丹酚酸A通过调节NF-κB/IκBα信号通路抑制肝纤维化

Anti-fibrotic effect of salvianolic acid A on hepatic fibrosis in rats by targeting NF-κB/IκBα signaling pathway

目的探索丹酚酸A抗四氯化碳(CCl_4)诱导的大鼠肝纤维化的防治及作用机制。方法将48只SD大鼠随机分为空白对照组、CCl_4模型组、丹酚酸A低剂量给药组和丹酚酸A高剂量给药组。除空白对照组外,其他3组每周2次给予50%CCl_4/花生油溶液(1 m L·kg-1)灌胃诱导建立肝纤维化模型,丹酚酸A低剂量和高剂量给药组大鼠同时每日1次给予腹腔注射丹酚酸A(分别为5 mg·kg-1与15 mg·kg-1)。6周后处死大鼠,HE和Masson染色法进行肝组织病理学观察;全自动生化分析仪检测血清肝功能指标(谷丙转氨酶及谷草转氨酶);放射免疫法测定血清肝纤维化指标(透明质酸、Ⅳ型胶原、层黏蛋白和Ⅲ型前胶原肽);Western blot检测肝脏组织中NF-κB和IκBα蛋白的表达;Q-PCR检测炎症因子以及肝星状细胞激活因子基因水平的表达。结果与CCl_4模型组相比,丹酚酸A给药组能够显著改善大鼠肝功能,降低大鼠肝纤维化程度(P<0.05)。与此同时,丹酚酸A给药组能够提高肝细胞浆中NF-κB、IκBα蛋白的表达,并降低细胞核NF-κB蛋白的表达,且能够抑制炎症因子与肝星状细胞激活因子基因水平的表达。结论丹酚酸A可通过调节NF-κB/IκBα信号通路发挥抗肝纤维化作用。

Objective To investigate the anti-fibrotic effect of salvianolic acid A on hepatic fibrosis in rats and the pharmacological mechanism of anti-fibrosis. Methods Forty-eight male Sprague-Dawley （SD） rats were randomly divided into 4 groups： a blank control group, a CC14model group, a salvianolic acid A low dose group （5 mg kg -1）, and a salvianolic acid A high dose group （15 mg kg -1）. Except for the blank control group, the rats in the CC14 model group and the 2 salvianolic acid A treatment groups were intragastrically administered solution containing 50% CC14 and peanut oil with the volume proportion of 1 ： 1 twice a week for 6 weeks to establish liver fibrosis model. Besides establishing the hepatic fibrosis model, the rats in the 2 treatment groups were given salvianolic acid A via intraperitoneal injection once daily according to the daily body weight after the first administration of CC14, respectively. HE and Masson＇s trichrome staining were done after six weeks. At the same time, liver function markers （alanine transaminase and aspartate aminotransferase） were detected by full- automatic biochemistry analyzer, and hepatic fibrosis markers （hyaluronic acid, collagen Type IV, laminin and procollagen HI peptide ） by radioimmunoassay. In addition, Western blot and Q-PCR were used to determine the expression levels of related protein and genes correspondingly. Results Compared with the CC14 model group, theliver function and degree of liver fibrosis were both improved significantly （P 〈 0.05） in the salvianolic acid A groups. Salvianolic acid A increased the expression levels of NF-κB and IκBα in the cytoplasm, and decreased the expression level of NF-κB in the cell nucleus. Meanwhile, salvianolic acid A inhibited the gene expression levels of inflammatory factors and makers related to HSCs. Conclusion Salvianolic acid A can inhibit hepatic fibrosis by targeting NF-κB/IκBα signaling pathway.