藤茶总黄酮调控TGF-β1/Smad信号通路抗肝纤维化机制的研究
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摘要
目的探讨藤茶总黄酮(TF)通过调控转化生长因子β1(TGF-β1)/Smad信号通路抗小鼠肝纤维化的机制。方法将98只昆明小鼠(雌雄兼用)分为对照组、模型组、秋水仙碱组(0.2mg/kg),TF高、中、低剂量组(300、150、75mg/kg),其中模型组28只(因实验过程中会有死亡),其余各组每组14只。除对照组外,给予各组小鼠背部皮下注射25%四氯化碳(CCl4)花生油溶液,每3天1次,连续10周,造成小鼠肝纤维化模型,对照组背部皮下注射等量100%花生油溶液(每3天1次,连续10周);造模同时,除对照组和模型组小鼠给予蒸馏水灌胃外,其余各组小鼠给予相应药物灌胃给药,每天1次,连续10周。第10周末,摘眼球取血,处死小鼠取肝组织。生化分析法检测各组小鼠血清丙氨酸氨基转移酶(ALT)及天冬氨酸氨基转移酶(AST)水平。ELISA法检测肝组织中Ⅰ型胶原蛋白(ColⅠ)、Ⅲ型胶原蛋白(ColⅢ)及TGF-β1的水平。制作肝组织切片HE染色,免疫组织化学法检测肝组织TGF-β1/Smad信号通路中主要蛋白的表达。结果与模型组比较,TF高、中、低剂量组肝纤维化小鼠血清ALT、AST水平和肝组织ColⅠ、ColⅢ水平明显降低(P<0.05);给药各组小鼠肝组织炎症、坏死现象明显减轻,胶原蛋白增生情况改善。结论 TF可通过调节TGF-β1/Smad信号通路中主要蛋白的表达,从而达到抗肝纤维化的作用。
Objective To explore the mechanism of tengcha flavonoids(TF)on anti-hepatic fibrosis by regulating TGF-β1/Smad signaling pathway.Methods 98 Kunming mice(including male and female)were divided into the control group,the model group,the colchicine group(0.2 mg/kg),the TF high-,middle-,lowdose groups(300,150,75 mg/kg)with 14 mice in each group except 28 mice in the model group.Mice were given hypodermic 25% CCl4 peanut oil once every 3 dfor 10 weeks to establish hepatic fibrosis models except for the control group.At the same time,mice were given distilled water by intragastric administration in the control group and the model group,and corresponding drugs once a day for 10 weeks in other groups.At the end of week 10,the levels of ALT,AST in serum were detected by biochemical analysis.CollagenⅠ(ColⅠ),CollagenⅢ(ColⅢ)in liver were detected by ELISA method.The expression of the major proteins in TGF-β1/Smad signaling pathway in liver tissues was determined by immunohistochemistry.Results Comparing with the model group,TF can obviously reduce the serum ALT,AST and ColⅠ,ColⅢin liver tissue in mice with liver fibrosis(P<0.05).The inflammation and necrosis of liver tissue significantly reduced,and collagen hyperplasia improved in the TF group.Conclusion TF plays an anti-fibrosis role by regulating the expression of the major proteins in TGF-β1/Smad signaling pathway.
Objective To explore the mechanism of tengcha flavonoids(TF)on anti-hepatic fibrosis by regulating TGF-β1/Smad signaling pathway.Methods 98 Kunming mice(including male and female)were divided into the control group,the model group,the colchicine group(0.2 mg/kg),the TF high-,middle-,lowdose groups(300,150,75 mg/kg)with 14 mice in each group except 28 mice in the model group.Mice were given hypodermic 25% CCl4 peanut oil once every 3 dfor 10 weeks to establish hepatic fibrosis models except for the control group.At the same time,mice were given distilled water by intragastric administration in the control group and the model group,and corresponding drugs once a day for 10 weeks in other groups.At the end of week 10,the levels of ALT,AST in serum were detected by biochemical analysis.CollagenⅠ(ColⅠ),CollagenⅢ(ColⅢ)in liver were detected by ELISA method.The expression of the major proteins in TGF-β1/Smad signaling pathway in liver tissues was determined by immunohistochemistry.Results Comparing with the model group,TF can obviously reduce the serum ALT,AST and ColⅠ,ColⅢin liver tissue in mice with liver fibrosis(P<0.05).The inflammation and necrosis of liver tissue significantly reduced,and collagen hyperplasia improved in the TF group.Conclusion TF plays an anti-fibrosis role by regulating the expression of the major proteins in TGF-β1/Smad signaling pathway.
引文
[1]冯涵,李娜,王华林,等.藤茶中黄酮类成分的功效研究进展[J].公共卫生与预防医学,2018,29(1):82-86.
[2]李刚,郑作文,唐云丽.藤茶总黄酮体外抗人肝癌细胞作用研究[J].中国药房,2008,19(9):652-654.
[3]欧贤红,吕林艳,郑作文.藤茶提取物抗慢性肝纤维化作用[J].中国实验方剂学杂志,2011,17(3):132-134.
[4]邝满元.藤茶总黄酮对大鼠肝纤维化的防治作用[J].现代生物医学进展,2008,8(12):2445-2447.
