大柴胡汤调控TGF-β/Smad信号通路对DBTC联合乙醇诱发小鼠胰腺纤维化的防治作用  

Effect of DaChaiHu Decoction on pancreatic fibrosis induced by DBTC combined with alcohol and the mechanism of TGF-beta/Smad signaling pathway

作　　者：许小凡[1] 姜婷婷[2] 刘芳[3] 张晓芹[3] 史迎莉 陈瑜[3] 李涛[3] 张红 

机构地区：[1]陕西中医药大学医学科研实验中心,西安712046 [2]榆林市第一医院病理科,榆林719000 [3]陕西中医药大学基础医学院,西安712046

出　　处：《中国应用生理学杂志》2016年第5期444-448,482,共6页Chinese Journal of Applied Physiology

基　　金：国家自然科学基金(81102725);陕西省教育厅自然科学重点科研计划(15JS027)

摘　　要：目的:探讨大柴胡汤对二丁基二氯化锡(DBTC)联合乙醇所致小鼠慢性胰腺炎胰腺纤维化中TGF-β/Smad信号通路的影响及大柴胡汤防治胰腺纤维化的作用机制。方法:昆明小鼠96只,随机分成空白组(Con组)、慢性胰腺炎组(CP组)、大柴胡汤治疗组(DCHD组)(n=32)。一次性尾静脉注射DBTC(8 mg/kg)联合10%乙醇饲喂代替正常饮水复制小鼠CP模型。注射DBTC 3 d后小鼠又随机分为CP组和DCHD组,DCHD组予大柴胡汤(1 g/ml,6 g/kg·d)灌胃,并伴随10%乙醇饲喂代替正常饮水。各组在1、2、4、8周分批处死小鼠(n=8),观察胰腺组织的形态学变化及血清淀粉酶、透明质酸的变化;检测胰腺组织MMP-1/TIMP-1 mRNA的表达以及胰腺组织Ⅰ型胶原、TGF-βRⅠ、p-Smad 2/3、Smad 7蛋白的表达。结果:与空白组相比,CP组2周、4周血清淀粉酶及透明质酸处于较高水平,8周时淀粉酶明显降低,而透明质酸水平进一步升高(P<0.05);DCHD组血清淀粉酶及透明质酸含量明显低于CP组(P<0.01)。与空白组相比,CP组胰腺组织COLA1、TGF-βRⅠ、p-Smad 2/3表达持续升高,Smad7蛋白表达明显减少;DCHD治疗组可降低COLA1的表达水平有效抑制CP小鼠胰腺TGF-βRⅠ、p-Smad 2/3表达,使Smad7蛋白表达升高。与空白组相比,CP 2周、4周胰腺MMP-1 mRNA表达持续降低,TIMP-1 mRNA表达随造模时间延长明显升高(P<0.01);DCHD组各时间点胰腺MMP-1表达明显增多,TIMP-1表达减少(P<0.05)。结论:大柴胡汤通过抑制TGF-β/Smad信号通路活化,调节MMP-1/TIMP-1的平衡,发挥防治慢性胰腺炎胰腺纤维化的作用。Objective: To investigate the effects of transforming growth factor-β( TGF-β) and Smad in chronic pancreatitis( CP)induced by dibutyltin dichloride( DBTC) combined with ethanol,and the effect for prevention and treatment of pancreatic fibrosis by Da Chai Hu Decoction( tradeiional Chinese medicine). Methods: Ninety-six healthy male Kunming mice were randomly divided into control group( Con group),chronic pancreatitis( CP group) group,Dachai Hu decoction treatment group( DCHD group)( n = 32).Then the mice were treated with DBTC 8 mg / kg by tail vein injection and fed with 10% ethanol replacing the normal drinking water to replicate mouse CP model. After three days of modeling the mice were randomly divided into CP group and DCHD group. The mice in DCHD group were administered intragastrically with Da Chai Hu Decoction( 1 g / ml,6 g / kg·d) at the following 8 weeks. After modeling for 1 week,2 weeks,4 weeks and 8 weeks,the mice were anesthetized and sacrificed( n = 8). The morphology and the degree of fibrosis changes of pancreatic tissue were detected by HE staining. The protein expressions of type I collagen and TGF beta R I,pSmad 2 /3 and Smad 7 in pancreatic tissue were detected by Western blot assay. The expressions of MMP-1 / TIMP-1 mRNA in pancreatic tissue were detected by RT-PCR. Results: Compared with control group,DBTC combined with ethanol induced CP with increased activity of serum amylase and hyaluronic acid level. While in DCHD group,the activity of amylase and hyaluronic acid level were decreased significantly. Compared with control group,the protein expressions of COLA1,TGF beta R I,p-Smad 2 /3 in CP Group were elevated,but the expression of Smad 7 was significantly reduced; In DCHD treatment group,the expression of TGF beta R I,p-Smad2 /3 and COLA1 were reduced,and the protein expression of Smad 7 was increased. In 2w and 4w,the level of MMP-1mRNA continued to decrease,while TIMP-1mRNA expression was increased significantly( P < 0. 01). At each time point of DCHD group,the expression of MMP-1 w

关 键 词：胰腺纤维化 大柴胡汤 TGF-Β/SMAD 细胞外基质 DBTC联合乙醇 

分 类 号：R285.5[医药卫生—中药学]