清热利湿方对脂肪肝细胞磷脂肌醇-3激酶/肾上腺受体激酶表达的调节作用研究

Regulation of Qingre Lishi Formula on PI3K/ARK expression in fatty liver cells

目的观察清热利湿方对氧化损伤脂肪肝大鼠磷脂肌醇-3激酶(PI3K)/肾上腺受体激酶(ARK)通路调节作用。方法选取大鼠60只,分为正常对照组、高脂模型组、多烯磷脂酰胆碱组及清热利湿方高、中、低剂量组6组,采用高脂饲料复制大鼠脂肪肝实验动物模型,从第6周开始,清热利湿高、中、低剂量组分别给予0.3、0.1、0.05 g/(100 g·d)清热利湿方汤剂灌胃,多烯磷脂酰胆碱组给予25 mg/(100 g·d)灌胃,12周后取材。病理检查各组脂肪肝程度,ELISA检测血清高敏C反应蛋白(hs-CRP)含量及三磷酸腺苷酶(ATPase)活性,生化分析仪检测各实验组血脂及肝功能指标。western blot检测脂肪变肝细胞PI3K/ARK表达。结果高脂模型组hs-CRP明显升高,与清热高剂量组比较差异具有统计学意义(P=0.0331)。各实验组ATPase呈现不同程度降低,清热高剂量组血清ATPase水平明显提高,与高脂模型组比较差异具有统计学意义(P=0.0401);western blot实验结果可见,各实验组大鼠肝细胞PI3K/ARK表达存在明显差异,清热利湿方治疗后,PI3K/ARK水平明显降低。结论清热利湿方通过调节ATPase、hs-CRP、PI3K/ARK而降低肝细胞氧化应激炎症介质,改善肝细胞由于脂类代谢异常形成的过氧化损伤,促进肝细胞功能的恢复,具有一定的改善脂肪变性效果。

Objective To observe the regulative effect of Qingre Lishi Formula on PI3 K/ARK pathway of fatty liver rats with peroxidation damage. Methods 60 rats were randomly divided into 6 groups, including normal control group, high-fat model group, polyene phosphatidyl choline group, Qingre Lishi Formula low-dose, medium-dose, and high-dose group, and they were fed with high fat diet for establishing the model with fatty liver. From week 6, Qingre Lishi Formula groups were given the medicine respectively with 0.3/100 g/day,0.1 g/100 g/day, 0.05/100 g/day,polyene group with 24.6 mg/100 g/day,12 weeks out. Physiological check was given to observe the condition of fatty liver, ELISA was used to detect serum hs-CRP content and ATPase activity, biochemical analyzer to detect blood lipid and liver function indexes in each experimental group of fatty liver. Western-blot was used to detect the expression of PI3 K/ARK in fatty liver cells. Results The content of hs-CRP of high fat model group was increased significantly, compared with the Qingre Lishi Formula high dose group, the difference was statistically significant(P=0.0331). ATPase in the experimental groups showed decrease of different degree, serum level of ATPase of Qingre high dose group was significantly increased, and compared with hyperlipidemia model group, the difference was statistically significant(P=0.0401);Western blot experiment results indicated that PI3 K/ARK expression in liver cell of the experimental rats differed significantly, after Qingre Lishi Formula treatment, the expression level of PI3 K/ARK was significantly lowered. Conclusion Qingre Lishi Formula could improve peroxidation damage due to the formation of abnormal metabolism of lipid and promote the recovery of liver function through the regulation of ATPase, hs-CRP and PI3 K/ARK of inflammatory mediators in liver cells from oxidative stress and has certain effect of improving fatty degeneration.