基于MAPK信号通路研究4-羟基苯并恶唑-2-酮抗肝纤维化的作用机制

目的： 探讨4-羟基苯并恶唑-2-酮（HBOA）对四氯化碳（CCl₄）诱导的大鼠肝纤维化的保护作用。方法： 采用CCl₄橄榄油溶液灌胃建立大鼠肝纤维化模型。第8周确定建模成功后，将雄性SD大鼠随机分成正常对照组，模型对照组，秋水仙碱阳性药组，HBOA高、中、低药物组。从第9周起，相应药物干预4周后，测定血清中总蛋白（TP），白蛋白（Alb）的含量以及肝组织中羟脯氨酸（Hyp）的含量；免疫组织法检测肝组织中α-平滑肌肌动蛋白（α-SMA）蛋白含量的表达；Westerns blot法检测肝组织中MAPK，P-MAPK，P38的蛋白含量的表达。结果： 与模型组相比，HBOA可降低肝纤维化大鼠血清TP、Alb水平（P<0.01）以及肝组织中Hyp水平（P<0.01），HBOA高中低剂量组显著降低大鼠肝组织中的α-SMA蛋白的表达（P<0.01），并显著下调MAPK，P-MAPK，P38的蛋白表达水平（P<0.01）。结论： HBOA对CCl₄诱导的肝纤维化大鼠有一定的改善作用，通过抑制MAPK的信号通路，显著减轻了CCl₄诱导的纤维化。

The effect and mechanism of 4-hydroxy-2(3H)-benzoxazolone on MAPK signaling pathway of anti-hepatic fibrosis in rats

OBJECTIVE To explore the potential mechanism by which 4-hydroxy-2(3H)-benzoxazolone (HBOA) protects against hepatic fibrosis in rats induced by CCl₄.METHODS A hepatic fibrosis model of rats was established by orally administering 50% CCl₄/olive oil for 12 weeks. Then, at the 8th week, the successful model rats were randomly divided into a normal control group, a hepatic fibrosis model group, a colchicine group and three dose HBOA treatment groups. From the 9th week, animals were treated with the drugs daily for 4 weeks. The changes of the serum indicators including total protein (TP), albumin (Alb) and liver tissue indicators hydroxyproline (Hyp) were determined. Moreover, the protein of α-SMA in liver was examined by immunohistochemistry. The expressions of MAPK, P-MAPK and P38 protein were detected with Western blot.RESULTS Compared with the model group, the levels of TP and Alb in the rat serum and Hyp in liver tissues of HBOA groups were significantly decreased (P<0.01). The protein expressions of α-SMA in liver tissues of HBOA groups (100, 75, 50 mg·kg^-1) were decreased (P<0.01), and the expressions of MAPK, P-MAPK and P38 protein were also down regulated by HBOA (P<0.01).CONCLUSION HBOA have beneficial effects against liver fibrosis in rats induced by CCl₄.The present study results indicate that HBOA significantly alleviates CCl₄-induced liver fibrosis through suppressing the MAPK signaling pathway.