目的 探讨扶肝化纤汤对肝纤维化模型大鼠MAPK信号通路的影响，研究其抗肝纤维化的作用机制。方法 120只SD大鼠，按照每组20只，随机分为对照组、模型组、秋水仙碱（2 mg/kg）组及扶肝化纤汤高、中、低剂量（67.08、33.54、16.77 g/kg）组。对照组不建模，其余各组在四氯化碳（CCl₄）诱导的大鼠肝纤维化模型的基础上，综合运用夹尾、强迫游泳、饥饱失常、ig大黄等方法，制备正虚毒蕴血瘀型肝纤维化大鼠模型。造模6周，进行肝病理检查，验证模型成功后，各给药组ig相应药物，对照组和模型组ig纯净水2 mL/d，连续给药3周。末次给药的次日，检测各组大鼠血清丙氨酸转氨酶（ALT）、天冬氨酸转氨酶（AST）、白蛋白（ALB）、羟脯氨酸（Hyp）、层黏连蛋白（LN）、Ⅲ型前胶原肽（PⅢNP）水平，肝组织行HE染色和Masson染色。采用Western blotting法检测肝组织MAPK信号通路中ERK1/2、JNK1/2、P38相关蛋白及其磷酸化蛋白表达情况，定量分析结果以p-ERK/ERK、p-JNK/JNK、p-p38/p38值表示。结果 模型组肝组织炎细胞浸润、纤维化程度重，各给药组肝损伤程度以及纤维化程度明显减轻。与对照组比较，模型组大鼠血清ALT、AST、Hyp、LN、PⅢNP明显升高，ALB明显下降（P<0.01），p-JNK、p-ERK、p-p38蛋白表达显著增高（P<0.01）。与模型组比较，扶肝化纤汤能降低大鼠血清ALT、AST、Hyp、LN、PⅢNP水平，升高ALB水平，下调p38、ERK1/2、JNK1/2的磷酸化蛋白表达，且呈现出剂量依赖性，以高剂量组效果最好。结论 正虚毒蕴血瘀病症结合肝纤维化模型大鼠建模成功。扶肝化纤汤具有保护肝细胞，减轻肝损伤，抑制肝纤维化的作用。扶肝化纤汤能够调控p-JNK、p-ERK、p-p38蛋白表达，抑制MAPK信号通路的活化，可能是其抗肝纤维化的作用机制之一。

Objective To research the effect of Fugan Huaxian Decoction on MAPK signaling pathway in rats with hepatic fibrosis (HF) and explore the mechanism of anti-HF. Methods A total of 120 SD rats were randomly divided into control group, model group, colchicine group and Fugan Huaxian Decoction group (high, medium and low dose groups), with 20 rats in each group. The rats in the normal group were SD rats, and the rats model in the remaining groups were established into HF rats with syndrome of qi deficiency, poison weakness and blood stasis on the basis of the HF model induced by carbon tetrachloride (CCl₄). Moreover, the rats were also received tail clamping, forced swimming, abnormal of starvation and full as well as rhubarb gavage. Liver pathology was performed on all rats after six weeks of modeling. After the validation model was successful, each group was given different doses of gavage, colchicine group (2 mg/kg), high, medium and low dose groups of Fugan Huaxian Decoction were intra-gastrically administered (67.08, 33.54, and 16.77 g/kg), normal group and model group were given pure water 2 mL/d for three weeks continuously. On the second day of last gavage, the serum levels of ALT, AST, ALB, LN, HyP and PⅢNP were detected in each group. HE staining and Masson staining were performed on liver tissues. The expression of ERK1/2, JNK1/2 and P38-related protein and their phosphorylated protein in liver MAPK signaling pathway were detected by Western blotting. The results of quantitative analysis were expressed as p-ERK/ERK, p-JNK/JNK, p-p38/p38 ratio. Results In model group, hepatic tissue cells were severely infiltrated and fibrotic, while, the degree of liver injury and fibrosis were significantly reduced in other groups. Compared with normal group, the serum levels of ALT, AST, Hyp, LN and PⅢNP in model group were increased significantly, ALB was decreased significantly (P < 0.01), and the protein expression of p-JNK, p-ERK and p-p38 was increased significantly (P < 0.01). Compared with model group, Fugan Huaxian Decoction decreased serum ALT, AST, Hyp, LN and PⅢNP levels of rats, increased ALB content and down-regulated the expression of phosphorylated protein of p38, ERK1/2 and JNK1/2, and it showed in a dose-dependent manner, the high dose group worked the best. Conclusion The model of qi deficiency, poison weakness and blood stasis combined with HF model rats was successfully established. Fugan Huaxian Decoction not only protects hepatocytes, alleviates liver injury and inhibits HF, but also regulates the protein expressions of p-JNK, p-ERK, p-p38, as well as inhibits MAPK signaling pathway activation, which may be one of the mechanisms of its anti-HF function.

R285.5

国家自然科学基金资助项目（81660790）；贵州省科学技术基金项目资助课题（黔科合LH字[2014]7318号）；院内博士启动基金课题