低分子肝素钠对小鼠实验性肝纤维化干预作用的初步研究

Interventional effect of low molecular weight heparin on mice with experimental hepatic fibrosis

目的:探讨低分子肝素钠(LMWH)对四氯化碳(CCl4)诱导的小鼠肝纤维化的干预作用。方法:将30只KM小鼠随机均分为空白对照组(腹腔注射橄榄油8周,n=10)、模型对照组(CCl4腹腔注射8周,n=10)和LMWH治疗组(CCl4腹腔注射+LMWH皮下注射8周,n=10)。8周后处死所有小鼠,收集血清及肝组织样本。检测血清丙氨酸转氨酶(ALT)、天冬氨酸转氨酶(AST)、总胆红素(TBIL)、直接胆红素(DBIL)和透明质酸(HA)水平;行HE染色、Masson三色染色和PSR染色观察肝组织病理形态及胶原纤维沉积情况;免疫组化检测肝组织α-平滑肌肌动蛋白(α-SMA)表达量;实时聚合酶链反应(RT-PCR)检测小鼠肝组织中纤维相关因子α-SMA和Ⅰ型胶原(COL-Ⅰ)mRNA表达水平。结果:与空白对照组比较,8周后模型对照组和LMWH治疗组小鼠血清ALT、AST、TBIL、DBIL和HA水平明显升高,肝组织胶原纤维沉积增多,肝组织α-SMA表达增多,α-SMA和COL-ⅠmRNA表达水平显著升高(均P<0.05)。与模型对照组比较,8周后LMWH治疗组小鼠血清ALT、AST、TBIL、DBIL和HA水平降低,肝组织胶原纤维沉积减少,肝组织α-SMA表达减少,肝组织α-SMA和COL-ⅠmRNA表达降低(均P<0.05)。结论:LMWH可抑制CCl4诱导的小鼠肝脏损害及纤维化,其机制可能与LMWH的抗凝作用有关。

AIM : To investigate the interventional effect of low molecular weight heparin on mice with hepatic fibrosis caused by carbon tetrachloride (CCl 4). METHODS : Thirty mice were randomly divided into three groups: normal control group (intraperitoneally injection of olive oil for 8 weeks, n =10),model group (intraperitoneally injection of CCl 4 for 8 weeks, n =10) and interventional group (intraperitoneally injection of CCl 4 and subcutaneous injection of low molecular weight heparin for 8 weeks, n =10).All animals were sacrificed at the end of week 8,and blood samples and liver tissue samples were obtained.The serum levels of alanine aminotransferase (ALT),aspartate aminotransferase (AST),total bilirubin (TBIL),direct bilirubin (DBIL) and hyaluronicacid (HA) were detected.Liver pathology morphology and collagen deposition were observed by HE staining,masson-trichrome staining and PCR staining.The expression of α-smooth muscle actin (α-SMA) in hepatic tissue was detected by immunohistochemistry.The expression of α-SMA and collagen type Ⅰ(COL-Ⅰ) mRNA in the hepatic tissue was quantified by real time PCR. RESULTS :Compared with control group,the serum level of ALT,AST,TBIL,DBIL and HA,the liver collagen deposition,the hepatic tissue expression of α-SMA and the hepatic tissue expression of α-SMA and COL-Ⅰ mRNA in both model group and interventional group were all significantly higher(all P < 0.05 ).Compared with model group,the serum level of ALT,AST,TBIL,DBIL and HA,the liver collagen deposition,the hepatic tissue expression of α-SMA and the hepatic tissue expression of α-SMA and COL-Ⅰ mRNA in interventional group were all significantly decreased (all P < 0.05 ). CONCLUSION : Low molecular weight heparin can attenuate the progression of mice hepatic damage and fibrosis caused by CCl 4.The mechanism may be related with the anticoagulation of low molecular weight heparin.