长链非编码RNA HIF1α-AS1在血管内皮细胞缺氧损伤中的作用研究  

The study on long non-coding RNA hypoxia-inducible factor-1α-antisense RNA1 in hypoxic injury of vascular endothelial cells

作　　者：王微 王岩 王超君 卢芳芳 凌静 WANG Wei;WANG Yan;WANG Chao-jun;LU Fang-fang;LING Jing(Department of Pharmacy and Equipment,Air Force Health Care Center for Special Services Hangzhou Area 1,Hangzhou 310007,Zhejiang,China;College of Animal Science and Technology,Zhejiang Forest University,Hangzhou 311300,Zhejiang,China)

机构地区：[1]空军杭州特勤疗养中心疗养一区药械科,杭州310007 [2]浙江农林大学动物科技学院动物医学院,杭州311300

出　　处：《东南国防医药》2022年第1期1-5,共5页Military Medical Journal of Southeast China

基　　金：杭州市卫生科技计划项目(2014B01)。

摘　　要：目的检测长链非编码RNA缺氧诱导因子-1alpha-反义链1(lncRNA HIF-1α-AS1)在缺糖缺氧血管内皮细胞中的表达及对细胞缺氧应激损伤的影响。方法原代培养人脐静脉血管内皮细胞并建立缺糖缺氧模型,实时荧光定量聚合酶链式反应(Real-time PCR)方法从95种长链非编码RNA中筛选出缺糖缺氧后表达差异显著的基因lncRNA HIF-1α-AS1。用流式细胞仪、TUNEL/Hochest检测细胞凋亡情况。siRNA干扰lncRNA HIF1α-AS1表达后,通过乳酸脱氢酶漏出率、Western blot检测超氧化物歧化酶1(SOD1)表达等方法评估细胞损伤情况。结果血管内皮细胞建立缺糖缺氧模型后,lncRNA HIF-1α-AS1含量显著上升1.98倍(P<0.05)。对lncRNA HIF-1α-AS1进行干扰后,3个干扰组细胞乳酸脱氢酶(LDH)漏出率与模型组相比显著下降,siRNA1组为20.84%±2.64%(P<0.05),siRNA2组为19.82%±1.61%(P<0.01),siRNA3组为17.01%±0.24%(P<0.01)。siRNA2和siRNA3的SOD1基因mRNA水平显著上调(P<0.01);2组SOD1蛋白表达量也明显上升,siRNA2(P<0.05),siRNA3(P<0.01),细胞损伤减轻。结论干扰lncRNA HIF-1α-AS1的表达能够减轻血管内皮细胞缺糖缺氧后的损伤,提高细胞的抗缺氧能力,这可能与SOD1表达量升高、乳酸脱氢酶漏出减少有关。Objective This study aimed to explore the mechanism of long non-coding RNA hypoxia-inducible factor-1alpha-antisense RNA1(lncRNA HIF-1α-AS1)in vascular endothelial cell hypoxia stress injury.Methods In this study,the primary human umbilical vein endothelial cells(HUVEC)were cultured and oxygen-glucose deprivation model was established.The lncRNA HIF-1α-AS1 was significantly increasing screened from 95 lncRNAs measured by Real-time PCR(RT-PCR).After RNA interference,anti-hypoxia stress of the cell ability was detected by the lactate dehydrogenase leakage rate and Western detection of superoxide dismutase1(SOD1)expression in vitro to explore the specific role of this lncRNA in HUVEC.Results The expression of lncRNA HIF-1α-AS1 was significantly increased by 1.98 times(P<0.05)after oxygen-glucose deprivation(OGD).After lncRNA HIF-1α-AS1 interference,the leakage rate of lactate dehydrogenase(LDH)was significantly decreased,20.84%±2.64%(P<0.05),19.82%±1.61%(P<0.01)and 17.01%±0.24%(P<0.01)in siRNA1 group,siRNA2 group and siRNA3 group,respectively.The mRNA levels of SOD1 gene in siRNA2 group and siRNA3 group were significantly up-regulated(P<0.01).The expression of SOD1 protein was also significantly increased in the two groups.The cell damage was reduced in siRNA2 group(P<0.05)and siRNA3 group(P<0.01).Conclusion Inhibition of lncRNA HIF-1α-AS1 expression can reduce the stress damage of HUVEC after hypoxia,and improve the anti-hypoxia ability of cells,which may be related to the increase of SOD1 activity and the decrease of LDH leakage.

关 键 词：长链非编码RNA 缺氧诱导因子 缺氧应激损伤 血管内皮细胞 

分 类 号：R543.6[医药卫生—心血管疾病]