白藜芦醇对高糖环境下人视网膜血管内皮细胞增殖的影响及分子机制

doi:10.6040/j.issn.1671-7554.0.2018.799

摘要/Abstract

摘要： 目的观察白藜芦醇(Res)对高糖环境下人视网膜血管内皮细胞(HRCECs)增殖和细胞周期的调控作用,并对其可能的分子机制进行初步探讨。方法细胞分组及处理如下:对照组(N组)加入5.5 mmol/L葡萄糖;高渗对照组(M组)加入5.5 mmol/L葡萄糖和19.5 mmol/L甘露醇;高糖对照组(H组)加入25 mmol/L葡萄糖;实验组(R1~R4组)加入25 mmol/L葡萄糖+ 25、50、100、200 μmol/L Res,分别进行培养。采用CCK-8法检测Res对HRCECs增殖的影响;流式细胞术检测Res对HRCECs细胞周期的影响;分别采用real-time PCR和Western blotting法检测PCNA、Cyclin A1、CDK2及P21^CIP1的mRNA和蛋白水平。结果 CCK-8结果显示,Res可抑制高糖诱导的HRCECs增殖活性,并且呈浓度依赖性。流式细胞术分析结果显示,与H组相比,R2组和R3组处于S期的细胞数增加(P<0.01)。 Real-time PCR和Western blotting分析结果显示,与N组相比,H组的Cyclin A1、CDK2和PCNA mRNA及蛋白的表达量增加,P21^CIP1 mRNA及蛋白的表达量下降(P<0.01);与H组相比,R2组和R3组Cyclin A1、CDK2和PCNA的mRNA及蛋白表达量降低,而P21^CIP1的mRNA及蛋白表达量增加(P<0.01)。结论 Res可通过下调Cyclin A1、CDK2和PCNA的mRNA、蛋白表达量及上调P21^CIP1 mRNA、蛋白表达量抑制高糖诱导的HRCECs的增殖活性。

关键词: 白藜芦醇, 人视网膜血管内皮细胞, 周期相关蛋白, 细胞周期

Abstract: Objective To investigate the effect of resveratrol(Res)on the proliferation and cell cycle of human retinal capillary endothelium cells(HRCECs)in high glucose environment, and to explore its underlying mechanism. Methods HRCECs were divided into 5 groups with different treatment: Control group(N group)was treated with 5.5 mmol/L glucose; hypertonic group(M group)was treated with 5.5 mmol/L glucose and 19.5 mmol/L mannitol; high glucose control group(H group)was treated with 25 mmol/L glucose; experiment groups(R1-R4 groups)were treated with 25 mmol/L glucose and 25, 50, 100, 200 μmol/L Res. CCK-8 assay and flow cytometry were used to detect the effect of Res on the proliferation and cell cycle of HRCECs, respectively. Real-time PCR and Western blotting were used to detect the mRNA and protein levels of PCNA, Cyclin A1, CDK2 and P21^CIP1, respectively. Results The proliferation 山东大学学报 (医学版)57卷3期 -崔锡铭,等.白藜芦醇对高糖环境下人视网膜血管内皮细胞增殖的影响及分子机制 \=-of HRCECs induced by high glucose was inhibited by Res in a concentration-dependent manner. Compared with H group, the cell numbers in S phase were increased in R2 and R3 groups(both P<0.01). Compared with N group, the mRNA and protein expressions of Cyclin A1, CDK2 and PCNA were increased while those of P21^CIP1 were decreased in H group(all P<0.01). Compared with H group, the expressions of Cyclin A1, CDK2 and PCNA in HRCRCs were decreased and those of P21^CIP1 were increased in R2 and R3 groups(all P<0.01). Conclusion Res can inhibit the high glucose-induced proliferation by down-regulating the expressions of Cyclin A1, CDK2 and PCNA and up-regulating the expression of P21^CIP1 in HRCECs.

Key words: Resveratrol, Human retinal capillary endothelium cells, Cycle-related proteins, Cell cycle

中图分类号:

R587.2

崔锡铭,王霜,许顺江,张睿,谢冰,崔冬生,赵占胜. 白藜芦醇对高糖环境下人视网膜血管内皮细胞增殖的影响及分子机制[J]. 山东大学学报 (医学版), 2019, 57(3): 19-24.

CUI Ximing, WANG Shuang, XU Shunjiang, ZHANG Rui, XIE Bing, CUI Dongsheng, ZHAO Zhansheng. Effect of resveratrol on the proliferation of human retinal capillary endothelial cells induced by high glucose and its possible mechanism[J]. Journal of Shandong University (Health Sciences), 2019, 57(3): 19-24.

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