葛根素对四氯化碳所诱导肝纤维化大鼠的干预作用及对TLR-4、NF-κB、AP-1的影响

目的：研究葛根素对四氯化碳所诱导肝纤维化大鼠的干预作用及其对TLR-4、NF-κB、AP-1的影响。方法：将56只SD大鼠腹腔注射四氯化碳花生油溶液（1：1）1 mL·kg^-1，每周2次，连续注射8周，建立肝纤维化大鼠模型。将经病理学检查确认成肝纤维化模型的大鼠随机分为5组：模型组、阳性药水飞蓟素40 mg·kg^-1组、葛根素20，40，80 mg·kg^-1组，每组10只。每日灌胃给药一次，连续给药60 d。同时另设置正常组大鼠10只。Masson染色法观察各组肝组织病理学变化和纤维化程度。末次给药12 h后，检测大鼠血清谷草转氨酶（AST）、丙氨酸转氨酶（ALT）、白介素-6（IL-6）的水平；Elisa法检测肝组织中肿瘤坏死因子-α（TNF-α）、白介素-1（IL-1）的含量；蛋白免疫印迹法检测肝组织中TLR-4、NF-κB、AP-1的蛋白表达。结果：与模型相比，葛根素可降低肝纤维化大鼠血清AST、ALT、IL-6的水平以及TNF-α、IL-6的含量（P<0.05），并明显下调TLR-4、NF-κB、AP-1的蛋白表达水平（P<0.05）。结论：葛根素具有一定的抗肝纤维化作用，其作用机制可能与调节肝组织中TLR-4、NF-κB、AP-1的蛋白水平有关。

Effect of puerarin on expression levels of TLR-4, NF-κB and AP-1 in carbon tetrachloride induced liver fibrosis rats

OBJECTIVE To investigate the effect of puerarin on expression levels of TLR-4, NF-κB and AP-1 in carbon tetrachloride induced liver fibrosis rats.METHODS Liver fibrosis was induced in fifty six SD rats by intraperitoneally injecting carbon tetrachloride (1 mL·kg^-1) solved in peanut oil (1:1) twice a week, for 8 weeks continuously. The rats with liver fibrosis confirmed by pathological examination were divided into following 5 groups with 10 animal per group:model control group, 40 mg·kg^-1 silymarin group as positive control, 20, 40, 80 mg·kg^-1 puerarin groups. The rats were dosed once a day for 60 days. Other 10 healthy rats were assigned as normal control. The pathological and fibrosis changes were observed through Masson staining. Twelve hours after last dosing, the serum levels of aspartate aminotransferase (AST), alanine aminotransferase (ALT) and interleukin-6 (IL-6) were detected. The ELISA method was used to determine the contents of tumor necrosis factor-α (TNF-α) and interleukin-1 (IL-1). Western blot method was utilized to determine the expressions of TLR-4, NF-κB and AP-1 proteins in liver tissues.RESULTS Compared with model control, serum levels of AST, ALT, IL-6, TNF-α and IL-6 were significantly decreased by puerarin (P<0.05), and the expressions of TLR-4, NF-κB and AP-1 proteins were also down regulated by puerarin (P<0.05).CONCLUSION The antihepatic fibrosis mechanism of puerarin may be associated with the regulation of TLR-4, NF-κB and AP-1 proteins in liver tissues.