Cinaciguat对高糖损伤血管内皮细胞功能的影响

Effects of cinaciguat on function of human umbilical vein endothelial cells injured by high glucose

目的糖尿病状态下,一氧化氮(NO)与可溶性鸟苷酸环化酶(soluble guanylatecyclase,sGC)均被氧化,使NO-sGC-环磷酸鸟苷(cGMP)信号通路传导异常,NO-sGC-cGMP信号通路异常是糖尿病血管内皮功能障碍的重要病理基础。可溶性鸟苷酸环化酶激活剂(cinaciguat,CIN)可激活失活的sGC进而起到保护血管内皮细胞功能的作用。本研究通过体外实验制造高糖内皮细胞损伤模型,观察CIN对人脐静脉内皮细胞功能的影响。方法培养人脐静脉内皮细胞,并将细胞分为正常对照组、高糖组、CIN干预组,每组6例。高糖组及CIN干预组均加入33.3 mmol/L的葡萄糖制造高糖细胞损伤模型,而CIN组同时加入0.1μmol/L CIN干预。各组细胞在培养6、24、48 h时,检测上清液中的NO水平及细胞内一氧化氮合酶(NOS)浓度;培养48 h后RT-PCR检测细胞内诱导型一氧化氮合酶(iNOS)、内皮型一氧化氮合酶(eNOS)mRNA的表达;培养48 h后Western blotting检测细胞间粘附分子-1(ICAM-1)及血管粘附分子-1(VCAM-1)蛋白的表达。结果与正常对照组相比,高糖组NO、NOS含量呈现早期升高晚期下降的趋势,而CIN组上述趋势消失;与正常组比较,高糖组内皮细胞的eNOS mRNA表达水平降低(P<0.05),iNOS mRNA、ICAM-1与VCAM-1蛋白表达明显升高(P<0.05),而CIN组eNOS mRNA表达水平较高糖组明显升高(P<0.05),iNOS、ICAM-1与VCAM-1蛋白的表达水平明显降低(P<0.05)。结论 CIN可平衡NOS的活性,调节NO的生成及ICAM-1与VCAM-1的表达,改善高糖状态下内皮细胞的功能。

Objective Nitric oxide(NO) and soluble guanylate cyclase(sGC) are both oxidized under the condition of diabetes, resulting in abnormal conduction of the nitric oxide/soluble guanylate cyclase/cyclic guanosine monophosphate(NO-sGC-cGMP) signaling pathway, which is the important pathological basis of diabetic endothelial dysfunction. Cinaciguat(CIN) is a soluble guanylate cyclase(sGC) activator that can protect endothelial cells by activating sGC. In the present study, we investigated the effect of Cinacigua on human umbilical vein endothelial cells(HUVECs) treated with high glucose in vitro. Methods The cultured HUVECs were divided into the normal control group, high glucose group and CIN group, with 6 cases in each group. High glucose group and CIN group were treated with high glucose(33.3 mmol/L), and the CIN group was also given CIN. The levels of NO and nitric oxide synthase(NOS) were measured after the intervention with 6 h, 24 h, 48 h. Forty-eight h after the intervention, mRNA expression eNOS and iNOS were examined by using RT-PCR, and the expression of ICAM-1 and VCAM-1 were examined by using Western blotting. Results Compared with normal control group, the levels of NO and NOS in the high glucose group increased at early stage and decreased at later stage(P<0.05), but CIN could balance the levels of NO and NOS(P<0.05). In the same time, the expressions of eNOS mRNA was down-regulated, In addition, ICAM-1 and VCAM-1 were up-regulated in the state of high glucose, but the level of eNOS mRNA in CIN group was higher than that in high glucose group, and the level of iNOS mRNA, ICAM-1 and VCAM-1 decreased obviously(P<0.05). Conclusion CIN can balance the activity of NOS, adjust the expression of NO, ICAM-1 and VCAM-1 of HUVECs induced by high glucose, and improve function of injured HUVECs.