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目的:研究可溶性鸟苷酸环化酶激活剂Cinaciguat(以下简称CIN)对高糖环境下人脐静脉内皮细胞(HUVECs)的保护作用。方法:以33.3 mmol/L的葡萄糖作用于HUVECs细胞复制氧化应激损伤模型。将HUVECs细胞分为正常组、高糖组和高糖+CIN 0.01、0.1、1、10 μmol/L组,采用MTT法检测细胞活力,计算细胞存活率;另将HUVECs细胞分为正常组、正常+CIN 1 μmol/L组、高糖组、高糖+CIN 1 μmol/L组,采用硫代巴比妥酯法、黄嘌呤氧化酶法检测细胞中丙二醛(MDA)含量和超氧化物歧化酶(SOD)活性,采用实时荧光定量聚合酶链反应法检测细胞中细胞间黏附分子1(ICAM-1)、血管细胞黏附分子1(VCAM-1) mRNA的表达。结果:与正常组比较,高糖组细胞存活率降低(P<0.05);与高糖组比较,高糖+CIN 0.01、0.1、1、10 μmol/L组细胞存活率升高(P<0.05),且呈浓度依赖性,但高糖+CIN 1 μmol/L组与高糖+CIN 10 μmol/L组间细胞存活率差异无统计学意义(P>0.05)。与正常组比较,高糖组细胞中MDA含量增加、SOD活性降低、ICAM-1和VCAM-1 mRNA表达增强(P<0.05),但正常+CIN 1 μmol/L组细胞上述指标无明显变化(P>0.05);与高糖组比较,高糖组+CIN 1 μmol/L组细胞中MDA含量降低、SOD活性增强、ICAM-1和VCAM-1 mRNA表达减弱(P<0.05)。结论:CIN对体外高糖诱导的HUVECs细胞损伤具有保护作用,其机制可能与抑制氧化应激和炎症反应有关。
OBJECTIVE: To study the protective effects of soluble ornithine cyclase activator cinaciguat (called CIN for short) on human umbilical vein endothelial cells (HUVECs) under the condition of high glucose. METHODS: HUVECs were treated with glucose 33.3 mmol/L to induce oxidative stress injury model. HUVECs were divided into normal group, high glucose group and high glucose+CIN 0.01, 0.1, 1 and 10 μmol/L groups. The cell viability was detected by MTT assay, and the survival rate of cells was callulated. Besides, HUVECs were divided into normal group, normal+CIN 1 μmol/L group, high glucose group, high glucose+CIN 1 μmol/L group. MDA content and SOD activity were determined by TBA method and xanthine oxidase method; mRNA expression of ICAM-1 and VCAM-1 were determined by RT-PCR. RESULTS: Compared with normal group, survival rate decreased in high glucose group (P<0.05); compared with high glucose group, that increased in high glucose+CIN 0.01, 0.1, 1 and 10 μmol/L groups, in concentration-dependent manner; there was no statistical significance in survival rate between high glucose+CIN 1 μmol/L group and high glucose+CIN 10 μmol/L group (P>0.05). Compared with normal group, MDA content and mRNA expression of ICAM-1 and VCAM-1 increased in high glucose group, while SOD activity decreased (P<0.05); above indicators had no significant change in normal+CIN 1 μmol/L group (P>0.05). Compared with high glucose group, MDA content and mRNA expression of ICAM-1 and VCAM-1 decreased in high glucose+CIN 1 μmol/L group, while SOD activity increased (P<0.05) . CONCLUSIONS: CIN can protect high glucose-induced HUVECs injury in vitro. The mechanism may be associated with inhibition of oxidative stress and inflammatory response.
Cinaciguat高糖人脐静脉内皮细胞氧化应激炎症反应
CinaciguatHigh glucoseHuman umbilical vein endothelial cellsOxidative stressInflammatory response
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