大黄虫丸对大鼠原代肝星状细胞BAMBI表达的影响  

Influence of Dahuang Zhechong Pill on expression of BAMBI in primary hepatic stellate cells in rats

作　　者：徐新杰[1] 刘旭东[1] 赵壮志[1] 吕萍[1] Xu Xinjie;Liu Xudong;Zhao Zhuangzhi;Lyu Ping(Department of Liver Disease,Ruikang Hospital,Guangxi University of Chinese Medicine,Guangxi 530011,China)

机构地区：[1]广西中医药大学附属瑞康医院肝病科,广西530011

出　　处：《北京中医药大学学报》2018年第7期572-578,共7页Journal of Beijing University of Traditional Chinese Medicine

基　　金：国家自然科学基金资助项目(No.81473532);广西自然科学基金资助项目(No.2013GXNSFBA019188);广西八桂学者建设专项经费资助项目~~

摘　　要：目的通过检测大鼠原代肝星状细胞(HSC)中骨成形蛋白-激活素膜结合阻断因子(BAMBI)的表达,探讨大黄虫丸抑制及逆转肝纤维化的作用机制。方法采用清洁级Wistar大鼠,成功复制大鼠肝纤维化模型,分离培养在体大鼠原代HSC,分为正常组、模型组和大黄虫丸组。培养24 h后,采用实时荧光定量PCR法检测原代HSC细胞Toll样受体4(TLR4)、髓样分化因子88(My D88)、BAMBI的基因表达变化,采用免疫组化、蛋白质印迹法(Western blot)检测原代HSC中蛋白的表达变化,采用凝胶电泳迁移率实验(EMSA)检测各组细胞核因子κB(NF-κB)的活性。结果在大鼠原代HSC中,与正常组比较,大黄虫丸组和模型组TLR4、My D88蛋白及基因的表达增强(P<0.05、P<0.001),BAMBI蛋白及基因表达降低(P<0.001);与模型组比较,大黄虫丸组TLR4、My D88蛋白及基因的表达降低(P<0.05),BAMBI蛋白及基因表达增强(P<0.05)。EMSA结果显示:与正常组比较,模型组HSC核转录因子NF-κB的活化增强;与模型组比较,大黄虫丸组NF-κB的活化受抑制。结论大黄虫丸可以改善肝纤维化。其作用机制可能是降低肝星状细胞TLR4、My D88的表达,抑制HSC核转录因子NF-κB的活化,进而增加BAMBI的表达,降低HSC对转化生长因子-β(TGF-β)的敏感性,从而改善肝纤维化。Objective To investigate the effective mechanism of Dahuang Zhechong （Rhubarb and Dormant Insect） Pill in inhibition and reversion of hepatic fibrosis through detecting the expression of bone morphogenic protein and activin membrane-bound inhibitor （BAMBI） in primary hepatic stellate cells （HSC）. Methods The model of hepatic fibrosis was established in clean Wistar rats, primary HSC in vivo were isolated and cultured, and then divided into normal group, model group and Dahuang Zhechong Pill group. After culturing for 24 h, the genetic expressions of TLR4, MyD88 and BAMBI in HSC were detected by using real-time fluorescence quantitative polymerase chain reaction （RT-PCR）. The protein expressions of HSC were detected by using immunohistochemistry technique and Western blotting assay, and activity of nuclear factor-κB （NF-κB） was detected by using electrophoretic mobility shift assay （EMSA）. Results Compared with normal group, the protein and genetic expressions of TLR4 and MyD88 increased （P 〈0.001 ）, and protein and genetic expressions of BAMBI decreased （P 〈0. 001 ） in model group. Compared with model group, the protein and genetic expressions of TLR4 and MyD88 decreased （P 〈 0.05 ） , and protein and genetic expressions of BAMBI increased （ P 〈 0.05 ） in Dahuang Zhechong Pill group. The protein and genetic expressions of TLR4 and MyD88 were higher （P 〈 0.05 ） , and protein and genetic expression of BAMBI were lower （ P 〈 0.05 ） in Dahuang Zhechong Pill group than those in normal group. The results of EMSA showed that the activation of NF-κB was improved in model group compared with normal group, and was inhibited in Dahuang Zhechong Pill group compared with model group. Conclusion Dahuang Zhechong Pill can relieve hepatic fibrosis, and the effective mechanism may be related to reducing expressions of TLR4 and MyD88, inhibiting activation of NF-s：B and improving expression of BAMBI in HSC, and reducing HSC sensitivity to TGF-β.

关 键 词：大黄[庶虫]虫丸 原代肝星状细胞 肝纤维化 大鼠 

分 类 号：R285.5[医药卫生—中药学]