug treatment, compared with the model group, the weight of mice in each dose group of YLZD significantly decreased(P<0.05, P<0.01). Ballooning degeneration, lipoid degeneration, and inflammatory cell infiltration in liver tissue were significantly reduced. In terms of mechanism, the proportion of Th17 in mouse mesenteric lymph nodes of the model group mice significantly reduced(P<0.01), and the mRNA expression levels of IL-17 significantly increased(P<0.05), the Th17/Treg significantly increased(P<0.01),while the proportion of Treg cells significantly decreased(P<0.05), the mRNA levels of IL-10 decreased(P<0.01). However, the proportion of Th17 in mouse mesenteric lymph nodes of YLZD significantly decreased(P<0.01) and the mRNA levels of IL-17 were significantly reduced(P<0.01); while the proportion of Treg cells significantly increased(P<0.01) and the mRNA expression level of IL-10 increased(P<0.01), the Th17/Treg significantly decreased(P<0.01). Conclusion: Yinchen Linggui Zhugan Decoction can significantly alleviate the steatosis and inflammatory cell infiltration of NAFLD mice induced by a high-fat diet, which may exert therapeutic effects by regulating Th17/Treg cell immune balance.更多还原