夏枯草总三萜调控ERK、TGF-β1/Smad通路对肝纤维化大鼠的保护作用研究  

Regulatory effects of total triterpenoid of Prunella vulgarisL. on activities of ERK and TGF-β1 / Smad signaling pathway in protecting hepatic fibrosis in rats

作　　者：章圣朋[1,2] 何勇[1,3] 徐涛[1,3] 黄成[1,3] 谢加力 邓子煜[5] 李俊[1,3] 

机构地区：[1]安徽医科大学药学院,安徽合肥230032 [2]皖南医学院药学院,安徽芜湖241002 [3]安徽省创新药物产业共性研究院,安徽合肥230032 [4]芜湖市第二人民医院药剂科,安徽芜湖241001 [5]安徽医科大学第二附属医院药剂科,安徽合肥230601

出　　处：《中国药理学通报》2015年第2期261-266,共6页Chinese Pharmacological Bulletin

基　　金：国家自然科学基金资助项目(No 81273526);安徽省科技厅项目(No KJ2012A156)

摘　　要：目的研究夏枯草总三萜(TTP)对四氯化碳(CCl4)致肝纤维化大鼠的保护作用及其分子机制。方法 SD大鼠随机分为正常组、模型组、TTP(25、50、100 mg·kg-1)组和阳性对照组(秋水仙碱0.1 mg·kg-1),除正常组外,其余各组分别于大鼠背部皮下注射CCl40.1 ml·(100 g)-1,每周2次,连续12周,自造模第5周起开始给药,各给药组分别给予相应的药物,正常组、模型组给予等体积的溶媒,每天1次。实验结束后,比色法测定血清中丙氨酸氨基转氨酶(ALT)、天冬氨酸氨基转移酶(AST)含量;放免法测定透明质酸(HA)、Ⅲ型前胶原(PCⅢ)、Ⅳ型胶原(CⅣ)含量;同时取固定部位肝组织,苏木精伊红染色(HE)、Masson染色观察肝纤维化程度;制备100 g·L-1肝匀浆,进行丙二醛(MDA)、超(过)氧化物歧化酶(SOD)、谷胱甘肽过氧化物酶(GSH-Px)、羟脯氨酸(Hyp)含量测定;RT-PCR法检测α-SMA、Procollagen I、Smad2、Smad3和Smad7 mRNA表达;Western blot法检测pERK蛋白表达。结果与模型组相比,TTP(25、50、100 mg·kg-1)给药组不仅能降低肝纤维化大鼠ALT、AST、HA、CⅣ、PCⅢ、Hyp水平,改善肝脏病变程度,降低MDA水平,增强SOD以及GSH-Px活性,还可抑制肝组织中α-SMA、Procollagen I、Smad2、Smad3及p-ERK表达,升高Smad7表达。结论夏枯草总三萜对CCl4诱导的肝纤维化大鼠具有较好的保护作用,其机制可能与下调p-ERK表达,调控TGF-β1/Smad信号通路有关。Aim To investigate the protective effectsof total triterpenoid from Prunella vulgaris L.( TTP) onCCl4-induced hepatic fibrosis in rats and its mechanism. Methods Rat liver fibrosis was induced by50% CCl4 twice a week for 12 weeks. From the 5th week,all the therapeutic groups were treated with the TTP( 25,50,100 mg·kg- 1) and the colchicine( 0. 1mg· kg- 1) respectively once a day for 8 weeks. At the end of the twelfth week,the levels of ALT,AST,HA,PC Ⅲ,C Ⅳ,MDA,SOD,GSH-Px,Hyp were measured. HE and Masson staining were used to evalulate the degree of hepatic fibrosis. The mRNA expression of α-SMA,procollagen I,Smad2,Smad3,Smad7 in liver was detected by RT-PCR,and the p-ERK protein expression was evaluated by Western blot. Results Compared with the model group,TTP( 25,50,100mg·kg- 1) not only reduced serum content of ALT,AST,HA,PCⅢ,CⅣ and Hyp,MDA in liver tissue,improved the morphologic changes of hepatic fibrosis,but also increased SOD and GSH-Px activity. Moreover,it decreased the α-SMA,procollagen I,Smad2,Smad3 mRNA expression and increased Smad7 mRNA expression in liver tissues obviously. Furthermore,TTP reduced the protein expression of p-ERK. Conclusions TTP can protect rats from CCl4-induced liver fibrosis. The mechanism of this process may involve inhibiting the expression of p-ERK and interference with TGF-β1 / Smad signal transduction pathway.

关 键 词：夏枯草总三萜 肝纤维化 四氯化碳 脂质过氧化 TGF-Β1/SMAD信号通路 大鼠 

分 类 号：R-332[医药卫生] R284.1