【摘要】 目的观察不同剂量甲基阿魏酸(MFA)灌胃对大鼠酒精性肝病(ALD)的治疗作用,并探讨其机制。方法将60只SD大鼠随机分为低剂量组、中剂量组、高剂量组、模型组、正常组各12只,除正常组外,其余各组大鼠每天早上给予酒精灌胃1次,连续16周;从第13周开始,低剂量组、中剂量组、高剂量组每天下午分别灌胃给予5、10、20 mg/kg的MFA,模型组和正常组给予等体积的溶媒,连续4周。以16周后大鼠血清ALT、AST水平升高为构建ALD模型成功。16周后实验结束,处死大鼠,取血和肝脏组织,计算肝脏指数;取相同位置的肝脏组织,苏木精-伊红(HE)染色观察肝脏组织病变程度;生化分析法测定血清丙氨酸氨基转移酶(ALT)、天冬氨酸氨基转移酶(AST)和肝脏组织过氧化氢酶(CAT)、谷胱甘肽过氧化物酶(GSH-Px)、超氧化物歧化酶(SOD)、丙二醛(MDA)水平; Western blotting法检测肝脏组织Bax、Bcl-2、cleaved-Caspase-3、Caspase-3蛋白,计算Bax蛋白/Bcl-2蛋白、cleaved-Caspase-3蛋白/Caspase-3蛋白相对表达量比值; RT-PCR法检测肝脏组织Bax、Bcl-2 mRNA,计算Bax mRNA/Bcl-2 mRNA相对表达量比值。结果与模型组比较,低剂量组、中剂量组、高剂量组肝脏指数降低,肝脏组织病变改善,血清ALT和AST水平降低,肝脏组织CAT、GSH-Px、SOD水平降低,MDA水平升高,Bax蛋白/Bcl-2蛋白、Bax mRNA/Bcl-2 mRNA、cleaved-Caspase-3蛋白/Caspase-3蛋白相对表达量比值减少(P均<0. 05);低剂量组、中剂量组、高剂量组上述指标两两比较,P均<0. 05。结论 MFA对大鼠ALD有治疗作用,且高剂量效果更佳,其机制可能为减轻ALD发病关键环节氧化应激及细胞凋亡程度。更多还原

【Abstract】 Objective To observe the therapeutic effects of different doses of methyl ferulic acid( MFA) on alcoholic liver disease( ALD) in rats and to explore its mechanism. Methods Sixty SD rats were randomly divided into the lowdose group,medium-dose group,high-dose group,model group,and normal group,with 12 in each group. Except the normal group,rats in the other groups were given alcohol once a day for 16 weeks. From the 13 th week,the rats in the lowdose,medium-dose,and high-dose groups were given MFA at 5,10,and 20 mg/kg every afternoon,and the rats in the model group and normal group were given an equal volume of solvent for 4 weeks. After 16 weeks,the elevated rat serum ALT and AST levels were the criteria for the successful ALD models. After 16 weeks,the rats were sacrificed. The blood and liver tissues were collected and the liver index was calculated. The hepatic tissues at the same location were taken,and hematoxylin and eosin( HE) staining was used to observe the extent of liver lesions. Alanine aminotransferase( ALT),aspartate aminotransferase( AST) levels in serum and catalase( CAT),glutathione peroxidase( GSH-Px),superoxide dismutase( SOD) and malondialdehyde( MDA) levels in liver tissue were measured by biochemical analysis. Western blotting was used to detect the expression levels of Bax,Bcl-2,Cleaved-Caspase-3,and Caspase-3 protein in the liver tissues,and we calculated the ratio of Bax protein/Bcl-2 protein,cleaved-Caspase-3 protein/Caspase-3 protein. RT-PCR was used to detect the expression levels of Bax and Bcl-2 mRNA in the liver tissues,and the ratio of Bax mRNA/Bcl-2 mRNA was calculated. Results Compared with the model group,the liver index decreased,the liver tissue lesions were improved,the serum ALT and AST levels decreased,the liver tissue CAT,GSH-Px,SOD levels decreased,the MDA level increased,and the ratio of Bax protein/Bcl-2 protein,Bax mRNA/Bcl-2 mRNA,cleaved-Caspase-3 protein/Caspase-3 protein decreased in rats of the low-dose group,the medium-dose group,and the high-dose group( all P < 0. 05). Significant differences were found in the above indexes between the low-dose group,medium-dose group,and high-dose group( all P <0. 05). Conclusion MFA has a therapeutic effect on ALD in rats by reducing the degree of oxidative stress and apoptosis in the key part of ALD,and the higher the concentration,the better the effect.更多还原