二氢杨梅素对高糖刺激下心肌细胞坏死性凋亡的影响
The effects of dihydromyricetin on necroptosis in high glucose-stimulated cardiomyocytes
孙琳琳 1施益金 1施卉 1陈万杰 1顾锦华1
作者信息
- 1. 226007 南通,南通大学附属妇幼保健院药学部
- 折叠
摘要
目的 探究二氢杨梅素(DHY)对高糖刺激下心肌细胞损伤的影响.方法 原代心肌细胞用DHY(80 μmol/L)预处理4 h,用正常葡萄糖(NG)5.5 mmol/L或高糖(HG)33.3 mmol/L刺激48 h.随后利用MTT法评估细胞活性,测定乳酸脱氢酶(LDH)评估细胞损伤程度,二氢乙锭(DHE)和MitoSOX染色评估氧化应激水平,蛋白印迹和免疫荧光检测受体相互作用蛋白激酶3(RIPK3)蛋白表达,原位末端转移酶标记(TUNEL)法检测心肌细胞凋亡程度.结果 与HG组相比,DHY显著增强高糖刺激下心肌细胞活性(P=0.003),减少LDH释放(P=0.001),减弱DHE和MitoSOX染色荧光强度,显著抑制RIPK3蛋白表达(P＜0.01),减少TUNEL染色阳性细胞数量(P＜0.01).结论 DHY增强高糖刺激下心肌细胞活性,减轻损伤程度,可能与DHY抑制氧化应激和减弱坏死性凋亡有关.
Abstract
Objective To investigate the effects of dihydromyricetin in high glucose-stimulated cardiomyocytes injury.Methods Primary cardiomyocytes were pretreated with DHY(80 μmol/L)for 4 h,followed by normal glucose(NG,5.5 mmol/L)or high glucose(HG,33.3 mmol/L)stimulation for 48 h.Subsequently,cell viability was assessed by MTT.The degree of cell damage was evaluated by lactate dehydrogenase(LDH)measurement.The level of oxidative stress was assessed with dihydroethidium(DHE)and MitoSOX staining.The protein expression of receptor interacting protein kinase 3(RIPK3)was detected by immunofluorescence and Western blot.The degree of cardiomyocytes apoptosis was determined by TdT-mediated dUTP nick end labeling(TUNEL).Results Compared with HG group,DHY significantly enhanced cardiomyocytes activity(P=0.003),reduced LDH release(P=0.001),alleviat-ed the fluorescence intensity of DHE and MitoSOX staining,inhibited RIPK3 protein expression(P＜0.01),and reduced the number of positive cells with TUNEL staining(P＜0.01)in high glucose-stimulated cardiomyocytes.Conclusion DHY increased cardiomyo-cytes viability and attenuated cell injury,which might be related to oxidative stress inhibition and necroptosis attenuating in high glu-cose-stimulated cardiomyocytes.
关键词
二氢杨梅素/高糖/氧化应激/受体相互作用蛋白激酶3Key words
dihydromyricetin/high glucose/oxidative stress/receptor interacting protein kinase 3引用本文复制引用
基金项目
南通市科技计划(MSZ2022097)
南通市卫生健康委科研项目(QNZ2023072)
出版年
2024
NETL
NSTL
万方数据