基于网络药理学探讨麦角硫因抗D-半乳糖诱导氧化损伤的作用机制

Mechanistic investigation of ergothioneine against D-galactose-induced oxidative damage based on network pharmacology

中文关键词:  麦角硫因  抗氧化  D-gal  网络药理学  作用机制

英文关键词:ergothioneine  antioxidant  D-galactose  network pharmacology  mechanism

中文摘要:

      目的 评估麦角硫因对D-半乳糖(D-galactose, D-gal)诱导的大鼠氧化损伤的保护作用, 并探讨其潜在作用机制。方法 采用腹腔注射D-gal (200 mg/kg)连续6周建立大鼠氧化损伤模型。SD雄性大鼠随机分为5组: 空白组腹腔注射生理盐水, 模型组及麦角硫因低(EGT-L, 1.4 mg/kg)、中(EGT-M, 2.8 mg/kg)、高剂量组(EGT-H, 5.6 mg/kg)腹腔注射D-gal。自第3~6周, 各给药组每日灌胃麦角硫因, 空白组与模型组灌胃等体积生理盐水, 连续4周。采用比色法检测大鼠血液和肝脏中脂质氧化产物标志物丙二醛(malondialdehyde, MDA)、蛋白质氧化产物标志物蛋白质羰基、抗氧化物质还原型谷胱甘肽(glutathione, GSH)和抗氧化酶标志物过氧化氢酶(catalase, CAT)、超氧化物歧化酶(superoxide dismutase, SOD)的水平。采用网络药理学初步研究麦角硫因抗氧化损伤的作用机制, 进一步利用蛋白质免疫印迹法验证麦角硫因对大鼠Acetyl-FOXO1、FOXO1和SIRT1蛋白表达水平的影响。结果 与模型组相比, 各给药组可不同程度地降低大鼠血液和肝脏的MDA和蛋白质羰基水平, 并增加SOD、CAT活性和GSH水平。Western Blotting实验结果显示, 给药后可降低Acetyl-FOXO1/FOXO1的蛋白表达量, 升高SIRT1的蛋白表达量。结论 麦角硫因给药对D-gal诱导的大鼠氧化损伤有显著的保护作用, 其潜在的作用机制可能主要与FoxO信号通路有关。

英文摘要:

      Objective To evaluate the protective effects of ergothioneine against D-galactose (D-gal)-induced oxidative damage in rats and elucidate the underlying molecular mechanisms. Methods Oxidative damage in rats was induced by intraperitoneal injection of D-gal (200 mg/kg) once daily for 6 consecutive weeks. Male SD rats were randomly assigned to 5 groups: The control group, which received intraperitoneal saline injection, and the model group as well as the low-dose (EGT-L, 1.4 mg/kg), mid-dose (EGT-M, 2.8 mg/kg) and high-dose (EGT-H, 5.6 mg/kg) ergothioneine groups, which were administered intraperitoneal D-gal. Starting from week 3 and continuing through week 6, each treatment group received daily intragastric administration of ergothioneine, while the control and model groups were given an equal volume of saline, for a total duration of 4 weeks. The levels of oxidative stress markers, including malondialdehyde (MDA) as a marker of lipid peroxidation and protein carbonyls as markers of protein oxidation, antioxidants such as reduced glutathione (GSH), and antioxidant enzymes including catalase (CAT) and superoxide dismutase (SOD) were measured in the serum and liver of rats using colorimetric assays. A preliminary network pharmacology analysis was performed to investigate the potential mechanisms underlying the antioxidant effects of ergothioneine, and further validation was carried out using western blotting to assess the impact of ergothioneine on the hepatic protein expression levels of Acetyl-FOXO1, FOXO1 and SIRT1. Results Compared with the model group, ergothioneine-treated groups decreased MDA and protein carbonyl levels in serum and liver, and increased SOD and CAT activities and GSH content. Western blotting indicated that ergothioneine administration reduced the ratio of Acetyl-FOXO1/FOXO1 and upregulated SIRT1 protein expression in liver tissue. Conclusion Ergothioneine exhibits significant protective effects against D-gal-induced oxidative damage in rats. The underlying mechanism may predominantly involve modulation of the FoxO signaling pathway.