摘要
目的探讨晚期糖基化终末产物(AGEs)对H9C2心肌细胞自噬的影响。方法用AGEs(100~400μg/mL)孵育H9C2心肌细胞24 h;应用MTT法检测心肌细胞活力,荧光显微镜观察GFP-LC3自噬小体的变化情况,Western blotting(WB)检测自噬微管相关蛋白1轻链3(LC3)、腺苷5′-单磷酸(AMP)活化蛋白激酶(AMPK)和磷酸化的AMPK(p-AMPK)的表达。并进一步运用AMPK激动剂(A-769662)和抑制剂(Compound C)预处理H9C2心肌细胞6 h后,运用AGEs(400μg/mL)孵育H9C2心肌细胞12 h,WB法检测LC3、p-AMPK、AMPK蛋白的表达。结果高浓度AGEs孵育H9C2心肌细胞24 h后降低细胞活力,并明显抑制GFP-LC3和LC3蛋白的表达(P<0.05),AMPK激动剂(A-769662)预处理可以逆转高浓度AGEs对H9C2心肌细胞p-AMPK的抑制作用,而AMPK抑制剂(Compound C)进一步降低p-AMPK的表达。结论高浓度AGEs抑制H9C2心肌细胞LC3表达,可能与抑制AMPK信号通路有关。
Objective To investigate the effects and mechanisms of advanced glycation end products(AGEs)on autophagy of H9 C2 cells.Methods H9 C2 cells were incubated with AGEs(100-400μg/mL)for 24 h.The cell viability was assessed by MTT;the change of GFP-LC3 signal was observed by fluorescence microscope;the expression of autophagy-related protein microtubule-associate protein 1 light chain3(LC3),adenosine 5’-monophosphate(AMP)-activated protein kinase(AMPK)and p-AMPK were detected by Western blotting(WB).H9 C2 cells were pretreated with A-769662(50μg/mL)and compound C(30μg/mL)for 6 h,and then incubated with AGEs(400μg/mL)for 12 h.WB was used to detect the expressions of LC3,p-AMPK and AMPK.Results The cell viability was reduced by high concentration of AGEs(P<0.05).GFP-LC3 puncta were significantly weakened by high concentration of AGEs(P<0.05).The expression of LC3 protein was significantly inhibited by high concentration of AGEs(P<0.05).Preconditioning with A-769662 reversed the inhibitory effect of AGEs(400μg/mL)on p-AMPK in H9 C2 cells,while compound C reduced the expression of p-AMPK.Conclusion High concentration of AGEs could inhibit the expression of LC3 in H9 C2 cells,which might be related to the inhibition of AMPK signaling pathway.
作者
何丽姗
梁梦
赖镇强
HE Li⁃shan;LIANG Meng;LAI Zhen⁃qiang(Clinical Pharmaceutics Room,Meizhou People's Hospital,Meizhou,Guangdong 514031;Gannan Medical University,Ganzhou,Jiangxi 341000;Pharmacy Intravenous Admixture Services,Meizhou People's Hospital,Meizhou,Guangdong 514031,China)
出处
《热带医学杂志》
CAS
2022年第1期52-55,60,共5页
Journal of Tropical Medicine
基金
广东省医学科学技术研究基金(B2021143)
梅州市人民医院科研培育项目(PY-C2019011)
