巴芪柔肝方促进Th22细胞活化及IL-22分泌抗肝纤维化机制  

Mechanism of Baqi Rougan Formula in Promoting Activation of Th22 Cells and Secretion of IL-22 against Liver Fibrosis

作　　者：厉姣龙 郭丽坤[2] 周振华[2] LI Jiao-long;GUO Li-kun;ZHOU Zhen-hua(ShuguangSchool of Clinical Medicine,Shanghai University of Traditional Chinese Medicine,Shanghai201203;Department of Hepatology,Shuguang Hospital,Shanghai University of Traditional Chinese Medicine,Shanghai201203)

机构地区：[1]上海中医药大学曙光临床医学院,上海201203 [2]上海中医药大学附属曙光医院肝病科,上海201203

出　　处：《中国中西医结合杂志》2024年第8期944-951,共8页Chinese Journal of Integrated Traditional and Western Medicine

基　　金：国家自然科学基金资助项目(No.81774268);上海市科学技术委员会科技支撑项目(No.21S21900400)。

摘　　要：目的 研究巴芪柔肝方抗肝纤维化的作用机制。方法 60只8周龄的BALB/c小鼠随机分为对照组(Control)、模型组(ConA)和巴芪柔肝方组(RGF)。模型组和RGF组制备刀豆蛋白A(ConA)诱导肝纤维化动物模型,RGF组随后灌服巴芪柔肝方(8.5 g/kg)至实验结束。体外分离健康人外周血CD4+T细胞和树突状细胞(DCs),诱导后分别与人肝星状细胞系(LX2细胞)共培养。检测相关基因和蛋白的表达水平。结果 与对照组比较,模型组小鼠血清谷氨酸丙酮酸转氨酶(ALT)、转化生长因子-β(TGF-β)、透明质酸(HA)、白介素-22(IL-22)、白介素-6(IL-6)、肿瘤坏死因子-α(TNF-α)、白介素-23(IL-23)水平均升高(P<0.01,P<0.05),白蛋白(ALB)水平降低(P<0.01)。与模型组比较,RGF组ALT、TGF-β、HA降低(P<0.01),IL-22、IL-6、TNF-α、IL-23均升高(P<0.01)。病理结果显示巴芪柔肝方对ConA诱导的肝纤维化有保护作用。与空白血清组比较,巴芪柔肝方含药血清组(0.5、1、10μg/mL) DCs分泌的IL-6、TNF-α、IL-23 mRNA水平升高(P<0.01)。在共培养体系中,与空白血清组比较,巴芪柔肝方含药血清组IL-22 mRNA和蛋白的表达水平升高(P<0.01),LX2中α-平滑肌肌动蛋白(α-SMA)和Ⅰ型胶原α1(Col1 α1)的mRNA和蛋白水平降低(P<0.01)。对衰老相关炎症基因的RT-PCR分析显示,巴芪柔肝方含药血清组可以上调基质金属蛋白酶9(MMP9)、IL-6、巨噬细胞炎性蛋白-2(MIP2)、p53和p21 mRNA的表达水平(P<0.01),同时下调基质金属蛋白酶组织抑制因子1(TIMP1) mRNA的表达水平(P<0.01)。结论 巴芪柔肝方通过促进DCs分泌IL-6/TNF-α/IL-23,激活Th22细胞,增加其IL-22的分泌,从而抑制肝星状细胞(HSCs)的活化,促进活化的HSCs衰老。Objective To explore the mechanism of anti-fibrotic action of Baqi Rougan Formula(RGF).Methods Sixty 8-week-old BALB/c mice were randomly assigned to three groups:Control group,Concanavalin A(ConA)model group,and RGF group.Animals model of ConA-induced liver fibrosis were prepared in the model group and RGF group,and the RGF group was then administered with RGF(8.5 g/kg)until the end of the experiment.CD4+T cells and dendritic cells(DCs)were isolated from the peripheral blood of healthy individuals and co-cultured with the human hepatic stellate cell line(LX2)following induction.The expression levels of related genes and proteins were detected.Results Compared to the Control group,the ConA model group exhibited significant increases in serum alanine aminotransferase(ALT),transforming growth factor-beta(TGF-β),hyaluronic acid(HA),interleukin-22(IL-22),interleukin-6(IL-6),tumor necrosis factor-alpha(TNF-α),and interleukin-23(IL-23)levels(P<0.01,P<0.05),along with a decrease in albumin(ALB)level(P<0.01).In contrast,the RGF group showed significant reductions in ALT,TGF-β,HA levels(P<0.01),and increases in IL-22,IL-6,TNF-α,IL-23 levels compared to the model group(P<0.01).Histopathological analysis confirmed the protective effect of the RGF against ConA-induced liver fibrosis.When compared to the blank serum group,the RGF-containing serum groups(0.5,1,10µg/mL)demonstrated elevated mRNA levels of IL-6,TNF-α,IL-23 secreted by DCs(P<0.01).In the co-culture system,compared to the blank serum group,the RGF-containing serum group exhibited increased IL-22 mRNA and protein expression levels(P<0.01),and a decrease in alpha-smooth muscle actin(α-SMA)and Collagen TypeⅠAlpha 1(Col1α1)mRNA and protein levels in LX2 cells(P<0.01).RT-PCR analysis of aging-related inflammatory genes indicated that the RGF-containing serum group could upregulate the expression of matrix metalloproteinase 9(MMP9),IL-6,macrophage inflammatory protein-2(MIP2),p53,and p21 mRNA(P<0.01),while downregulating the expression of tissue inhibitor of m

关 键 词：巴芪柔肝方 树突状细胞 TH22细胞 肝星状细胞 肝纤维化 中药复方 

分 类 号：R285.5[医药卫生—中药学]