核转录因子κB抑制剂PDTC对高糖培养大鼠肾成纤维细胞VEGF和PEDF表达的影响

Effects of NF-κB Inhibitor PDTC on the Expression of VEGF and PEDF in Rat Kidney Fibroblasts Cultured by High Glucose

目的:探讨核转录因子κB(NF-κB)抑制剂吡咯烷二硫氨基甲酸(PDTC)对高糖培养大鼠肾成纤维细胞(NRK)血管内皮生长因子(VEGF)及色素上皮衍生因子(PEDF)表达的影响。方法:NRK按培养条件分为常糖组:5.6mmol/L葡萄糖;高糖A组:15mmol/L葡萄糖;高糖B组:30mmol/L葡萄糖;PDTC对照组:5.6mmol/L葡萄糖+5.0μmol/LPDTC;PDTCA组:15mmol/L葡萄糖+10μmol/LPDTC;PDTCB组:30mmol/L葡萄糖+20μmmol/LPT-DC。采用半定量RT-PCR方法及ELISA测定各组培养24h、48h后NRK的VEGF及PEDFmRNA及蛋白的表达。结果:与常糖组相比,高糖各组NRK的VEGF蛋白含量及mRNA表达明显升高(P<0.01,P<0.05);与高糖B组相比,PDTC干预各组NRK的VEGF蛋白含量及mRNA表达呈下降趋势(P<0.01,P<0.05)。与常糖组相比,高糖各组NRK的PEDF蛋白含量及mRNA表达明显下降(P<0.01,P<0.05);与高糖B组相比,PDTCB组NRK的PEDF蛋白含量及mRNA表达明显升高(P<0.01)。结论:PDTC能明显抑制高糖培养大鼠NRK的VEGF表达并明显上调PEDF的表达,间接提示NF-κB可能通过参与VEGF和PEDF的表达失衡导致糖尿病肾病(DN)的发生。

Objective:To explore the effect of pyrrolidine dithiocarbamate(PDTC)on the expression of vascular endothelial growth factor(VEGF)and pigment epithelium derived factor(PEDF)in rat kidney fibroblasts(NRK)cultured by different high glucose condition.Method:NRK cells were cultured in vitro,and then divided into six groups:normal glucose group:5.6mmol/L D-glucose;high glucose group A:15mmol/L D-glucose;high glucose group B:30mmol/L D-glucose;PDTC group:5.6mmol/L D-glucose +5μmol/L PDTC;PDTC group A:15mmol/L D-glucose +10μmol/L PDTC;PDTC group B:30mmol/L D-glucose +20μmol/L PDTC.The protein levels of VEGF and PEDF were measured by ELISA,and the expression of VEGF and PEDF mRNA were measured by RT-PCR.Results:Compared with normal glucose group,the protein level and mRNA of VEGF were remarkably increased when the NRK cells cultured by high glucose condition(P<0.01,P<0.05).Compared with high glucose group B,the protein level and mRNA of VEGF were remarkably decreased when the NRK cells cultured by PDTC(P<0.01).Compared with normal glucose group,the protein level and mRNA of PEDF were remarkably decreased when the NRK cells cultured by high glucose condition(P<0.01,P<0.05).Compared with high glucose group B,the protein level and mRNA of PEDF were remarkably increased when the NRK cells cultured by PDTC(P<0.01).Conclusion:PDTC induced the imbalance of expression between VEGF and PEDF in NRK cells cultured by different high glucose condition.NF-κB may play a potential role in the progression of diabetic nephropathy.