目的:结合生物信息学探讨瑞舒伐他汀(rosuvastatin，RST)对高糖诱导人脐静脉内皮细胞(human umbilical vein endothelial cells，HUVECs)损伤的影响及其作用机制。方法:网络药理学分析RST治疗糖尿病血管内皮功能障碍的作用靶点和信号通路，分子对接探索RST与核心靶点的结合能力。体外培养HUVECs，将细胞分为正常糖对照组、高糖组和高糖+RST(0。01、0。1、1、2、5和10 µmol/L)组，MTS法检测各组细胞活力;化学比色法检测HUVECs释放乳酸脱氢酶(lactate dehydrogenase，LDH)和一氧化氮(nitric oxide，NO)的情况;RT-qPCR法检测内皮型NO合成酶(endothelial NO synthase，eNOS)、密封蛋白1(claudin-1，CLDN-1)、闭合蛋白(occludin，OCLN)、闭锁小带蛋白1(zonula occludens-1，ZO-1)、醛糖还原酶(aldose reductase，AR)、Janus激酶2(Janus kinase 2，JAK2)、丝裂原活化蛋白激酶1(mitogen-activated protein kinase 1，MAPK1)、Ras同源家族成员A(Ras homologous gene family mem-ber A，RHOA)和热休克蛋白90AB1(heat shock proteins 90AB1，HSP90AB1)的mRNA表达变化;Western blot法检测细胞eNOS、OCLN和ZO-1蛋白表达水平。结果:网络药理学结果表明，RST可能通过作用于趋化因子信号通路、酪氨酸代谢、MAPK信号通路、缺氧诱导因子1α信号通路等缓解糖尿病性血管内皮功能障碍。MTS实验结果显示，RST显著提高高糖环境下HUVECs活力(P＜0。01)，并减轻高糖对HUVECs的损伤。与正常糖对照组比较，高糖组中细胞释放LDH水平显著升高(P＜0。01)，NO、eNOS、CLDN-1、OCLN和ZO-1的表达水平显著降低(P＜0。05);且高糖组中AR、JAK2、MAPK1和RHOA的mRNA表达水平升高(P＜0。01)，HSP90AB1的mRNA表达水平下降(P＜0。05)。而与高糖组对比，高糖+RST组中LDH水平显著下降(P＜0。01)，NO、eNOS、CLDN-1、OCLN和ZO-1的表达水平显著升高(P＜0。01);且高糖+RST组中AR、JAK2、MAPK1和RHOA的mRNA表达水平均降低(P＜0。01)，而HSP90AB1 mRNA表达水平升高(P＜0。01)。结论:RST可减轻高糖诱导的血管内皮损伤，其机制可能与RST抑制AR、JAK2、MAPK1和RHOA表达，促进HSP90AB1表达有关。
Mechanism of rosuvastatin attenuating high glucose-induced vascular en-dothelial injury based on bioinformatics
AIM:This study investigated the protective effects and underlying mechanisms of rosuvastatin(RST)in mitigating high glucose(HG)-induced damage in human umbilical vein endothelial cells(HUVECs),comple-mented by bioinformatics analysis.METHODS:Network pharmacology was employed to identify the potential targets and signaling pathways of RST in treating HG-induced vascular endothelial dysfunction.Molecular docking techniques were used to evaluate the binding affinity of RST to these core targets.The HUVECs were cultured in vitro and assigned into control,HG,and HG+RST(0.01,0.1,1,2,5 and 10 µmol/L)groups.Cell viability was determined using the MTS as-say.Levels of lactate dehydrogenase(LDH)and nitric oxide(NO)were quantified using chemical colorimetric assays.The mRNA levels of endothelial nitric oxide synthase(eNOS),claudin-1(CLDN-1),occludin(OCLN),zonula oc-cludens-1(ZO-1),aldose reductase(AR),Janus kinase 2(JAK2),mitogen-activated protein kinase 1(MAPK1),Ras homologous gene family member A(RHOA)and heat shock proteins 90AB1(HSP90AB1)were assessed by RT-qPCR.Western blot analysis was used to evaluate protein levels of eNOS,OCLN and ZO-1.RESULTS:Network pharmacology analysis suggested that RST may improve HG-induced vascular endothelial dysfunction by influencing the chemokine sig-naling pathway,tyrosine metabolism,MAPK signaling pathway,and hypoxia-inducible factor 1 alpha signaling pathway.The MTS assay indicated that RST significantly enhanced cell viability in an HG environment(P＜0.01)and reduced HG-induced damage in HUVECs.Compared with the control group,the HG group showed a significant increase in LDH levels(P＜0.01)and decreases in NO,eNOS,CLDN-1,OCLN and ZO-1 levels(P＜0.05).Additionally,the mRNA levels of AR,JAK2,MAPK1 and RHOA were elevated(P＜0.01),and HSP90AB1 was reduced in the HG group(P＜0.05).Rel-ative to the HG group,RST treatment significantly decreased LDH levels(P＜0.01)and increased the levels of NO,eNOS,CLDN-1,OCLN and ZO-1(P＜0.01).Moreover,the mRNA levels of AR,JAK2,MAPK1 and RHOA were re-duced(P＜0.01),and HSP90AB1 expression was increased in the HG+RST group(P＜0.01).CONCLUSION:RST ef-fectively attenuates HG-induced endothelial injury.This protective effect is potentially mediated by downregulating AR,JAK2,MAPK1,and RHOA expression and upregulating HSP90AB1 expression.
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