LncRNA SNHG10对脂多糖诱导血管内皮细胞氧化应激损伤的影响  

Effect of lncRNA SNHG10 on LPS-induced oxidative stress injury of vascular endothelial cells

作　　者：修晟尧 郭楠[2] 凌书策 肖珉[1] 常佩芬[1] XIU Shengyao;GUO Nan;LING Shuce;XIAO Min;CHANG Peifen(Department of Cardiology,Dongzhimen Hospital,Beijing University of Traditional Chinese Medicine,Beijing 100010,China;不详)

机构地区：[1]北京中医药大学东直门医院心血管科,北京100010 [2]北京中医药大学东直门医院ICU

出　　处：《山东医药》2023年第28期41-45,共5页Shandong Medical Journal

摘　　要：目的探讨lncRNA SNHG10对脂多糖(LPS)诱导血管内皮细胞氧化应激损伤的影响。方法通过筛选出的LPS最适浓度500 ng/mL诱导人脐静脉内皮细胞HUVEC,构建血管内皮细胞氧化应激损伤体外模型。取对数生长期的HUVEC细胞,分别将si-NC、si-SNHG10质粒转染至HUVEC细胞,采用LPS(500 ng/mL)处理(分别记为LPS+si-NC组、LPS+si-SNHG10组),选择未行任何处理的HUVEC细胞为Control组,仅加入500 ng/mL LPS处理的细胞为LPS组。CCK-8法、流式细胞术检测细胞活力和凋亡水平;RT-qPCR法检测小核仁RNA宿主基因10(SNHG10)相对表达量;Western blotting法检测Ki-67、Bcl-2和Bax蛋白表达;ELISA法检测TNF-α、IL-6、IL-1β、MDA含量和SOD活性。结果LPS+si-SNHG10组HUVEC细胞活力、Ki-67蛋白、Bcl-2蛋白表达分别为(75.6±4.1)%、0.74±0.06、0.65±0.05,LPS+si-NC组分别为(45.4±2.8)%、0.52±0.04、0.42±0.03;LPS+si-SNHG10组SNHG10表达、细胞凋亡率、Bax蛋白表达、TNF-α、1L-6和1L-1β含量分别为1.88±0.11、(16.8±0.9)%、1.32±0.07、(289.2±25.1)pg/mL、(363.3±22.4)pg/mL、(402.6±21.3)pg/mL,LPS+si-NC组分别为3.24±0.19、(23.1±1.2)%、1.68±0.11、(536.6±33.0)pg/mL、(634.5±33.1)pg/mL、(664.5±38.1)pg/mL,LPS+si-SNHG10组SNHG10表达、细胞凋亡率、Bax蛋白表达、TNF-α、1L-6和1L-1β含量低于LPS+si-NC组(P均<0.05)。与Control组相比,LPS组HUVEC细胞内MDA表达升高(P<0.05),SOD活性降低(P<0.05);与LPS+si-NC组相比,LPS+si-SNHG10组HUVEC培养液MDA含量降低(P<0.05),SOD活性升高(P<0.05)。结论干扰SNHG10表达可以减轻LPS诱导的HUVEC凋亡、炎症和氧化应激,促进细胞增殖。Objective To explore the effect of lncRNA SNHG10 on lipopolysaccharide(LPS)-induced oxidative stress injury in vascular endothelial cells.Methods The optimal concentration of LPS was screened out to be 500 ng/mL.The in vitro models were constructed by inducing human umbilical vein endothelial cells(HUVEC)with LPS.HUVEC in the logarithmic growth phase were taken,si-NC and si-SNHG10 plasmids were transfected into HUVEC,and then the cells were treated with LPS(500 ng/mL)(recorded as LPS+si-NC group and LPS+si-SNHG10 group,respectively).HUVEC without any treatment were selected as the Control group,and only 500 ng/mL LPS was added.CCK-8 and flow cytometry were used to detect cell viability and apoptosis level;RT-qPCR was used to detect the relative expression of small nucleolar RNA host gene 10(SNHG10);Western blotting was used to detect Ki-67,Bcl-2,and Bax;ELISA was used to detect tumor necrosis factor-α(TNF-α),interleukin 6(IL-6),interleukin 1β(IL-1β),MDA content,and SOD activity.Results The cell viability,Ki-67 protein,and Bcl-2 protein expression of HUVEC in the LPS+si-SNHG10 group were 75.6%±4.1%,0.74±0.06,0.65±0.05,and they were 45.4±2.8%,0.52%±0.04,and 0.42±0.03,respectively,in the LPS+si-NC group;SNHG10 expression,apoptosis rate,Bax protein expression,TNF-α,1L-6 and 1L-1βexpression in the LPS+si-SNHG10 group were 1.88±0.11,16.8%±0.9%,1.32±0.07,(289.2±25.1)pg/mL,(363.3±22.4)pg/mL,and(402.6±21.3)pg/mL,which were 3.24±0.19,23.1%±1.2%,1.68±0.11,(536.6±33.0)pg/mL,(634.5±33.1)pg/mL,and(664.5±38.1)pg/mL,respectively,in the LPS+si-NC group.SNHG10 expression,apoptosis rate,Bax protein expression,TNF-α,1L-6,and 1L-1βexpression levels were lower in the LPS+si-SNHG10 group than in the LPS+si-NC group(all P<0.05).Compared with the Control group,the expression of MDA increased in the LPS group(P<0.05),and the SOD activity decreased(P<0.05);compared with the LPS+si-NC group,MDA in HUVEC cells of the LPS+si-SNHG10 group decreased(P<0.05),while the SOD activity increased(P<0.05).Conclusion Interferin

关 键 词：炎症反应 小核仁RNA宿主基因10 人脐静脉内皮细胞 氧化应激 脂多糖 

分 类 号：R364.5[医药卫生—病理学]