高糖对肾小管上皮细胞Toll样受体4表达的影响

目的本实验旨在探讨高糖对。肾小管上皮细胞Toll样受体4（TLR4）表达的影响及其在糖尿病。肾病中的意义。方法体外培养。肾小管上皮细胞（NRK-52E），分成LG组（给予5mmol／L葡萄糖的DMEM培养）和HG组（给予25mmol／L葡萄糖的DMEM培养），不同时间收集细胞，采用细胞免疫化学、Rt—PCR、WesternBlot检测TLR4蛋白和mRNA表达情况，同时取细胞培养上清液用Elisa法检测IL-6、TNF-α水平。结果在高糖刺激6hTLR4mRNA出现表达明显增高，维持较高表达至274h，而48h后表达有所下降（P〈0．05）。HG组培养24hTLR4蛋白升高，48hTLR4蛋白表达进一步升高，与LG组比较差异有统计学意义（P〈0．05）。HG组NRK-52E上清液IL-6、TNF一仅的表达较LG组明显升高（P〈O．05），并与TLR4蛋白呈正相关（r=0．799，0.820）。结论高糖可能通过上调NRK-52E的TLR4的表达，导致TNF-α、IL-6等炎性因子表达升高，TLR4参与了糖尿病肾病的进展。

The effect of hyperglycemia on the expression of Toll-like receptor 4 in renal tubular epithelial cells

Objective This study was designed to explore the effect of hyperglycemia on the expression of Toll-like receptor 4 (TLR4 ) in renal tubular epithelial cells and its significance in diabetic nephropathy. Methods In vitro cultured renal tubular epithelial cells ( NRK-52E) were divided into LG group (cultured in 5mmol / L glucose DMEM) and HC group (cultured in 25mmol / L glucose DMEM). Cells were harvested at different time points. Immunohistochemistry, Rt-PCR, Western Blot were used to detect TLR4 protein and mRNA expression, and the levels of IL-6 and TNF-α from the cell culture supernatant were determined by EL1SA assay. Results After 6 hours, there was increased expression TLR4 mRNA in HC group, which appeared to be maintained for 24 hours and began to decrease after 48 hours ( P < 0.05). TLR4 protein expression increased in HC group after 24 hours, and increased even further after 48 hours. Compared with LG groups, the difference had statistical significance ( P <0.05). In HG group, IL-6 and TNF-α expression in the supernatant from the NRK-52E culture were significantly increased ( P < 0.05) , and the expression of IL-6 and TNF-α was positive correlated with the expression of TLR4 protein ( r =0.799,0.820). Conclusion High glucose triggers an increase in expression of TLR4 in NRK-52E cells, itself leading to an increase in expression of inflammatory factors such as TNF-α and IL-6. In this way, TLR4 participates in the progress of diabetic nephropathy.