基于TGF-β/Smad信号通路探讨牛大力总黄酮对四氯化碳致肝纤维化大鼠的保护作用  

Protective effect and mechanism of total flavonoids of Millettia speciosa Champ.on carbon tetrachloride induced liver fibrosis in rats

作　　者：陈秋贵 覃妮 陆世银 张鑫 侯玲 邓朝旺 张琳华 韦玮琴 CHEN Qiugui;QIN Ni;LU Shiyin;ZHANG Xin;HOU Ling;DENG Chaowang;ZHANG Linhua;WEI Weiqin(Liuzhou Hospital of Traditional Chinese Medicine,Liuzhou 545026,Guangxi,China;Liuzhou Municipal Liutie Central Hospital,Liuzhou 545000,Guangxi,China)

机构地区：[1]柳州市中医医院(柳州市壮医医院),广西柳州545026 [2]柳州市柳铁中心医院,广西柳州545000

出　　处：《现代中西医结合杂志》2025年第3期299-305,共7页Modern Journal of Integrated Traditional Chinese and Western Medicine

基　　金：柳州市科技计划项目(2020NBAA0802);广西中医药大学自然科学研究面上项目(2020MS055)。

摘　　要：目的观察牛大力总黄酮对四氯化碳(CCl_(4))致肝纤维化大鼠肝功能、肝纤维化的影响,并基于氧化应激、炎症反应、转化生长因子-β(TGF-β)/Smad信号通路探讨其可能作用机制。方法取雄性SD大鼠33只,随机选择其中6只作为正常组,其余大鼠采用腹腔注射40%CCl_(4)橄榄油溶液(0.1 mL/100 g)方法复制肝纤维化模型,2次/周,连续注射12周。造模第8周末将24只肝纤维化建模成功大鼠随机分为模型组、秋水仙碱组、牛大力总黄酮低剂量组、牛大力总黄酮高剂量组,每组6只。从造模第9周开始,秋水仙碱组大鼠每天给予0.2 mg/kg秋水仙碱灌胃,牛大力总黄酮低、高剂量组大鼠分别每天给予5 mg/kg、20 mg/kg牛大力总黄酮溶液灌胃,正常组与模型组大鼠每天灌胃等体积的生理盐水,均连续灌胃4周。取血检测肝功能指标[丙氨酸转氨酶(ALT)、天冬氨酸转氨酶(AST)]、肝纤维化指标[透明质酸(HA)、层黏连蛋白(LN)、Ⅲ型前胶原(PCⅢ)、Ⅳ型胶原(Ⅳ-C)]、氧化应激指标[丙二醛(MDA)、超氧化物歧化酶(SOD)]、炎症因子[白细胞介素-1β(IL-1β)、白细胞介素-8(IL-8)、白细胞介素-6(IL-6)]水平;HE染色、Masson染色及天狼星红染色观察肝组织病理学形态;Western blot法检测肝组织中TGF-β_(1)、Smad3、Smad7蛋白表达情况。结果与正常组比较,模型组大鼠血清ALT、AST、HA、LN、PCⅢ、Ⅳ-C、MDA、IL-1β、IL-8、IL-6水平和肝组织中TGF-β_(1)、Smad3蛋白相对表达量均明显升高(P均<0.05),血清SOD水平和肝组织中Smad7蛋白相对表达量均明显降低(P均<0.05),肝组织出现明显纤维化;与模型组比较,各药物组大鼠血清ALT、AST、LN、HA、PCⅢ、Ⅳ-C、MDA、IL-1β、IL-8、IL-6水平和牛大力总黄酮低、高剂量组大鼠肝组织中TGF-β_(1)、Smad3蛋白相对表达量均明显降低(P均<0.05),各药物组大鼠血清SOD水平和牛大力总黄酮低、高剂量组大鼠肝组织中Smad7蛋白相对表达量Objective It is to observe the effect of total flavones from Millettia speciosa Champ.(TFMC)on liver function,liver fibrosis of rats with carbon tetrachloride(CCl_(4))-induced liver fibrosis,and to explore its possible mechanisms of action based on oxidative stress,inflammatory response,and transforming growth factor-β(TGF-β)/Smad signaling pathway.Methods Thirty-three male SD rats were taken,6 of which were randomly selected as normal group,and the other rats were injected intraperitoneally with 40%CCl_(4)olive oil solution(0.1 mL/100 g)to replicate models of hepatic fibrosis,twice/week,continuously treated for 12 weeks.At the end of the 8th week of modeling,24 successfully modeled rats with hepatic fibrosis were randomly divided into model group,colchicine group,the low dose and high-dose groups of TFMC,with 6 rats in each group.Starting from the 9th week of modeling,the rats in the colchicine group were given colchicine 0.2 mg/kg by gavage every day,the rats in the low dose and high dose groups of TFMC were respectively given TFMC 5 mg/kg and 20 mg/kg by gavage every day,and the rats in the normal group and model group were given the same volume of saline by gavage every day,all continuously gavaged for 4 weeks.The levels of liver function indexes[alanine aminotransferase(ALT),aspartate aminotransferase(AST)],hepatic fibrosis indexes[hyaluronic acid(HA),laminin(LN),pre-collagen typeⅢ(PCⅢ),collagen typeⅣ(ⅣC)],oxidative stress indexes[malondialdehyde(MDA),superoxide dismutase(SOD)],inflammatory factors[interleukin-1β(IL-1β),interleukin-8(IL-8),and interleukin-6(IL-6)]were detected;the pathologic morphology of liver tissues was observed by HE staining,Masson staining and Sirius red staining;the protein expressions of TGF-β_(1),Smad3,and Smad 7 in liver tissues were detected by Western blot.Results Compared with the normal group,the serum levels of ALT,AST,HA,LN,PCⅢ、Ⅳ-C,MDA,IL-1β,IL-8,IL-6 and the relative protein expressions of TGF-β_(1)and Smad3 in liver tissues of rats in the model group we

关 键 词：牛大力 总黄酮 肝纤维化 抗氧化 炎症因子 转化生长因子-β/Smad通路 

分 类 号：R-332[医药卫生]