[5]邝满元,罗明英,贾蕾.藤茶总黄酮对实验性肝纤维化大鼠TGF-β_1表达的影响[J].中国民族民间医药,2009,18(6):6-8.
[6]BAGHERNIYA M,NOBILI V,BLESSO C N,et al.Medicinal plants and bioactive natural compounds in the treatment of non-alcoholic fatty liver disease:a clinical review[J].Pharmacol Res,2018,130(6):213-240.
[7]XU F,LIU C,ZHOU D,et al.TGF/SMAD pathways and its regulation in hepatic fibrosis[J].J Histochem Cytochem,2016,64(3):157-167.
[8]魏伟,吴希美,李元建.药理实验方法学[M].4版.北京:人民卫生出版社,2010:1135.
[9]李洁,邱建利,许华.加味茵陈四逆汤对肝纤维化小鼠Smad7及Ⅰ、Ⅲ型胶原蛋白表达的影响[J].中药材,2016,39(6):1384-1388.
[10]杨小瑜.肝纤维化的发病机制研究进展[J].山东医药,2017,57(11):108-110.
[11]GRESSNER A M,WEISKIRCHEN R.Modern pathogenetic concepts of liver fibrosis suggest stellate cells and TGF-beta as major players and therapeutic targets[J].JCell Mol Med,2006,10(1):76-99.
[12]李欣,戴立里.转化生长因子β1/Smads信号转导途径及其与肝纤维化的关系[J].重庆医学,2007,36(18):1887-1889.
[13]张玉,周曦,易龙,等.二氢杨梅素通过TGF-β1/Smad信号通路抑制肝星状细胞活化的作用研究[J].第三军医大学学报,2018,40(4):282-289.
[14]LIN Y,LUO H,WANG X,et al.Flavanones from sedum sarmentosum bunge alleviate CCl4-induced liver fibrosis in rats by targeting TGF-β1/TβR/Smad pathway in turn Inhibiting epithelial mesenchymal transition[J].Evid Based Complement Alternat Med,2018,2018(8):1-10.
[15]ARGENTOU N,GERMANIDIS G,HYTIROGLOU P,et al.TGF-βsignaling is activated in patients with chronic HBV infection and repressed by SMAD7overexpression after successful antiviral treatment[J].Inflamm Res,2016,65(5):355-365.
[16]左魁阳,陈梦茜,孟娜娜,等.TGF-β与器官纤维化的研究进展[J].中国当代医药,2017,24(17):12-16.
[2]李刚,郑作文,唐云丽.藤茶总黄酮体外抗人肝癌细胞作用研究[J].中国药房,2008,19(9):652-654.
[3]欧贤红,吕林艳,郑作文.藤茶提取物抗慢性肝纤维化作用[J].中国实验方剂学杂志,2011,17(3):132-134.
[4]邝满元.藤茶总黄酮对大鼠肝纤维化的防治作用[J].现代生物医学进展,2008,8(12):2445-2447.
[5]邝满元,罗明英,贾蕾.藤茶总黄酮对实验性肝纤维化大鼠TGF-β_1表达的影响[J].中国民族民间医药,2009,18(6):6-8.
[6]BAGHERNIYA M,NOBILI V,BLESSO C N,et al.Medicinal plants and bioactive natural compounds in the treatment of non-alcoholic fatty liver disease:a clinical review[J].Pharmacol Res,2018,130(6):213-240.
[7]XU F,LIU C,ZHOU D,et al.TGF/SMAD pathways and its regulation in hepatic fibrosis[J].J Histochem Cytochem,2016,64(3):157-167.
[8]魏伟,吴希美,李元建.药理实验方法学[M].4版.北京:人民卫生出版社,2010:1135.
[9]李洁,邱建利,许华.加味茵陈四逆汤对肝纤维化小鼠Smad7及Ⅰ、Ⅲ型胶原蛋白表达的影响[J].中药材,2016,39(6):1384-1388.
[10]杨小瑜.肝纤维化的发病机制研究进展[J].山东医药,2017,57(11):108-110.
[11]GRESSNER A M,WEISKIRCHEN R.Modern pathogenetic concepts of liver fibrosis suggest stellate cells and TGF-beta as major players and therapeutic targets[J].JCell Mol Med,2006,10(1):76-99.
[12]李欣,戴立里.转化生长因子β1/Smads信号转导途径及其与肝纤维化的关系[J].重庆医学,2007,36(18):1887-1889.
[13]张玉,周曦,易龙,等.二氢杨梅素通过TGF-β1/Smad信号通路抑制肝星状细胞活化的作用研究[J].第三军医大学学报,2018,40(4):282-289.
[14]LIN Y,LUO H,WANG X,et al.Flavanones from sedum sarmentosum bunge alleviate CCl4-induced liver fibrosis in rats by targeting TGF-β1/TβR/Smad pathway in turn Inhibiting epithelial mesenchymal transition[J].Evid Based Complement Alternat Med,2018,2018(8):1-10.
[15]ARGENTOU N,GERMANIDIS G,HYTIROGLOU P,et al.TGF-βsignaling is activated in patients with chronic HBV infection and repressed by SMAD7overexpression after successful antiviral treatment[J].Inflamm Res,2016,65(5):355-365.
[16]左魁阳,陈梦茜,孟娜娜,等.TGF-β与器官纤维化的研究进展[J].中国当代医药,2017,24(17):12-16.