苦参素注射液通过TGF-β_(1)/Smads通路减轻糖尿病大鼠肝纤维化的研究  

Oxymatrine injection alleviates hepatic fibrosis in diabetic rats via TGF-β_(1)/Smads pathway

作　　者：姚飞 王丽 YAO Fei;Wang Li(Handan Iron and Steel Hospital,Handan 056001,Hebei,China)

机构地区：[1]邯郸邯钢医院,河北邯郸056001

出　　处：《现代中西医结合杂志》2023年第20期2809-2816,共8页Modern Journal of Integrated Traditional Chinese and Western Medicine

基　　金：河北省医学科学研究课题(20220473)。

摘　　要：目的研究苦参素注射液对糖尿病大鼠肝纤维化的影响,并基于转化生长因子-β_(1)(TGF-β_(1))/Smads通路探索其作用机制。方法随机从55只大鼠中取10只作为正常组,剩余45只大鼠采用高脂高糖饮食喂养4周后腹腔注射链脲佐菌素构建糖尿病模型,将40只成模大鼠随机分为模型组、苦参素注射液组、SB431542组和苦参素注射液+SB431542组,每组10只。苦参素注射液组给予50 mg/kg苦参素注射液腹腔注射,SB431542组给予0.1 mg/kg SB431542腹腔注射,苦参素注射液+SB431542组给予50 mg/kg苦参素注射液和0.1 mg/kg SB431542腹腔注射,均1次/d。连续注射4周后检测空腹血糖(FPG)水平,肝脏超声测量门静脉近段内径(PDPV)、远段内径(DDPV)、血流速度(VPV)、杨氏模量,取肝脏计算肝脏质量指数(LVI),放射免疫法检测肝组织中肝纤4项[透明质酸(HA)、层黏连蛋白(LN)、Ⅲ型前胶原(PCⅢ)、Ⅳ型胶原(CⅣ)]水平,HE染色和天狼星红染色观察肝组织病理学形态和纤维化情况;Western blot法检测肝组织中TGF-β_(1)、Smad3、p-Smad3、Smad7、Ⅰ型胶原蛋白(Col-Ⅰ)、Ⅲ型胶原蛋白(Col-Ⅲ)、基质金属蛋白酶-2(MMP-2)、基质金属蛋白酶-9(MMP-9)蛋白表达情况。结果与模型组比较,各药物组FPG水平、PDPV、DDPV、杨氏模量、LVI及肝组织中HA、LN、PCⅢ、CⅣ水平均明显降低(P均<0.05),苦参素注射液组和苦参素注射液+SB431542组VPV均明显增快(P均<0.05);各药物组肝小叶结构破坏、肝细胞脂肪变性、炎性细胞浸润和汇管区、门静脉周围、肝小叶周围纤维化等病变均明显减轻,胶原容积分数明显降低(P均<0.05),肝组织中TGF-β_(1)、Col-I、Col-Ⅲ蛋白相对表达量及p-Smad3/Smad3比值均明显降低(P均<0.05),肝组织中Smad7、MMP-2、MMP-9蛋白相对表达量均明显升高(P均<0.05)。苦参素注射液组+SB431542组上述各指标改善情况均明显优于苦参素注射液组和SB431542组(P均<0.05)。结论苦参素�Objective It is to investigate the effect of oxymatrine injection(OMTJ)on hepatic fibrosis in diabetic(DM)rats,and explore its mechanism based on transforming growth factor-β_(1)(TGF-β_(1))/Smads pathway.Methods Fifty-five rats were randomly selected,in which 10 rats were selected as the normal group,and the remaining 45 rats were fed with high fat and high sugar diet for 4 weeks and then given streptozotocin by intraperitoneal injection to establish the DM rat models.40 successfully modeled rats were randomly divided into model group,OMTJ group,SB431542 group and OMTJ+SB431542 group,with 10 rats in each group.The OMTJ group was given 50 mg/kg OMTJ by intraperitoneal injection,the SB431542 group was given 0.1 mg/kg SB431542 by intraperitoneal injection,the OMTJ+SB431542 group was given 50 mg/kg OMTJ+0.1 mg/kg SB431542 by intraperitoneal injection,all once daily,continuously treated for 4 weeks.After treatment,the level of fasting plasma glucose(FPG)was detected,the proximal diameter of portal vein(PDPV),distal diameter of portal vein(DDPV),velocity of portal vein(VPV)and Young's modulus were detected by liver ultrasound,the liver mass index(LVI)was calculated,the levels of hyaluronic acid(HA),laminin(LN),typeⅢprocollagen(PCⅢ),typeⅣcollagen(CⅣ)in hepatic tissue were determined by radioimmuno-assay,the histopathological changes and fibrosis of hepatic tissue were observed by HE staining or sirius red staining,the expressions of TGF-β_(1),Smad3,p-Smad3,Smad7,type I Collagen(Col-Ⅰ),Col-Ⅲ,matrix metalloproteinase 2(MMP2)and matrix metalloproteinase 9(MMP9)were detected by Western blotting.Results Compared with the model group,the FPG level,PDPV,DDPV,VPV,Young's modulus,LVI and the levels of HA,LN,PCⅢ,CⅣin hepatic tissue in each treatment group were significantly decreased(all P<0.05),the VPV in the OMTJ group and OMTJ+SB431542 group was significantly faster(both P<0.05).The lesions such as structural destruction of liver lobules,hepatocellular steatosis,inflammatory cell infiltration and fibrosis in

关 键 词：糖尿病 肝纤维化 苦参素注射液 转化生长因子-β_(1)/Smads通路 

分 类 号：R-332[医药卫生]