摘要
目的:研究杨梅素(myricetin,MYR)对氧糖剥夺/再灌注(oxygen-glucose deprivation/reperfusion,OGD/R)诱导的人心肌细胞损伤的保护作用,并探讨其对低氧诱导因子(hypoxia-inducible factor,HIF)信号通路的影响。方法:体外培养人心肌细胞AC16,并将其随机分为对照组、模型组和MYR处理组;对照组给予正常培养基常规培养,模型组进行OGD/R处理,MYR处理组即在模型组添加MYR;利用低氧细胞培养系统构建OGD/R模型;CCK-8法检测细胞生存;流式细胞术检测细胞凋亡、线粒体膜电位(mitochondrial membrane potential,MMP)和活性氧(reactive oxygen species,ROS)含量;使用过氧化氢酶(catalase,CAT)活性检测试剂盒检测AC16细胞内CAT活性;通过蛋白质免疫印迹法检测HIF通路相关蛋白的表达。结果:氧糖剥夺18 h/复氧4 h(OGD_(18 h)/R_(4 h))为合适造模条件,此时AC16细胞生存率为(60.26±3.01)%;5、10、20、30、40μmol·L^(–1)MYR均可以显著逆转OGD_(18 h)/R_(4 h)导致的细胞损伤,其中10、20μmol·L^(–1)MYR处理组,细胞生存率分别为(77.39±2.17)%和(82.80±5.58)%,与单纯模型组相比,差异最为显著(P<0.01);与模型组相比,10、20μmol·L^(–1)MYR处理组的细胞凋亡率、低MMP的细胞比率、ROS水平以及HIF-1α、p-Akt、Bax和Bcl-2蛋白表达水平均显著下降(均P<0.01);与模型组相比,10、20μmol·L^(–1)MYR处理组CAT活性显著增加(P<0.01)。结论:MYR通过提高CAT活性,降低OGD/R诱导的ROS水平,发挥心肌细胞保护作用,并限制HIF信号通路的应激激活。
OBJECTIVE To evaluate the protective effects of myricetin(MYR)against oxygen-glucose deprivation/reperfusion(OGD/R)-induced injury in human cardiomyocytes and to investigate its impact on the hypoxia-inducible factor(HIF)signaling pathway.METHODS Human AC16 cardiomyocytes were cultured and randomly assigned to control,model(OGD/R),and MYR treatment(OGD/R+MYR)groups.The OGD/R model was established using a hypoxic cell culture system.Cell viability was measured by CCK-8 assay.Apoptosis,mitochondrial membrane potential(MMP),and reactive oxygen species(ROS)were assessed by flow cytometry.Catalase(CAT)activity was measured with a commercial detection kit.Expression of proteins related to the HIF pathway and apoptosis was analyzed by Western blotting.RESULTS OGD for 18 hours followed by reoxygenation for 4 hours(OGD_(18 h)/R_(4 h))reduced AC16 cell viability to(60.26±3.01)%,and was selected as the optimal injury model.MYR at 5,10,20,30,and 40μmol·L^(–1)significantly attenuated OGD_(18 h)/R_(4 h)-induced cell injury;the 10 and 20μmol–1 doses produced the greatest protection,restoring viability to(77.39±2.17)%and(82.80±5.58)%,respectively(both P<0.01 vs.model).Compared with the model group,treatment with 10 and 20μmol·L^(–1)MYR significantly reduced apoptosis rate,the proportion of cells with decreased MMP,ROS levels,and the expression of HIF-1alpha(HIF-1α),p-protein kinase B(Akt),Bax,and Bcl-2(all P<0.01).MYR(10 and 20μmol·L^(–1))also markedly increased CAT activity relative to the model group(P<0.01).CONCLUSION MYR protects human cardiomyocytes from OGD/R-induced injury,likely by increasing CAT activity,reducing ROS accumulation,and attenuating stress-induced activation of the HIF signaling pathway.
作者
何孟瑜
苏楠
蒋思齐
刘改枝
马永成
HE Mengyu;SU Nan;JIANG Siqi;LIU Gaizhi;MA Yongcheng(School of Pharmacy,Henan University of Chinese Medicine,Henan Zhengzhou 450046,China;College of Food and Biological Engineering,Henan University of Animal Husbandry and Economy,Henan Zhengzhou 450011,China;School of Pharmacy,Zhengzhou University,Henan Zhengzhou 450001,China;Henan Key Laboratory of Individualized Drug Therapy for Cardiovascular Diseases,Centeral China Fuwai Hospital of Zhengzhou University,Henan Zhengzhou 451464,China)
出处
《中国医院药学杂志》
北大核心
2026年第1期35-41,共7页
Chinese Journal of Hospital Pharmacy
基金
河南省自然科学基金项目(编号:222300420573)
河南省医学科技攻关项目(编号:LHGJ20240158)
国家自然科学基金资助项目(编号:81502952)
河南省高等学校重点科研项目(编号:25A350007)。
关键词
杨梅素
缺血再灌注损伤
缺氧诱导因子
氧化应激
myricetin
ischemia-reperfusion injury
hypoxia-inducible factor
oxidative stress
