穿山龙水提物调控Keap1/Nrf2通路抗CCl_4诱导小鼠急性肝损伤作用研究

Study on the Effects of Aqueous Extract from Dioscorea Nipponica Makino Against Carbon Tetrachloride-Induced Mice Acute Liver Injury via Regulating Keap1/Nrf2 Signal Pathway

目的:评价穿山龙水提物(AEDN)通过对Keap1/Nrf2信号通路的调控实现对CCl_4诱导小鼠急性肝损伤的保护作用。方法:穿山龙药材经水提取浓缩蒸干即为AEDN,采用紫外可见分光光度法测定提取物中多糖含量。雄性昆明小鼠经AEDN连续灌胃给药7 d后,腹腔注射0. 35%CCl_4橄榄油溶液诱导急性肝损伤,评价AEDN对CCl_4诱导急性肝损伤氧化应激相关指标的影响,并检测AEDN对Keap1/Nrf2信号通路的调控作用。结果:AEDN的提取率为9. 64%,其中多糖含量为(53. 03±0. 70)%。在CCl_4所致小鼠急性肝损伤中,AEDN能显著降低MDA的表达水平,使小鼠肝组织中的SOD、GSH和GSH-Px含量增多。此外,AEDN可显著上调SOD1、SOD2、Nrf2、GST、NQO1和HO-1的蛋白表达,降低Keap1的表达,显著下调iNOS mRNA的表达水平。结论:穿山龙水提物可通过调控Keap1/Nrf2氧化应激信号通路实现对CCl_4诱导的小鼠急性肝损伤的保护作用。

Objective To evaluate the protective effect of the aqueous extracted from Dioscorea nipponica Makino （AEDN） on carbon tetrachloride （CCl 4）-induced acute liver injury in mice by regulating Keap1/Nrf2 signal pathway. Methods： Dioscorea nipponica Makino was extracted and concentrated by water and dried to be powder,which was AEDN.The content of polysaccharide in AEDN was determined by ultraviolet visible （UV） spectrophotometery.The male Kunming mice were adiminstrated with AEDN for 7 days,and then acute liver injury was induced by intraperitoneal injection of 0.3% CCl 4 olive oil solution.The effect of AEDN on oxidative stress indexes in CCl 4 induced liver injury mice and the regulation effect of AEDN on Keap1/Nrf2 signal pathway were evaluated. Results： The extraction rate of AEDN was 9.64%,and the polysaccharide content was （53.03±0.70）%.In CCl 4-induced mice actue liver injury,AEDN significantly decreased the level of MDA and increased the level of SOD,GSH and GSH-Px in liver tissue of mice.The mechanism of action study found that the protein levels of T SOD1,SOD2,Nrf2,GST,NQO1 and HO-1 were all markedly up-regulated,and the level of Keap1 was down-regulated.The mRNA level of iNOS were also suppressed by AEDN treatment. Conclusion： AEDN has protective effect on CCl 4-induced acute liver injury in mice via adjusting Keap1/Nrf2 signaling pathway.