益气化瘀化痰养阴剂对肝纤维化大鼠FSP1、PAR-2表达的影响

Effects of Yiqi-huayu-huatan-yangyin preparation on the expression of fibroblast-specific protein 1 and protease activated receptor-2 in rats with liver fibrosis

摘要:

目的：观察益气化瘀化痰养阴剂对四氯化碳（CCl4）诱导的肝纤维化大鼠成纤维细胞特异性蛋白1（fibroblast-specific protein 1，FSP1）和蛋白酶激活受体-2（protease activated receptor，PAR-2）表达的影响。方法：将大鼠随机分为正常组、模型组、秋水仙碱组、益气化瘀化痰养阴剂低、中、高剂量组（分别简称为中药低、中、高剂量组）。正常对照组、模型组灌服生理盐水；秋水仙碱组灌服秋水仙碱0.1 mg/（kg?d）；中药低、中、高剂量组分别灌服益气化瘀化痰养阴剂[相当于生药30、60、120 g/（kg?d）]。连续给药12周后，观察各组动物的肝组织病理学变化，并检测血清丙氨酸氨基转移酶（alanine aminotransferase，ALT）、天门冬氨酸转移酶（aspartate aminotransferase，AST）、透明质酸（hyaluronan，HA）及Ⅳ型胶原（collagen Ⅳ，CⅣ）含量，用实时荧光定量PCR（qRT-PCR）和免疫印迹法（Western blot）检测各组大鼠肝组织FSP1和PAR-2的基因和蛋白表达。结果：（1）模型组大鼠与正常组大鼠相比，前者肝组织纤维化程度明显，血清中ALT、AST、HA、CⅣ含量明显增高（P<0.05），肝组织中FSP1和PAR-2的mRNA和蛋白表达亦明显增高（P<0.05）。（2）各治疗组与模型组相比，前者肝组织炎性细胞浸润少，肝纤维化程度轻，血清中ALT、AST、HA、CⅣ含量仅轻度升高（P<0.05），肝组织中FSP1和PAR-2的mRNA和蛋白表达亦轻度升高（P<0.05）。结论：益气化瘀化痰养阴剂能保护肝脏，延缓大鼠肝组织纤维化进程，其机制可能与调控大鼠肝组织FSP1和PAR-2的基因和蛋白表达有关。

Abstract:

Objective：To observe the effects of Yiqi-huayu-huatan-yangyin preparation on the liver histopathology and expression of fibroblast-specific protein 1（FSP1） and protease activated receptor -2（PAR-2） in liver fibrosis rats induced by CCl4. Methods：The SD rats were randomly divided into：normal group，model group，colchicines group and low/moderate /high dosages of Yiqi-huayu-hu－atan-yangyin preparation group. Rats in normal and model group were fed with normal saline，rats in colchicine group were fed with 0.1 mg/（kg·d） colchicine and rats in low/moderate/high dosages of Yiqi-huayu-huatan-yangyin preparation group were fed with Yiqi-huayu-huatan-yangyin[30，60，120 g/（kg·d）] correspondingly.Pathological changes of the liver tissues were observed by light microscopy. Activities of alanine aminotransferase（ALT），aspartate aminotransferase（AST） in liver tissues were evaluated by spec－trophotometry. Levels of hyaluronic acid（HA），typeⅣ collagen（CⅣ） in serum were determined using ELISA kit. Expression of FSP1 and PAR-2 mRNA and protein in the liver tissues was detected by qRT-PCR and Western blot. Results：（1）Levels of ALT，AST，HA and CⅣ in serum and fibrosis changes were increased significantly in model group compared with those in normal group（P<0.05） and the expression of FSP1 and PAR-2 mRNA and pro－tein of liver tissues were increased obviously（P<0.05）. （2）In－flammatory cell infiltration and liver fibrosis were lighter in treatment groups than in model group. Levels of ALT，AST，HA and CⅣ in serum and expression of FSP1 and PAR-2 mRNA and protein in liver tissues were slightly elevated in treatment groups（P<0.05）. Conclusion：Yiqi-huayu-huatan-yangyin preparation may protect the liver and delay the development of liver fibrosis by regulating the mRNA and protein expression of FSP1 and PAR-2.