S1P通过调控S1PR1对高糖诱导的血管内皮细胞损伤的保护作用  

S1P Protects Against High Glucose-induced Vascular Endothelial Cell Injury Through S1PR1

作　　者：严惠 赵虎[2] Yan Hui;Zhao Hu(Department of Cardiology,Wuhan Fourth Hospital,Wuhan 430030,China;Division of Cardiology,Department of Internal Medicine,Tongji Hospital,Tongji Medical College,Huazhong University of Science and Technology,Wuhan 430030,China)

机构地区：[1]武汉市第四医院心血管内科,武汉430030 [2]华中科技大学同济医学院附属同济医院心血管内科,武汉430030

出　　处：《华中科技大学学报（医学版）》2024年第2期175-180,共6页Acta Medicinae Universitatis Scientiae et Technologiae Huazhong

基　　金：武汉市科学技术局知识创新专项项目(No.2022020801020562)。

摘　　要：目的探究1-磷酸鞘氨醇(sphingosine-1-phosphate,S1P)对高糖诱导血管内皮细胞损伤的作用及机制。方法利用高糖诱导人脐静脉内皮细胞(human umbilical vein endothelial cells,HUVECs)损伤,使用S1P及其1型受体(S1PR1)的小干扰RNA(si-S1PR1)处理细胞。采用qRT-PCR检测S1PR1转录水平以筛选高沉默效率的si-S1PR1用于后续实验,实验细胞分为对照组(NG)、高渗组(L-Glu)、高糖组(HG)、高糖+S1P组(HG+S1P)、高糖+si-NC+S1P组(HG+si-NC+S1P)、高糖+si-S1PR1+S1P组(HG+si-S1PR1+S1P)。应用CCK-8检测细胞增殖活力,划痕法检测细胞迁移能力,成管实验检测细胞血管形成能力,Western blot检测S1PR1及血管功能相关分子VEGF、VE-cadherin、eNOS的蛋白表达水平。结果与对照组相比,高糖组血管内皮细胞的增殖活力、迁移能力和血管形成能力均显著下降,并伴有S1PR1的表达降低(均P<0.01);同时高糖组细胞血管功能相关分子VEGF、VE-cadherin和eNOS的表达水平显著降低(均P<0.01);给予S1P干预后,S1PR1的表达水平及细胞活力、迁移和血管形成能力均显著回升,同时VEGF、VE-cadherin和eNOS蛋白表达水平亦显著增加(均P<0.01);而以si-S1PR1下调S1PR1表达则抵消了S1P对内皮细胞活力、迁移能力、血管形成能力及功能相关蛋白表达的恢复作用(均P<0.01)。结论S1P通过S1PR1上调血管功能相关分子VEGF、VE-cadherin、eNOS等的表达,对高糖环境下的血管内皮细胞发挥保护作用。Objective To explore the protective effect of sphingosine-1-phosphate(S1P)against high glucose-induced vascular endothelial cell injury,and its mechanism.Methods Human umbilical vein endothelial cell(HUVEC)injury was induced by high glucose,and the cells were treated with S1P and small interfering RNA(siRNA)of S1PR1.S1PR1 transcription level was detected by qRT-PCR to screen si-S1PR1 with high silencing efficiency for subsequent experiments.Then the cells were divided into control group(NG),hypertonic group(L-Glu),high glucose group(HG),high glucose+S1P group(HG+S1P),high glucose+si-NC+S1P group(HG+si-NC+S1P),high glucose+si-S1PR1+S1P group(HG+si-S1PR1+S1P).CCK-8 assay was used to detect the viability of endothelial cells.Then cell migration ability was detected by scratch method,and tube-forming experiment was conducted to detect the angiogenesis ability of endothelial cells.The protein expression levels of S1PR1 and vascular function-related molecules(VEGF,VE-cadherin and eNOS)were detected by Western blotting.Results Compared with control group,the viability,migration and angiogenesis ability of endothelial cells in the high glucose group were significantly decreased,while S1PR1 expression was reduced(all P<0.01).Meanwhile,the expression levels of vascular function-related molecules(VEGF,VE-cadherin and eNOS)were significantly downregulated(all P<0.01).After S1P treatment,S1PR1 expression and the viability,migration and angiogenesis ability of endothelial cells were significantly increased,and the protein expression levels of VEGF,VE-cadherin and eNOS were also increased significantly(P<0.01).Downregulation of S1PR1 by si-S1PR1 abolished the restoring effects of S1P on the viability,migration,angiogenesis ability of endothelial cells and the functionally related proteins(P<0.01).Conclusion S1P plays a protective role against high glucose induced injury of endothelial cells by upregulating VEGF,VE-cadherin and eNOS through S1PR1.

关 键 词：1-磷酸鞘氨醇 高糖 血管内皮细胞 血管功能 

分 类 号：R587.1[医药卫生—内分泌]