摘要
目的:探讨小核仁RNA宿主基因3(SNHG3)对过氧化氢(H_(2)O_(2))处理的血管内皮细胞损伤的影响及分子机制。方法:将Eahy926细胞分为对照组(不做任何处理)、H_(2)O_(2)组(500 mmol/L H_(2)O_(2))、H_(2)O_(2)+pcDNA3.1组(转染pcDNA3.1+500 mmol/L H_(2)O_(2))、H_(2)O_(2)+pcDNA3.1-SNHG3组(转染pcDNA3.1-SNHG3+500 mmol/L H_(2)O_(2))、H_(2)O_(2)+pcDNA3.1-SNHG3+miR-NC组(转染pc-DNA3.1-SNHG3和miR-NC+500 mmol/L H_(2)O_(2))、H_(2)O_(2)+pcDNA3.1-SNHG3+miR-186组(转染pcDNA3.1-SNHG3和miR-NC+500 mmol/L H_(2)O_(2))。RT-qPCR检测SNHG3和miR-186表达;四甲基偶氮唑盐比色法(MTT)检测细胞存活率;超氧化物歧化酶(SOD)和乳酸脱氢酶(LDH)试剂盒分别检测细胞SOD活性和培养液LDH含量;流式细胞术检测细胞凋亡;荧光素酶报告实验检测SNHG3和miR-186的靶向关系。结果:与对照组相比,H_(2)O_(2)处理的Eahy926细胞中SNHG3表达、存活率、SOD活性显著降低,LDH水平、细胞凋亡率显著升高(P<0.05)。过表达SNHG3,H_(2)O_(2)处理的Eahy926细胞存活率、SOD活性显著升高,LDH水平、细胞凋亡率显著降低(P<0.05)。SNHG3靶向调控miR-186表达,过表达miR-186逆转了SNHG3对H_(2)O_(2)处理的Eahy926细胞活性、凋亡及SOD、LDH的影响。结论:过表达SNHG3可促进H_(2)O_(2)处理的Eahy926细胞存活,抑制细胞凋亡,提高SOD活性,降低LDH水平,对H_(2)O_(2)诱导的Eahy926细胞损伤具有保护作用,其机制可能与miR-186有关。
Objective:To investigate effect and molecular mechanism of small nucleolar RNA host gene 3(SNHG3)on vascular endothelial cell injury induced by hydrogen peroxide(H_(2)O_(2)).Methods:Eahy926 cells were divided into control group(without any treatment),H_(2)O_(2)group(500 mmol/L H_(2)O_(2)),H_(2)O_(2)+pcDNA3.1 group(transfected with pcDNA3.1+500 mmol/L H_(2)O_(2)),H_(2)O_(2)+pcDNA3.1-SNHG3 group(transfected with pcDNA3.1-SNHG3+500 mmol/L H_(2)O_(2)),H_(2)O_(2)+pcDNA3.1-SNHG3+miR-NC group(transfected with pcDNA3.1-SNHG3 and miR-NC+500 mmol/L H_(2)O_(2)),H_(2)O_(2)+pcDNA3.1-SNHG3+miR-186 group(transfected with pcDNA3.1-SNHG3 and miR-NC+500 mmol/L H_(2)O_(2)).RT-qPCR was used to detect expressions of SNHG3 and miR-186;tetramethylazozolium salt colorimetric assay(MTT)was used to detect survival rate;superoxide dismutase(SOD)and lactate dehydrogenase(LDH)kits were used to detect SOD activity and LDH content in culture medium;cell apoptosis was detected by flow cytometry;luciferase reporter assay was used to detect targeting relationship between SNHG3 and miR-186.Results:Compared with control group,expression of SNHG3,survival rate and SOD activity in H_(2)O_(2)-treated Eahy926 cells were significantly decreased,LDH level and apoptosis rate were significantly increased(P<0.05).Overexpression of SNHG3,cell survival rate,SOD activity were significantly increased,and LDH level and apoptosis rate were significantly decreased(P<0.05).SNHG3 targeting regulates miR-186 expression.Overexpression of miR-186 reversed effect of SNHG3 on H_(2)O_(2)-treated Eahy926 cell activity,apoptosis,SOD and LDH.Conclusion:Overexpression of SNHG3 can promote survival of H_(2)O_(2)-treated Eahy926 cells,inhibit cell apoptosis,increase SOD activity,decrease LDH level,and protect H_(2)O_(2)-induced Eahy926 cell injury,whose mechanism may be related to miR-186.
作者
郑婉
林云
左琦
林燕仔
颜亚妮
符碧薇
ZHENG Wan;LIN Yun;ZUO Qi;LIN Yanzai;YAN Yani;FU Biwei(Department of Cardiovascular Medicine,the First Affiliated Hospital of Hainan Medical University,Haikou 570102,China)
出处
《中国免疫学杂志》
CAS
CSCD
北大核心
2022年第19期2330-2335,共6页
Chinese Journal of Immunology
