摘要
目的:探讨白藜芦醇(RSV)对体外高糖刺激引起大鼠肾小球系膜细胞(RGMCs)损伤的影响及其潜在分子保护机制。方法:取处于对数生长期的RGMCs,分为甘露醇高渗透压对照组(OC)、正常对照组(NG)、高糖组(HG)以及高糖RSV处理组,分别处理24、48 h。试剂盒检测不同时间各组细胞上清液中LDH的水平;Western印迹法检测不同时间各组凋亡相关B淋巴细胞瘤-2基因(Bcl-2)、Bcl-2相关X蛋白(Bax)、半胱氨酸天冬氨酸蛋白酶(Caspase)-1、Caspase-1活性片段(p20)、Gasdermin E(GSDME)、GSDME活性片段(GSDME-N)、Caspase-3、Caspase-3活性片段(p17)、炎性相关细胞因子白细胞介素-1β(IL-1β)及其活化形式(mIL-1β)的表达;ELISA检测不同时间各组细胞培养上清IL-1β的水平。结果:LDH结果显示,RSV可降低高糖刺激48 h后细胞的死亡率(F=31.48,P<0.05)。Western印迹法结果显示,仅在RSV处理24 h后,Bax表达较HG组显著降低,同时伴随Bcl-2表达升高(F=143.1,P<0.05);RSV处理24 h后HG组p20、mIL-1β表达降低(F=26.74、25.76,均P<0.05);此外,RSV处理48 h后p17、GSDME-N的表达较HG组明显下降(F=25.84、18.49,均P<0.05)。ELISA结果显示,与HG组相比,RSV处理48 h后上清中IL-1β的水平显著降低(F=80.22,P<0.01)。上述指标在OC组及NG组间的变化均无统计学意义(均P>0.05)。结论:RSV可通过减少炎症因子的产生及抑制细胞早期凋亡、晚期焦亡过程来改善高糖引起的RGMCs损伤。
Objective:To investigate the effect of resveratrol(RSV)on the injury of rat glomerular mesangial cells(RGMCs)induced by high glucose in vitro and its potential molecular protection mechanism.Methods:RGMCs in logarithmic growth phase were divided into mannitol hypertonic control group(OC),normal control group(NG),high glucose group(HG)and high glucose with RSV treatment group,and RGMCs were cultured for 24 h and 48 h respectively.The levels of LDH in the supernatant of each group were detected by the LDH release assay kit.The protein levels of apoptosis related B-cell lymphoma-2 gene(Bcl-2),Bcl-2 associated X protein(Bax),cysteine-containing aspartate specific protease 1(Caspase-1),Caspase-1 activated fragment(p20),Gasdermin E(GSDME),Gasdermin E activated fragment(GSDME-N),Caspase-3,Caspase-3 activated fragment(p17),inflammatory related cytokines interleukin-1β(IL-1β)and mature form of IL-1β(mIL-1β)were detected by Western blotting analysis.The levels of IL-1βin the supernatant were detected by ELISA analysis.Results:LDH assay showed that RSV reduced the cell death rate after 48 h of high glucose stimulation(F=31.48,P<0.05).Western blotting analysis showed that the expression of Bax in RSV treatment group was significantly lower than that in HG group only at 24 h and the expression of Bcl-2 was increased(F=143.1,P<0.05).RSV also decreased the expression of p20 and mIL-1βin HG group at 24 h(F=26.74,25.76,all P<0.05).Moreover,RSV significantly decreased the expression of p17 and GSDME-N in HG group at 48 h(F=25.84,18.49,all P<0.05).ELISA analysis showed that IL-1βlevel in the supernatant of RGMCs with RSV treatment for 48 h was significantly reduced than that in HG group(F=80.22,P<0.01).There were no significant difference in above related indexes between OC group and NG group(all P>0.05).Conclusion:RSV can improve the damage in RGMCs induced by high glucose via reducing the production of inflammatory factors,inhibiting the early apoptotic and late pyroptosis.
作者
王琳
刘亚珊
刘瑞卿
高爱娇
唐慧欣
申艳娜
WANG Lin;LIU Ya-shan;LIU Rui-qing;GAO Ai-jiao;TANG Hui-xin;SHEN Yan-na(School of Medical Laboratory,Tianjin Medical University,Tianjin 300203,China)
出处
《天津医科大学学报》
2021年第4期338-342,共5页
Journal of Tianjin Medical University
基金
国家自然科学基金(32071263,81772252)。