矮地茶黄酮抗二甲基亚硝胺诱导大鼠肝纤维化作用及机制研究
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摘要
目的:研究矮地茶黄酮(FAJ)对二甲基亚硝胺(DMN)诱导的大鼠肝纤维化(HF)的保护作用及机制。方法:随机将大鼠分为正常组、模型组、秋水仙碱组(0.1 mg/kg)、FAJ高剂量组(600 mg/kg)、FAJ中剂量组(400 mg/kg)、FAJ低剂量组(200 mg/kg),除正常组外,其余各组腹腔注射0.5%DMN建立大鼠肝纤维化模型,FAJ各剂量组同时给予FAJ干预,1次/d,至实验结束。实验结束后,采集大鼠血液及肝组织标本,检测各组大鼠血清中谷丙转氨酶(ALT))、谷草转氨酶(AST)、超氧化物歧化酶(SOD)和丙二醛(MDA)、透明质酸(HA)、层黏蛋白(LN)水平,免疫组化方法检测各组大鼠肝脏转化生长因子-β1(TGF-β1)表达,观察各组大鼠肝脏组织病理学变化,探讨FAJ对DMN诱导的大鼠肝纤维化的保护作用及机制。结果:与模型组相比,FAJ高剂量组、FAJ中剂量组、FAJ低剂量组大鼠血清中ALT、AST、MDA、HA、LN水平显著降低(P<0.01,P<0.05);FAJ高剂量组、FAJ中剂量组中SOD水平显著升高(P<0.01),FAJ低剂量组SOD虽升高,但无显著性差异(P>0.05);病理切片显示,FAJ干预后肝组织内炎症细胞浸润和纤维化程度明显改善;免疫组化图像显示FAJ各剂量组TGF-β1表达下降。结论:FAJ对DMN诱导的大鼠肝纤维化具有保护作用且呈剂量依赖型,其作用机制可能与抗氧化、抑制肝星状细胞活化有关。
Objective:To study the protective effect and mechanism of flavonoids from Ardisia japonica(Hornsted)BL.(FAJ)on dimethylnitrosamine(DMN)-induced hepatic fibrosis in rats,so as to provide scientific basis for clinical rational drug use and resource development in the future. Methods:Rats were randomly divided into normal group,model group,colchicine group(0.1 mg/kg),FAJ(600 mg/kg,400 mg/kg,200 mg/kg) groups. Except for normal group,each group was intraperitoneally injected with 0.5% DMN to establish hepatic fibrosis model in rats,and FAJ intervention was performed once a day until the end of the experiment. At the end of the experiment,blood and liver tissue samples were collected from the rats. Serum alanine aminotransferase(ALT),aspartate aminotransferase(AST),superoxide dismutase(SOD) and malondialdehyde(MDA),andhyaluronan were detected in each group of rats. The level of acid(HA)and laminin(LN)was detected by immunohistochemistry.The expression of transforming growth factor-β1(TGF-β1)in liver of rats in each group was examined. Histopathological changes of liver in rats of each group were observed by pathology. And protective effect and mechanism of FAJ on DMN-induced hepatic fibrosis in rats was investigated. Results :Compared with the model group,the serum levels of ALT,AST,MDA,HA,and LN in the FAJ(600 mg/kg,400 mg/kg) group were significantly lowed(P <0.01,P <0.05),and the SOD level was significantly increased(P <0.01),the serum levels of ALT,AST,MDA,HA,and LN in the FAJ(200 mg/kg) were significantly lowed(P <0.01,P <0.05),the level of SOD was increased,but there was no significant difference(P >0.05).Pathological sections showed that the degree of inflammatory cell infiltration and fibrosis in liver tissue was significantly improved after FAJ intervention. Immunohistochemical images showed that the expression of TGF-β1 was decreased in each dose group of FAJ. Conclusion:FAJ has a protective effect on DMN-induced liver fibrosis in rats and was dose-dependent. Its mechanism of action may be related to anti-oxidation and inhibition of hepatic stellate cell activation.
