大黄■虫丸含药血清调控大鼠原代肝星状细胞BAMBI表达的研究  

Serum Containing Dahuang Zhechong Pills Regulates BAMBI Expression in Rats Primary Hepatic Stellate Cells

作　　者：吕萍[1] 刘旭东[1] 徐新杰[1] 赵壮志[1] LYU Ping;LIU Xudong;XU Xinjie;ZHAO Zhuangzhi(Department of Liver Disease, Ruikang Hospital Affiliated to Guangxi University of Traditional Chinese Medicine, Nanning 530011 , Guangxi, China)

机构地区：[1]广西中医药大学附属瑞康医院肝病科,广西南宁530011

出　　处：《中华中医药学刊》2019年第5期1088-1091,I0008,共5页Chinese Archives of Traditional Chinese Medicine

基　　金：国家自然科学基金项目(81473532);广西自然科学基金项目(2013GXNSFBA019188);广西一流学科建设重点项目(2018XK084);广西八桂学者建设专项经费项目

摘　　要：目的:研究大黄■虫丸含药血清对LPS诱导大鼠肝星状细胞(HSC)骨成形蛋白-激活素膜结合阻断因子(BAMBI)表达的影响及其作用机制。方法:体外培养并鉴定Wistar大鼠原代HSC,提取并制备大黄■虫丸含药血清,分别予空白对照血清、大黄■虫丸中药含药血清培养细胞,LPS刺激后,应用Western Blot法和免疫组化检测HSC中TLR4、MyD88、BAMBI蛋白变化;EMSA检测NF-κB结合蛋白的活性;结果:Western Blot法和免疫组化结果:与空白对照组比较,LPS处理组TLR4、MYD88表达升高,BAMBI表达降低。与LPS处理组比较,含药血清和LPS共处理组TLR4、MYD88降低,BAMBI表达升高。EMSA结果显示,LPS处理组NF-κB结合蛋白的活性增强,LPS与含药血清共处理组NF-κB结合蛋白的活性降低。结论:大黄■虫丸含药血清可通过抑制LPS诱导的HSCs中TLR4-MYD88的表达,进而减少NF-κB活化,促进TGF-β假受体BAMBI的表达,这可能是发挥其抗纤维化作用的机制之一。Objective: To investigate the effect of serum containing Dahuang Zhechong Pills(DHZC) on the expression of bone morphogenetic protein-activator membrane-blocking factor(BAMBI) induced by LPS in rats hepatic stellate cells(HSC) and its mechanism. Methods: The primary HSCs of Wistar rats were cultured and identified in vitro, and the DHZC containing serum was extracted and prepared. The cells were cultured with control serum and DHZC containing serum respectively. After LPS stimulation, Western blot and immunohistochemistry were used to detect TLR4, MyD88 and BAMBI protein in HSCs. EMSA was used to detect NF-κB binding protein activity. Results: Western blot and immunohistochemistry results: Compared with the control group, the expressions of TLR4 and MYD88 wereincreased and the expression of BAMBI was decreased in the LPS treatment group. Compared with the LPS treatment group, TLR4 and MYD88 were decreased and BAMBI expression was increased in the DHZC+LPS group. EMSA results showed that the activity of NF-κB binding protein was increased in LPS treatment group, and the activity of NF-κB binding protein was decreased in DHZC+LPS group. Conclusion: Serum containing Dahuang Zhechong Pills can inhibit the expression of TLR4-MYD88 in LPS-induced HSCs, and further reduce NF-κB activation and promote the expression of TGF-β pseudo-receptor BAMBI, which may be one of the mechanisms of its anti-fibrosis.

关 键 词：BAMBI 大黄[庶虫]虫丸 TLR4通路 HSC 

分 类 号：R285.5[医药卫生—中药学]