柴胡疏肝散对HBV感染小鼠Th17/Treg细胞亚群失衡的逆转作用

Effect of Chaihu Shugan powder on reversing imbalance of Th17/Treg cell subgroup in mice with HBV infection

目的探讨柴胡疏肝散对乙型肝炎病毒(HBV)感染小鼠辅助性T细胞/调节性T细胞(Th17/Treg)亚群失衡的逆转作用。方法选取C57BL小鼠35只,随机分成5组,分别设为模型组、低剂量治疗组、中剂量治疗组、高剂量治疗组和对照组,各7只。通过尾静脉将rAAV8-1.3HBV重组病毒注入小鼠体内;治疗组小鼠进行柴胡疏肝散水煎液灌胃治疗,观察小鼠健康状况与精神状态,进行外周血细胞分类计数,肝脏组织病理切片,检测肝脏组织中的免疫球蛋白表达水平、HBV-DNA复制水平、HBV病毒抗原乙肝核心抗原(HBcAg)水平,计算HBcAg阳性细胞比例,流式细胞仪检测Th17/Treg细胞亚群水平。结果对照组小鼠精神状态正常,模型组、治疗组小鼠出现不同程度异常,且体质量与对照组之间具有统计学差异;与对照组相比,模型组炎症细胞水平最高,治疗组从低剂量到高剂量逐渐降低(P<0.05);肝脏组织病理观察,模型组与治疗组肝脏组织出现不同时期病变;各组肝脏指数具有统计学差异;模型组与治疗组小鼠外周血免疫球蛋白(Ig)G、IgM蛋白表达水平高于对照组(P<0.05);模型组、不同剂量治疗组小鼠肝脏组织中HBV-DNA、HBcAg水平依次降低,对照组HBcAg检测为阴性;模型组、治疗组小鼠肝脏组织Th17/Treg细胞亚群比例高于对照组(P<0.05)。结论柴胡疏肝散对HBV感染小鼠Th17/Treg细胞亚群失衡具有逆转作用。

OBJECTIVE To explore the effect of Chaihu Shugan Powder on reversing the imbalance of helper T cells/regulatory T cells(Th17/Treg)cell subsets in mice infected with hepatitis B virus(HBV).METHODS Totally 35 C57 BL mice were enrolled in the study and randomly divided into the model group,the low-dose group,the medium-dose group,the high-dose group and the control group,with 7 mice in each group.The rAAV8-1.3 HBV recombinant virus was injected into the mice through the tail vein;the treatment groups were given gavage with Chaihu Shugansan water decoction.The health and mental status of the mice were observed,the peripheral blood cells were counted separately,the histopathological section of liver tissues was carried out,the expression level of immunoglobins,HBV-DNA replication level and hepatitis B core antigen(HBcAg)level in the liver tissues were detected,the percentages of HBcAg-positive cells were calculated,and the levels of Th17/Treg subsets were detected by using flow cytometry.RESULTS The mice of the control group showed normal mental status,while the mice of the model group and the treatment groups showed abnormal mental status in various degree,and there was significant difference in the body weight between the model group,the treatment groups and the control group.As compared with the control group,the levels of inflammatory cells of the model group were the highest,and the levels of inflammatory cells of the treatment groups showed a downward trend form the low dose to the high dose(P<0.05).The pathological examination of liver tissues showed that the liver tissues of the model group and the treatment group had different stages of lesions,there were significant differences in the liver indexes among the groups;the expression levels of peripheral blood immunoglobulin(Ig)G and IgM of the model group and the treatment groups were significantly higher than those of the control group(P<0.05).The levels of HBV-DNA and HBcAg in liver tissues showed successive downward trends in the model group and the different doses groups,HBcAg was tested negative in the control group;the percentages of Th17/Treg cells subsets in liver tissues were significantly higher in the model group and the treatment groups than in the control group(P<0.05).CONCLUSION Chaihu Shugan powder can reverse the imbalance of Th17/Treg cell subsets in the mice with HBV infection.