Objective:To study the protective effect and mechanism of flavonoids from Ardisia japonica(Hornsted)BL.(FAJ)on dimethylnitrosamine(DMN)-induced hepatic fibrosis in rats,so as to provide scientific basis for clinical rational drug use and resource development in the future. Methods:Rats were randomly divided into normal group,model group,colchicine group(0.1 mg/kg),FAJ(600 mg/kg,400 mg/kg,200 mg/kg) groups. Except for normal group,each group was intraperitoneally injected with 0.5% DMN to establish hepatic fibrosis model in rats,and FAJ intervention was performed once a day until the end of the experiment. At the end of the experiment,blood and liver tissue samples were collected from the rats. Serum alanine aminotransferase(ALT),aspartate aminotransferase(AST),superoxide dismutase(SOD) and malondialdehyde(MDA),andhyaluronan were detected in each group of rats. The level of acid(HA)and laminin(LN)was detected by immunohistochemistry.The expression of transforming growth factor-β1(TGF-β1)in liver of rats in each group was examined. Histopathological changes of liver in rats of each group were observed by pathology. And protective effect and mechanism of FAJ on DMN-induced hepatic fibrosis in rats was investigated. Results :Compared with the model group,the serum levels of ALT,AST,MDA,HA,and LN in the FAJ(600 mg/kg,400 mg/kg) group were significantly lowed(P <0.01,P <0.05),and the SOD level was significantly increased(P <0.01),the serum levels of ALT,AST,MDA,HA,and LN in the FAJ(200 mg/kg) were significantly lowed(P <0.01,P <0.05),the level of SOD was increased,but there was no significant difference(P >0.05).Pathological sections showed that the degree of inflammatory cell infiltration and fibrosis in liver tissue was significantly improved after FAJ intervention. Immunohistochemical images showed that the expression of TGF-β1 was decreased in each dose group of FAJ. Conclusion:FAJ has a protective effect on DMN-induced liver fibrosis in rats and was dose-dependent. Its mechanism of action may be related to anti-oxidation and inhibition of hepatic stellate cell activation.
引文
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[9]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.
[10]高雅,韦日明,黄思茂,等.基于TGF-β_1/Notch信号通路研究杠板归对二甲基亚硝胺诱导大鼠肝纤维化的作用机制[J].中国医院药学杂志,2017,37(23):2318-2321.
[11]赵庆华,史海立,刘静生,等.强肝软坚丸对肝纤维化大鼠肝组织中MMP、TIMP表达的影响[J].中医学报,2017,32(2):260-264.
[12]朱跃科,段钟平,王宝恩,等.芪参益气滴丸对DMN大鼠肝纤维化及肝组织TIMP-1表达的影响[J].中国新药与临床杂志,2007,26(1):11-16.
[13]林远灿,骆海莺,金乾兴.垂盆草总黄酮影响肝星状细胞凋亡的作用机制研究[J].中国中药杂志,2015,40(16):3273-3277.
[14]黄思茂,曹后康,高雅,等.金花茶多糖对四氯化碳致小鼠急性肝损伤的保护作用及其机制的研究[J].中药药理与临床,2016,32(6):117-120.
[15]孙翠翠,康莲莲,王赛,等.藏药绿萝花抗氧化作用实验研究[J].中兽医医药杂志,2015,5(5):18-23.
[2]童家赟,梁之桃,赵中振,等.本草著作中紫金牛属药物基原考[J].中国中药杂志,2017,42(2):396-404.
[3]王福根,庄让笑,方红英,等.岩柏草总黄酮抗大鼠肝纤维化的实验研究[J].中国临床药理学与治疗学,2011,16(4):371-374.
[4]马官英,张庆月,钟瑞华,等.金银花总黄酮对氧化应激中肝星状细胞的保护作用[J].中国药房,2014,25(31):2895-2897.
[5]毕明,居靖,吴繁荣,等.野菊花总黄酮对肝纤维化大鼠治疗作用及机制探讨[J].中国药学杂志,2014,49(5):367-370.
[6]高雅,张可兰,黄思茂,等.狗肝菜多糖对二甲基亚硝胺诱导的肝纤维化大鼠TGF-β1/Smads信号通路的影响[J].中药药理与临床,2016,32(6):113-117.
[7]杨波,吴元凯,陈忠诚,等.ALT、AST、TBil及PT变化趋势与慢加急性乙型肝炎肝衰竭预后的关系[J].临床肝胆病杂志,2012,28(3):205-208.
[8]张可锋,黄思茂,陈毅飞,等.七味净肝灵对二甲基亚硝胺诱导肝纤维化大鼠的保护作用及其作用机制[J].中药材,2018(2):464-467.
[9]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.
[10]高雅,韦日明,黄思茂,等.基于TGF-β_1/Notch信号通路研究杠板归对二甲基亚硝胺诱导大鼠肝纤维化的作用机制[J].中国医院药学杂志,2017,37(23):2318-2321.
[11]赵庆华,史海立,刘静生,等.强肝软坚丸对肝纤维化大鼠肝组织中MMP、TIMP表达的影响[J].中医学报,2017,32(2):260-264.
[12]朱跃科,段钟平,王宝恩,等.芪参益气滴丸对DMN大鼠肝纤维化及肝组织TIMP-1表达的影响[J].中国新药与临床杂志,2007,26(1):11-16.
[13]林远灿,骆海莺,金乾兴.垂盆草总黄酮影响肝星状细胞凋亡的作用机制研究[J].中国中药杂志,2015,40(16):3273-3277.
[14]黄思茂,曹后康,高雅,等.金花茶多糖对四氯化碳致小鼠急性肝损伤的保护作用及其机制的研究[J].中药药理与临床,2016,32(6):117-120.
[15]孙翠翠,康莲莲,王赛,等.藏药绿萝花抗氧化作用实验研究[J].中兽医医药杂志,2015,5(5):18-23.