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黛矾散干预Th17/Treg平衡防治原发性胆汁性胆管炎的作用机制
Mechanism of action of Daifan powder in the prevention and treatment of primary biliary cholangitis via its interventional effect on Th17/Treg balance
文章发布日期:2018年09月30日  来源:  作者:占凯,徐燕,韩梦玲,等  点击次数:253次  下载次数:21次

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【摘要】:目的观察不同剂量的黛矾散对聚肌胞苷酸(Poly I∶C)诱导的原发性胆汁性胆管炎(PBC)模型小鼠外周血辅助性T淋巴细胞17(Th17)、CD4+CD25+Foxp3+调节性T淋巴细胞(Treg)及Th17/Treg比值的影响,进一步探讨黛矾散治疗PBC的作用机制。方法将90只C57BL/6雌性小鼠随机分成正常组、模型组、黛矾散低剂量组、黛矾散中剂量组、黛矾散高剂量组、熊去氧胆酸(UDCA)组,用Poly I:C诱导PBC模型,观察黛矾散对PBC模型小鼠血清学的影响。检测外周血中肝功能相关指标,采用流式细胞技术检测外周血Treg、Th17比例,HE染色检测肝组织病理结构。符合正态分布的计量资料2组间比较采用t检验,多组间比较采用单因素方差分析,进一步两两比较采用LSD-t检验;非正态分布的计量资料2组间比较采用两独立样本Mann-Whitney U非参数检验,多组间比较采用多独立样本Kraskal-Wallis H非参数检验,相关分析采用Spearman相关性检验。结果与正常组比较,模型组和UDCA组血清ALT、AST、ALP、TBil、DBil水平均有升高(P值均<0.05),与模型组比较,黛矾散各组血清ALT、AST、ALP、TBil、DBil水平均有下降(P值均<0.05);PBC模型组小鼠外周血Treg、Th17比例与ALT、AST、ALP、TBil、DBil均无相关性(P值均<0.05)。模型组小鼠外周血Th17百分比增高,与正常组小鼠比较,差异有统计学意义(P<0.01);黛矾散各组小鼠外周血Th17百分比下降,与模型组小鼠比较,差异均有统计学意义(P值均<0.01)。模型组小鼠外周血Treg百分比下降,与正常组小鼠比较,差异有统计学意义(P<0.01);黛矾散各治疗组小鼠外周血Treg百分比下降,与正常组小鼠比较,差异均有统计学意义(P值均<001)。模型组小鼠外周血Th17/Treg比值增高,与正常组小鼠比较,差异有统计学意义(P<0.01);黛矾散各治疗组小鼠外周血Th17/Treg比值下降,与正常组小鼠比较,差异均有统计学意义(P值均<0.01)。结论黛矾散通过打破Th17/Treg平衡,降低免疫炎症反应发挥治疗PBC的作用。
【Abstract】:ObjectiveTo investigate the effect of different doses of Daifan powder on peripheral blood T helper 17 (Th17) cells, CD4+CD25+Foxp3+ regulatory T (Treg) cells, and Th17/Treg ratio in a mouse model of primary biliary cholangitis (PBC) induced by polyinosinic-polycytidylic acid (Poly I∶C) and the mechanism of action of Daifan powder in the treatment of PBC. MethodsA total of 90 C57BL/6 female mice were randomly divided into normal group, model group, low-, middle-, and high-dose Daifan powder groups, and ursodeoxycholic acid (UDCA) group. Poly I:C was used to establish a model of PBC, and the effect of Daifan powder on the serology of PBC mice was observed. Liver function parameters were measured; flow cytometry was used to measure the percentages of peripheral blood Treg cells and Th17 cells; HE staining was used to observe liver pathology. The t-test was used for comparison of normally distributed continuous data between two groups; a one-way analysis of variance was used for comparison between multiple groups, and the least significant difference t-test was used for further comparison between two groups. The Mann-Whitney U test (for two independent samples) was used for comparison of non-normally distributed continuous data between two groups; the Kruskal-Wallis H test (for two or more independent samples) was used for comparison between multiple groups. Spearman correlation was used for the correlation analysis. ResultsCompared with the normal group, the model group and the UDCA group had significant increases in the serum levels of alanine aminotransferase (ALT), aspartate aminotransferase (AST), alkaline phosphatase (ALP), total bilirubin (TBil), and direct bilirubin (DBil) (all P<005). Compared with the model group, the low-, middle-, and high-dose Daifan powder groups had significant reductions in the serum levels of ALT, AST, ALP, TBil, and DBil (all P<0.05). In the model group, peripheral blood Th17/Treg ratio was not correlated with ALT, AST, ALP, TBil, and DBil (all P<0.05). Compared with the normal group, the model group had a significant increase in the percentage of peripheral blood Th17 cells (P<0.01); compared with the model group, the low-, middle-, and high-dose Daifan powder groups had a significant reduction in the percentage of peripheral blood Th17 cells (all P<0.01). Compared with the normal group, the model group had a significant reduction in the percentage of peripheral blood Treg cells (P<0.01); compared with the model group, the low-, middle-, and high-dose Daifan powder groups had a significant increase in the percentage of peripheral blood Treg cells (all P<0.01). Compared with the normal group, the model group had a significant increase in peripheral blood Th17/Treg ratio (P<0.01); compared with the model group, the low-, middle-, and high-dose Daifan powder groups had a significant reduction in peripheral blood Th17/Treg ratio (all P<0.01). ConclusionDaifan powder exerts a therapeutic effect on PBC by breaking Th17/Treg balance and reducing immune inflammatory reaction.
【关键字】:黛矾散; 肝硬化, 胆汁性; T淋巴细胞, 辅助诱导; T淋巴细胞, 调节性
【Key words】:Daifansan; liver cirrhosis, biliary; T-lymphocytes, helper-inducer; T-lymphocytes, regulatory
【引证本文】:ZHAN K, XU Y, HAN ML, et al. Mechanism of action of Daifan powder in the prevention and treatment of primary biliary cholangitis via its interventional effect on Th17/Treg balance[J]. J Clin Hepatol, 2018, 34(11): 2373-2378. (in Chinese) 占凯, 徐燕, 韩梦玲, 等. 黛矾散干预Th17/Treg平衡防治原发性胆汁性胆管炎的作用机制[J]. 临床肝胆病杂志, 2018, 34(11): 2373-2378.

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