清肝活血方及其拆方对酒精性肝病模型大鼠成纤维细胞生长因子-21影响

Effect of Qinggan-Huoxue recipe and its disassembled prescription on fibroblast growth factor-21 in rats with alcoholic liver disease

目的 观察清肝活血方及其拆方对酒精性肝病模型大鼠成纤维细胞生长因子-21（fibroblast growth factor21,FGF21）蛋白表达的影响,探讨其作用机制.方法 采用随机数字表法将大鼠分为空白组（n=10）、四氯化碳组（n=10）、模型组（n=11）、清肝活血方组（n=10）、清肝方组（n=9）、活血方组（n=10）.除空白组、四氯化碳组外,其余各组大鼠灌胃10 ml/kg酒精、玉米油、吡唑（4.5 ml：2 ml：25 mg）混合液,1次/d;腹腔注射0.3 ml/kg四氯化碳橄榄油溶液（四氯化碳：橄榄油=1：3）,2次/周,共给药12周,制备酒精性肝病模型.于第9周,清肝活血方组灌胃清肝活血方4.75 g/kg,清肝方组灌胃清肝方1.5 g/kg,活血方组灌胃活血方3.25 g/kg,1次/d,连续给药28 d.采用HE染色及Masson染色观察各组大鼠肝组织病理学改变;检测大鼠血清AST、ALT水平,采用免疫组化法、WB、荧光定量PCR检测肝组织FGF21蛋白和mRNA表达.结果 与模型组比较,清肝方组、活血方组、清肝活血方组肝重、肝体比明显降低（P〈0.01）,大鼠血清ALT[（41.95±26.78）U/L、（46.63±21.00）U/L、（37.57±27.85）U/L比（138.34±43.35）U/L]、AST[（102.74±23.55）U/L、（111.50±21.26）U/L、（83.72±33.81）U/L比（257.41±162.31）U/L]水平降低（P〈0.01或P〈0.05）;清肝方组、清肝活血方组FGF21蛋白[（1.19±0.07）、（1.24±0.12）比（0.92±0.04）]表达升高（P〈0.05）,清肝活血方组FGF21mRNA[（1.25±0.08）比（0.95±0.05）]表达增加（P〈0.05）.结论 清肝活血方可增加酒精性肝病模型大鼠肝组织FGF21蛋白及mRNA表达,保护肝组织.

Objective To investigate the mechanism of Qinggan-Huoxue recipe in the treatment of alcoholic liver disease by observing its function on alcoholic liver disease rat fibroblast growth factor -21 （FGF21）. Methods The rats were randomly divided into the blank group （n=10）, the carbon tetrachloride group （n=10）, the model group （n=11）, the Qinggan-Huoxue recipe group （n=10）, the Qinggan recipe group （n=9） and the Huoxue recipe group （n=10）. Except for the blank group and the carbon tetrachloride group, the other rats were given 10 ml/kg alcohol, corn oil and pyrazole mixture （4.5 ml： 2 ml： 25 mg）, once a day, 0.3 ml/kg carbon tetrachloride olive oil solution （carbon tetrachloride： olive oil =1： 3） intraperitoneally, and twice a week for 12 weeks to prepare the alcoholic liver disease model. In the ninth week, the Qinggan-Huoxue recipe group was treated with 4.75 g/kg of Qinggan-Huoxue recipe, the Qinggan recipe group was treated with 1.5 g/kg of Qinggan recipe, and the Huoxue recipe group was treated with 3.25 g/kg of Huoxue recipe, once a day, and 28 d for continuous administration. The pathological changes of liver tissue were observed by HE staining and Masson staining. The serum levels of AST and ALT were detected in rats. The changes of FGF21 protein and mRNA expression were detected by immunohistochemistry, Western blotting and fluorescence quantitative PCR. Results Compared with the model group, the liver weight and liver body ratio in the Qinggan recipe group, Qinggan-Huoxue recipe group and Huoxue recipe group were significantly lower （P〈0.01）. Compared with the model group, the serum ALT （41.95 ± 26.78 U/L, 46.63 ± 21.00 U/L, 37.57 ± 27.85 U/L vs. 138.34 ± 43.35 U/L）, AST （102.74 ± 23.55 U/L, 111.50 ± 21.26 U/L, 83.72 ± 37.57 U/L vs. 257.41 ± 162.31 U/L） in the Qinggan recipe group, Qinggan-Huoxue recipe group and Huoxue recipe group significantly decreased （P〈0.01 or P〈0.05）. Compared with the model group, the expression of FGF21 protein （1.19 ± 0.07, 1.24 ± 0.12 vs. 0.92 ± 0.04） in Qinggan recipe group and Qinggan-Huoxue recipe group significantly increased （P〈0.05）, and the expression of FGF21 mRNA （1.25 ± 0.08 vs. 0.95 ± 0.05） in Qinggan-Huoxue recipe group significantly increased （P〈0.05）. Conclusions FGF21 plays an important role in the alcoholic liver disease. During the process of liver injury, FGF21 increases continuously. Qinggan-Huoxue recipe can improve the content of FGF21 in the treatment of alcoholic liver disease.