松茸多肽调控AMPK/Sirt1/PGC1ɑ通路保护多柔比星诱导心肌损伤

Tricholoma matsutake peptides protects from doxorubicin-induced cardiotoxicity through AMPK/Sirt1/PGC1ɑ pathways

目的:松茸是一种珍贵的食用菌和药用真菌,可从中分离纯化出血管紧张素转换酶(ACE)抑制肽,已被证实对自发性高血压或其他心血管疾病有益。本文将探讨松茸多肽(tricholoma matsutake polypeptide,TMP)对多柔比星(Dox)诱导的心肌损伤的保护作用和机制。方法:分组培养大鼠心肌细胞H9c2,分别给药24 h,CCK8法检测细胞存活率筛选出Dox、TMP、右雷佐生(Dexra)的最佳联合给药浓度,检测丙二醛(MDA)、超氧化物歧化酶(SOD)、烟酰胺腺嘌呤二核苷酸(NAD+)/烟酰胺腺嘌呤二核苷酸(NADH)、ATP、乳酸脱氢酶(LDH)含量变化,使用荧光显微镜观察JC-1线粒体膜电位和TUNEL细胞凋亡染色情况,Annexin V-FITC/PI双染细胞检测细胞凋亡情况,β-半乳糖染色实验观察细胞衰老情况,Western blot法检测相关蛋白表达。结果:TMP可显著改善Dox诱导的细胞存活率下降,且能有效抑制Dox诱导的细胞衰老、氧化应激损伤、线粒体功能损伤和细胞凋亡。Western blot结果显示,TMP可能通过抑制Dox诱导的过度自噬和AMPK/Sirt1/PGC1ɑ通路激活能量代谢,保护H9c2免受Dox诱导的心肌损伤。结论:TMP对Dox诱导的心肌损伤有保护作用。这种保护伴随着自噬流稳态的恢复,增加腺苷酸活化蛋白激酶(AMPK)依赖的能量代谢,降低氧化应激,改善线粒体功能。

Objective: Tricholoma matsutake is a precious edible and medicinal fungus,from which angiotensin-converting enzyme inhibitory peptides can be isolated and purified,and it has been proved to be beneficial to spontaneous hypertension or other cardiovascular diseases. This article will explore the protective effect and mechanism of matsutake polypeptide on doxorubicin-induced myocardial injury. Methods: The rat cardiomyocytes H9c2 were cultured in groups and administered with drugs for 24 h respectively. The cell viability was detected by the CCK8 method to screen out the best-combined administration concentration of Dox,TMP,and dexrazoxane.MDA,SOD,NAD +/NADH,ATP,LDH contents were detected,JC-1 mitochondrial membrane potential and TUNEL cell apoptosis staining were observed by fluorescence microscope,cell apoptosis was detected by Annexin V-FITC/PI double-stained cells,and cell senescence was observed by β-galactose staining experiment,and Western blotting was used to detect the expression of related proteins. Results: It was found that 30 μg·m L-1TMP could significantly improve Dox-induced cell viability decline and effectively inhibit Dox-induced cell senescence,oxidative stress damage,mitochondrial function damage,and cellular apoptosis. Western blotting results showed that TMP may activate energy metabolism by inhibiting Dox-induced excessive autophagy and AMPK/Sirt1/PGC1ɑ pathway and protect H9c2 from Dox-induced myocardial injury. Conclusion: TMP has a protective effect on Dox-induced myocardial injury. This protection is accompanied by restoration of autophagic flux homeostasis,increased AMPKdependent energy metabolism,decreased oxidative stress,and improved mitochondrial function.