葛黄颗粒对酒精性肝病大鼠抗氧化应激及调节脂质代谢的作用机制

Mechanisms of antioxidative stress and lipid metabolism of Ge Huang Granules in rats of ALD model

目的:探讨葛黄颗粒对酒精性肝病(ALD)大鼠抗氧化应激及调节脂质代谢的作用机制。方法:108只SD大鼠随机分为6组:正常组、模型组、葛黄颗粒高、中、低剂量组、对照组(美他多辛),给予40%酒精(1.0 mL/100 g·d),同时隔日给予高脂饲料。4周后高、中、低剂量组分别给予不同剂量的葛黄颗粒(5.3、2.65、1.325 g/kg·d),对照组加用美他多辛(0.09 g/kg·d),持续8周。用RT-PCR法检测肝细胞过氧化物酶体增殖物激活受体-α的表达及肝细胞色素P4502E1的表达,用Western Blot测定肝细胞色素P4502E1的蛋白表达水平,用试剂盒测定肝组织丙二醛(MDA)的含量、超氧化物歧化酶(SOD)和谷胱甘肽过氧化物酶(GSH-PX)的活力。结果:与模型组比较,高剂量组和对照组大鼠肝组织中的MDA水平及P4502E1的基因表达和蛋白表达明显降低(P<0.05),而SOD、GSH-PX的活力以及PPAR-αmRNA的表达则显著增加(P<0.05),二组之间无明显差异(P>0.05)。结论 :葛黄颗粒能降低大鼠肝组织中P4502E1 mRNA及其蛋白的表达、MDA含量,增加PPAR-αmRNA的表达、SOD和GHS-PX的活力,表明其具有抗氧化应激及调节脂质代谢的作用,能有效地防治ALD。

Objective： To study the mechanisms of antioxidative stress and lipid metabolism of Ge Huang Granules （GHG） in rats of ALD model. Methods： 108 SD rat were randomly divided into 6 groups： normal group, model group, low,medium and high dosage GHG treatment groups, and control group （metadoxine）. Rats in ALD model groups were given 40% alcohol （1.0 mL/100g.d） by gavage and all mice were feed a high fat diet every other day. Four weeks later, rats in the the low, middle and high dosage groups were administrated with GHG at 5.3 g, 2.65 g, 1.325 g/kg-d respectively. Metadoxine was used for control group at 0.09 g/kg·d. Drug interventions were continued for 8 weeks. Real-time PCR was employed to analyze the RNA level of PPAR-a and P4502E1. For P4502E1, western blot was also used to determine the protein level. The levels of malondialdehyde （MDA）, superoxide dismutase （SOD）, and glutathione peroxidase （GSH-PX） in the liver tissue were measured. Results： Compared with the model group, high dosage of GHG and metadoxine significantly reduced the contents of MDA and the expression of P4502E1 at both mRNA and protein levels （P 〈 0.05）, and increased GSH levels, SOD, GSH-PX activities and PPAR-ot mRNA （P 〈 0.05） in liver. Conclusion： GHG can decrease levels of P4502E1 mRNA and protein and MDA level, and increase the PPAR-α mRNA, SOD and GHS-PX levels, suggesting that the therapeutic effect of GHG on ALD may be attributed to the altered oxidative stress and lipid metabolism.