摘要
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。
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 HG 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 ELISA assay. Results After 6 hours, there was increased expression TLR4 mRNA in HG group, which appeared to be maintained for 24 hours and began to decrease after 48 hours （ P 〈 0. 05）. TLR4 protein expression increased in HG 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-ot 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.
出处
《中国医师杂志》
CAS
2010年第1期63-67,共5页
Journal of Chinese Physician
