鳖甲煎丸抑制肝星状细胞激活减轻肝纤维化的作用机制  

The mechanism of Biejiajian pill in inhibiting hepatic stellate cell activation and alleviating liver fibrosis

作　　者：左艳洁[1] 李鑫 崔佳林 张荣花 李彩霞[1] 崔立华[1] 章广玲 张淑坤[1] 兰涛 ZUO Yan-jie;LI Xin;CUI Jia-lin(Tianjin Integrated Traditional Chinese and Western Medicine Hospital,Tianjin Nankai Hospital(300100),China)

机构地区：[1]天津市中西医结合医院,天津市南开医院,天津300100 [2]河北省沧州市人民医院肝胆胰外科,沧州061000 [3]天津医科大学研究生院,天津300700 [4]华北理工大学临床医学院,河北省医工融合精准医疗重点实验室,唐山063000

出　　处：《中国中西医结合外科杂志》2024年第4期557-561,共5页Chinese Journal of Surgery of Integrated Traditional and Western Medicine

基　　金：河北省创新能力提升计划——高水平人才团队建设专项(205A7701H)。

摘　　要：目的:研究鳖甲煎丸调节AMP依赖的蛋白激酶(AMPK)、抑制肝星状细胞(HSC)激活和肝纤维化的作用机制。方法:将24只健康雄性SPF级C57BL/6小鼠随机分为对照、肝纤维化模型和鳖甲煎丸治疗组,每组8只。采用高脂高糖饮食联合腹腔注射四氯化碳制备小鼠肝纤维化模型。自第6周始,鳖甲煎丸治疗组给予鳖甲煎丸(1.365 g/kg)灌胃,1次/d,至第12周检测血清三酰甘油(TG)、胆固醇(TC)、高密度脂蛋白(HDL)、低密度脂蛋白(LDL)、谷丙转氨酶(ALT)、谷草转氨酶(AST)和碱性磷酸酶(ALP)水平,取肝组织行HE染色、天狼星红染色和α-平滑肌肌动蛋白(α-SMA)免疫组织化学染色,分别观察肝组织病理学改变、胶原纤维增生和HSC活化状态,并用Western blot检测AMPK蛋白表达和活性。结果:与对照组比较,肝纤维化模型组小鼠血清ALT、AST、ALP和LDL升高,TG、TC及HDL降低(P<0.05);与肝纤维化模型组比较,鳖甲煎丸治疗组小鼠血清ALT、AST和ALP降低(P<0.05),而TG、TC、HDL和LDL差异无统计学意义。肝纤维化模型组HE染色可见肝细胞排列紊乱、有严重脂肪变性和部分气球样变,并伴有炎症细胞浸润和点片状坏死灶;天狼星红染色可见明显的纤维组织增生,α-SMA表达阳性;总AMPK和p-AMPK蛋白表达较对照组升高;与肝纤维化模型组比较,鳖甲煎丸治疗组肝病理改变、纤维组织增生和α-SMA阳性表达程度降低,AMPK和p-AMPK蛋白表达均降低(P<0.05)。结论:鳖甲煎丸能抑制HSC激活、减缓肝纤维化进程,其作用机制与抑制AMPK蛋白表达和活性有关。Objective To investigate the mechanism of Biejiajian pill in regulating AMP dependent protein kinase(AMPK),inhibiting hepatic stellate cell activationand liver fibrosis.Methods Healthy male SPF grade C57BL/6 mice were randomly divided into control,liver fibrosis,and Biejiajian pill group(n=8).Mice of the liver fibrosis and Biejiajian pill group were induced liver fibrosis model with a high-fat and high sugar diet combined with intraperitoneal injection of carbon tetrachloride.The Biejiajian pill group was given Biejiajian pill(1.365 g/kg)by gavage once a day,starting from the 6th week.At the 12th week,serum TG,TC,HDL,LDL,ALT,AST,and ALP were detect,and HE staining,Sirius Red staining,andα-smooth muscle actin(α-SMA)immunohistochemical staining were used to observe the pathological changes,collagen fiber proliferation,and hepatic stellate cells(HSC)activation respectively.In addition,western blot was used to detect the expression and activity of AMPK protein.Results The serum ALT,AST,ALP,and LDL of mice in the liver fibrosis group increased,while TG,TC,and HDL decreased compared to the control group(P<0.05).Compared with the liver fibrosis group,there was a significant difference in the decrease of serum ALT,AST,and ALP in the Biejiajian pill group(P<0.05),while there was no statistically significant difference in TG,TC,HDL,and LDL(P>0.05).HE staining showed disordered arrangement of liver cells,severe steatosis,and partial ballooning in the liver fibrosis group,accompanied by inflammatory cell infiltration and patchy necrotic lesions.Sirius red staining shows obvious fibrous tissue proliferation,α-SMA expression is positive.The expression of total AMPK and p-AMPK proteins increased compared to the control group.Compared with the liver fibrosis group,the liver pathological changes,fibrous tissue proliferation,and the positive expression level ofα-SMA were decreased,and both the expression of AMPK and p-AMPK proteins were decreased(P<0.05).Conclusion Biejiajian pill inhibits HSC activation and liver fibrosis,and

关 键 词：AMP依赖的蛋白激酶 肝星状细胞 肝纤维化 鳖甲煎丸 

分 类 号：R735.7[医药卫生—肿瘤] R256.4[医药卫生—临床医学]