基于肠道菌群探讨连翘提取物的抗肝纤维化作用  

Study on anti-liver fibrosis effect of Forsythia suspensa extract based on gut microbiota

作　　者：王成 付珂 龚莉虹 马程 刘艳芳 李芸霞[1] WANG Cheng;FU Ke;GONG Li-hong;MA Cheng;LIU Yan-fang;LI Yun-xia(State Key Laboratory of Southwestern Chinese Medicine Resources,School of Pharmacy,Chengdu University of Traditional Chinese Medicine,Chengdu 611137,China)

机构地区：[1]成都中医药大学药学院,西南特色中药资源国家重点实验室,成都611137

出　　处：《中华中医药杂志》2023年第5期2028-2035,共8页China Journal of Traditional Chinese Medicine and Pharmacy

基　　金：国家自然科学基金面上项目(No.81891012,No.81630101,No.U19A2010);国家中医药管理局中医药创新团队及人才支持计划项目(No.ZYYCXTD-D-202209);四川省科技支撑计划项目(No.2021JDRC0041)。

摘　　要：目的:基于肠道菌群探讨连翘提取物(FSE)对四氯化碳(CCl_4)诱导小鼠肝纤维化的改善作用及其机制。方法:使用CCl_4建立肝纤维化模型,每日灌胃FSE(4 g/kg)进行预防治疗。采用HE染色、Masson染色和天狼星红染色对肝组织进行病理学观察和纤维化评价;利用生化试剂盒测定血清中谷丙转氨酶(ALT)、谷草转氨酶(AST)、碱性磷酸酶(ALP)、γ-谷氨酰转肽酶(γ-GT)、脂多糖(LPS)、巨噬细胞炎症蛋白1(MIP-1)、肿瘤坏死因子α(TNF-α)的水平以及肝组织中羟脯氨酸(HYP)、透明质酸(HAase)、层粘连蛋白(LN)、Ⅳ型胶原(Ⅳ-C)、Ⅲ型前胶原肽(PCⅢ)的含量;运用RT-q PCR法检测肝组织中α-平滑肌肌动蛋白(α-SMA)、Ⅰ型胶原蛋白(CollagenⅠ)、白细胞介素1β(IL-1β)、白细胞介素6(IL-6)、TNF-α m RNA的表达;采用16S r RNA基因测序研究各组小鼠肠道菌群的变化;利用免疫组化和RT-q PCR法检测肠道黏膜屏障关键蛋白紧密连接蛋白1(ZO-1)、闭合蛋白(Occludin)、封密蛋白1(Claudin-1)及其m RNA的表达。结果:FSE可以改善CCl_4引起的肝组织病理损伤、肝功能异常、胶原沉积、炎症和纤维化。此外,16S r RNA基因测序证实了FSE对肠道菌群紊乱的纠正作用。同时,FSE通过促进紧密连接蛋白的表达,恢复了肠上皮屏障功能。结论:FSE通过抑制炎症、调节肠道微生物群稳态以及促进肠黏膜屏障的修复减轻CCl_4诱导的肝纤维化。Objective:To investigate the effects and underlying mechanism of Forsythia suspensa extract(FSE)on carbon tetrachloride(CCl4)-induced liver fibrosis in mice based on gut microbiota.Methods:The liver fibrosis model was established by using CCl4,and FSE(4 g/kg)was orally administered daily for prevention and treatment.Hematoxylin-eosin staining,Masson trichrome staining and Sirius red staining were used to observe liver histopathology and evaluate fibrosis.The contents of ALT,AST,ALP,-GT,LPS,MIP-1 and TNF-αin serum,as well as HYP,HAase,LN,IV-C and PC-Ⅲ in liver tissues were determined by biochemical kits.The mRNA expression ofα-SMA,Collagen Ⅰ,IL-1β,IL-6 and TNF-αin liver tissues was detected by RTqPCR.16S rRNA gene sequencing was used to study the changes of gut microbiota in mice in each group.Immunohistochemistry and RT-qPCR were used to detect the expression of key intestinal mucosal barrier proteins ZO-1,Occludin,Claudin-1 and their genes.Results:FSE could ameliorate CCl-induced liver tissue pathological damage,liver dysfunction,collagen deposition,inflammation and fibrosis.In addition,16S rRNA gene sequencing confirmed the corrective effect of FSE on gut microbiota disorder.At the same time,FSE restored intestinal epithelial barrier function by promoting the expression of tight junction proteins.Conclusion:FSE alleviated CCl-induced liver fibrosis by inhibiting inflammation,regulating gut microbiota homeostasis and promoting intestinal mucosal barrier repair.

关 键 词：连翘提取物 肝纤维化 肠道菌群 肠黏膜屏障 四氯化碳 

分 类 号：R285.5[医药卫生—中药学]