miR-103a-3p通过p38 MAPK信号通路抑制高糖诱导的血管内皮细胞损伤  

miR-103a-3p inhibits high glucose-induced vascular endothelial cell injury through p38 MAPK signaling pathway

作　　者：班维固[1] 齐辉[1] 卿鹏[2] 滕秀英[1] 陆丽娜 许文婷 丛宏政 史美超 BAN Wei-gu;QI Hui;QING Peng;TENG Xiu-ying;LU Li-na;XU Wen-ting;CONG Hong-zheng;SHI Mei-chao(Second Department of Rehabilitation Medicine,The Second Affiliated Hospital of Heilongjiang University of Chinese Medicine,Harbin 150001,Heilongjiang Province,China;Department of Acupuncture and Moxibustion,The First Affiliated Hospital of Jinan University,Guangzhou 510630,Guangdong Province,China;Graduate School,Heilongjiang University of Traditional Chinese Medicine,Harbin 150040,Heilongjiang Province,China)

机构地区：[1]黑龙江中医药大学附属第二医院康复医学二科,黑龙江哈尔滨150001 [2]暨南大学附属第一医院针灸科,广东广州510630 [3]黑龙江中医药大学研究生学院,黑龙江哈尔滨150040

出　　处：《中国临床药理学杂志》2022年第21期2554-2558,共5页The Chinese Journal of Clinical Pharmacology

摘　　要：目的探讨miR-103a-3p对高糖(HG)诱导的血管内皮细胞损伤的影响,分析其机制是否与调控p38丝裂原活化蛋白激酶(MAPK)信号通路有关。方法将血管内皮细胞(HUVECs)分为NC组(用含5 mmol·L^(-1)葡萄糖培养液)、HG组(含33 mmol·L^(-1)葡萄糖培养液)、miR-NC+HG组(转染miR-NC+33 mmol·L^(-1)葡萄糖培养液)、miR-103a-3p+HG组(转染miR-103a-3p+33 mmol·L^(-1)葡萄糖培养液)、miR-103a-3p+Anisomycin+HG组(转染miR-103a-3p+33 mmol·L^(-1)葡萄糖培养液+10μmol·L^(-1)的茴香霉素)。分别使用细胞计数试剂盒-8(CCK-8)法、实时荧光定量聚合酶链反应(RT-qPCR)、流式细胞术检测HUVECs细胞活力、miR-103a-3p表达以及细胞凋亡率。比色法检测超氧化物歧化酶(SOD)活性、丙二醛(MDA)含量以及乳酸脱氢酶(LDH)释放量。蛋白质印迹法检测磷酸化的p38 MAPK(p-p38)蛋白表达。结果NC组、HG组、miR-NC+HG组、miR-103a-3p+HG组、miR-103a-3p+Anisomycin+HG组HUVECs活力为1.04±0.07,0.55±0.03,0.55±0.05,0.95±0.07,0.54±0.05;细胞凋亡率为(4.46±0.32)%,(20.29±1.35)%,(20.19±1.26)%,(7.13±0.61)%,(24.36±1.36)%;SOD活性为(15.22±1.02),(8.79±0.75),(8.71±0.57),(13.99±0.60)和(7.94±0.67)U·mL^(-1);MDA含量为(1.06±0.09),(4.99±0.29),(4.94±0.30),(1.75±0.12)和(4.97±0.23)μmol·L^(-1);LDH释放量为(623.13±26.32),(1197.10±76.95),(1207.11±65.19),(751.90±42.04)和(1225.90±68.73)U·L^(-1);p-p38蛋白水平为0.34±0.03,0.79±0.08,0.82±0.06,0.40±0.05和0.86±0.06。上述指标,NC组与HG组比较、miR-NC+HG组与miR-103a-3p+HG组比较、miR-103a-3p+HG组与miR-103a-3p+Anisomycin+HG组比较,差异均具有统计学意义(均P<0.05)。结论miR-103a-3p可通过抑制p38 MAPK信号通路减轻高糖诱导的血管内皮细胞损伤。Objective To evaluate the effects of miR-103 a-3 p on high glucose(HG)-induced vascular endothelial cell injury,and further investigate whether its mechanism is related to the regulation of p38 mitogen-activated protein kinase(MAPK)signaling pathway.Methods Human umbilical vein vascular endothlial cells(HUVECs)cells were divided into the control(NC)group(medium containing 5 mmol·L^(-1) glucose),HG group(33 mmol·L^(-1) glucose culture medium),miR-NC+HG group(33 mmol·L^(-1)glucose culture medium after transfection with miR-NC),miR-103a-3p+HG group(33 mmol·L^(-1)glucose culture medium after transfection with miR-103a-3p),and miR-103a-3p+anisomycin+HG group(33 mmol·L^(-1)glucose medium and 10μmol·L^(-1)anisomycin after transfection with miR-103a-3p).The cell viability,miR-103a-3pexpression and apoptosis rate of human umbilical vein endothelial cells(HUVECs)were detected by cell counting kit-8(CCK-8)assay,real-time fluorescence quantitative polymerase chain reaction(RT-qPCR)and flow cytometry.The superoxide dismutase(SOD)activity,malondialdehyde(MDA)content and lactate dehydrogenase(LDH)release were detected by colorimetric method.The expression of phosphorylated p38 MAPK(p-p38)proteins was detected by Western blotting.Results The cell activity of HUVECs in NC group,HG group,miR-NC+HG group,miR-103a-3p+HG group,miR-103a-3p+Anisomycin+HG group were 1.04±0.07,0.55±0.03,0.55±0.05,0.95±0.07,0.54±0.05;apoptosis rates were(4.46±0.32)%,(20.29±1.35)%,(20.19±1.26)%,(7.13±0.61)%,(24.36±1.36)%;SOD activity were(15.22±1.02),(8.79±0.75),(8.71±0.57),(13.99±0.60),(7.94±0.67)U·mL^(-1);MDA content were(1.06±0.09),(4.99±0.29),(4.94±0.30),(1.75±0.12),(4.97±0.23)μmol·L^(-1);LDH release were(623.13±26.32),(1197.10±76.95),(1207.11±65.19),(751.90±42.04),(1225.90±68.73)U·L^(-1);p-p38 protein levels were 0.34±0.03,0.79±0.08,0.82±0.06,0.40±0.05,0.86±0.06.The differences in the above indicators were statistically significant between the NC group and the HG group,between the miR-NC+HG group and the miR-10

关 键 词：微小RNA-103a-3p 血管内皮细胞 高糖 凋亡 氧化应激 p38丝裂原活化蛋白激酶信号通路 

分 类 号：R97[医药卫生—药品]