抗衰老Klotho蛋白对高糖作用下血管内皮细胞的保护作用

发布时间：2018-04-21 01:07

本文选题：高糖 + Klotho蛋白　；参考：《中国病理生理杂志》2017年01期

【摘要】：目的:研究抗衰老Klotho蛋白对高糖作用下血管内皮细胞的保护作用及其作用机制。方法:体外培养人脐静脉血管内皮细胞(HUVECs),设置PBS对照组、5.5 mmol/L葡萄糖组、33.3 mmol/L葡萄糖组、0.1μmol/L Klotho+33.3 mmol/L葡萄糖组、1μmol/L Klotho+33.3 mmol/L葡萄糖组和10μmol/L Klotho+33.3 mmol/L葡萄糖组。使用MTT法检测各组细胞活力;同时检测各组细胞培养上清中丙二醛(MDA)的含量以及乳酸脱氢酶(LDH)、超氧化物歧化酶(SOD)和还原型谷胱甘肽(GSH)的活性;流式细胞术检测各组细胞中活性氧(ROS)含量变化;ELISA检测各组细胞培养液中一氧化氮(NO)、内皮素1(ET-1)和细胞间黏附分子1(ICAM-1)的浓度变化;Western blot法检测各组细胞中核因子κB(NF-κB)蛋白的表达。结果:与PBS对照组相比,33.3 mmol/L葡萄糖能够显著降低HUVECs的细胞活力,增加细胞中ROS的含量,增加细胞培养上清中LDH活性和MDA含量,降低SOD和GSH的活性,同时降低NO分泌,诱导ET-1、ICAM-1分泌及细胞中NF-κB蛋白的表达(P0.05)。不同浓度Klotho蛋白和33.3 mmol/L高糖同时作用HUVECs时,细胞活力逐渐上升,ROS和MDA含量以及LDH活性逐渐下降,SOD和GSH活性则逐渐上升,同时NO分泌增加,ET-1、ICAM-1分泌及NF-κB蛋白表达显著下降(P0.05)。结论:抗衰老Klotho蛋白能够提升高糖作用下HUVECs的细胞活力,减少高糖诱导的ROS生成及氧化损伤,恢复HUVECs的正常分泌功能,并通过减少NF-κB蛋白表达发挥抗损伤作用。
[Abstract]:Aim: to study the protective effect of anti-aging Klotho protein on vascular endothelial cells under high glucose and its mechanism. Methods: human umbilical vein endothelial cells (HUVECs) were cultured in vitro. The PBS control group was divided into 5. 5 mmol/L glucose group and 10 渭 mol/L Klotho 33.3 mmol/L group. The mmol/L glucose group was 33.3 渭 mol/L Klotho 33.3 mmol/L glucose group, 1 渭 mol/L Klotho 33.3 mmol/L glucose group and 10 渭 mol/L Klotho 33.3 mmol/L glucose group. The activities of malondialdehyde (MDA), lactate dehydrogenase (LDH), superoxide dismutase (SOD) and reduced glutathione (GSH) were detected by MTT assay. The concentration of nitric oxide (no), endothelin 1 (et 1) and intercellular adhesion molecule 1 (ICAM 1) in culture medium of each group were detected by flow cytometry. The expression of nuclear factor- 魏 B (NF- 魏 B) NF- 魏 B protein was detected by Western blot. Results: compared with PBS control group, 33.3 mmol/L glucose could significantly decrease the cell viability of HUVECs, increase the content of ROS, increase the activity of LDH and MDA in the supernatant of cell culture, decrease the activities of SOD and GSH, and decrease the secretion of no. The secretion of ICAM-1 and the expression of NF- 魏 B protein in ET-1 cells were induced by P0.05. At different concentrations of Klotho protein and 33.3 mmol/L with high glucose, the activity of Ros and MDA and the activity of LDH decreased gradually, while the secretion of no increased the secretion of ICAM-1 and the expression of NF- 魏 B protein decreased significantly (P0. 05). Conclusion: Anti-aging Klotho protein can enhance the cell viability of HUVECs induced by high glucose, reduce the ROS production and oxidative damage induced by high glucose, restore the normal secretory function of HUVECs, and play an anti-injury role by reducing the expression of NF- 魏 B protein.
【作者单位】：三峡大学仁和医院ICU科;三峡大学仁和医院心血管内科;
【基金】：湖北省教育厅自然科学研究计划重点项目(No.D20091308)
【分类号】：R587.2;R54

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