脂联素对高糖环境下人肾小球系膜细胞增殖及氧化应激水平的影响

Effects of adiponectin on proliferation and oxidative stress level of HMCs cultured with D-glucose of high concentration

目的探讨脂联素(Adiponectin,ADPN)对高糖环境下人肾小球系膜细胞(HMCs,humanMesangial Cells)增殖及氧化应激水平的影响,揭示脂联素在糖尿病肾病中的保护作用。方法①体外培养HMCs:①将HMCs随机分为正常对照组(5.5mmol/L葡萄糖)、高糖组(30.0mmol/L葡萄糖)、脂联素组(30.0mmol/L葡萄糖+2.5μg/ml脂联素),分别培养24、48、72h后,运用四甲基偶氮唑蓝(MTT)法测定3组不同时间点HMCs的增殖情况。②将HMCs随机分为上述3组细胞,分别培养24、48、72h后运用硫代巴比妥酸法(TBA法)测定细胞上清液中MDA及SOD的含量。③再将HMCs随机分为上述3组细胞,分别培养24、48、72h,运用实时荧光定量PCR(RT-PCR)测定HO-1mRNA的表达,观察细胞氧化应激指标变化。结果①24h高糖组与正常对照组比较细胞增殖无统计学差异(P>0.05),48、72h两组比较,高糖组细胞增殖明显受抑制(P<0.05)。加入脂联素促进细胞增殖,与高糖组比较差异有统计学意义(P<0.05);②24、48、72h高糖组MDA含量均升高,SOD含量及HO-1mRNA的表达均下降,与正常组比较差异有统计学意义(P<0.05)。加入脂联素后,观察上述氧化应激指标均得到明显改善,差异有统计学意义(P<0.05)。结论长期高糖刺激抑制人肾小球系膜细胞增殖,外源性加入脂联素可促进细胞增殖。高糖组均引起氧化应激指标升高,加入脂联素可降低系膜细胞氧化应激水平,提示其对细胞起保护作用。

Objective To investigate the effects of adiponectin（ADPN） on cell proliferation and the oxidative stress level of human mesangial cell cultured with high concentration of D-glucose and to illuminate the potential protection of ADPN on the kidney.Methods The human mesangial cells（HMCs） culture in vitro ①The human mesangial cells were cultured with 5.5 mmol/L glucose（control group）,30.0 mmol/L glucose（high glucose group）,30.0 mmol/L glucose and concentration of 2.5 μg/ml adiponectin（ADPN group）in random.After culture for 24,48,72 h,the proliferation activity of human mesangial cell was observed by MTT.②After HMCS was cultured with upper three different kinds of environments for 24,48,72 h.the content of MDA and SOD of the cell supernatant was measured by Thibabituric acid（TBA）.③After HMCS was cultured with upper three different kinds of environments for 24,48,72 h.the expression of HO-1 mRNA was observed by RT-PCR.Results Compared with normal group,HMCs proliferation activity has no difference in high glucose group in 24 h（P0.05）,and that was inhibited in 48 h,72 h（P0.05）.but the proliferation of HMCS was increased after adiponectin being added（P0.05）.Meanwhile,the content of SOD and the level of mRNA of HO-1 decreased and the content of MDA increased in the high glucose group in the 24,48,72 h compared with normal group（P0.05）.After adding adiponectin,the content of SOD MDA and the level of mRNA of HO-1 had being ameliorated and there has statistical difference（P0.05）.Conclusion Long-term high glucose stimulating could inhibit cell proliferation of human mesangial cells,but adiponectin could promote the proliferation of the HCM and protect them in D-glucose of high concentration.The high glucose could enhance the oxidative stress level,but adiponectin could ameliorate it.So adiponectin could protect the mesangial cell in high density glucose.