中国全科医学Issue(20)：2433-2438,6.DOI:10.3969/j.issn.1007-9572.2015.20.018
人红细胞生成素对高糖诱导肾近曲小管上皮细胞转分化过程中炎性因子的影响及其可能机制
Effect and Possible Mechanism of rhEPO in the Transdifferentiation of Human Kidney Proximal Tubular Epithelial Cells Induced by High Glucose and the Changes of Inflammatory Cytokines
摘要
Abstract
Objective To investigate the effect and possible mechanism of rhEPO in the transdifferentiation of human kidney proximal tubular epithelial cells( HK -2 cells) induced by high glucose and the changes of inflammatory cytokines. Methods By using random number table,HK-2 cells cultured in vitro were divided into several groups:blank control group ( with no irritants),high glucose induced group( with a high glucose final concentration of 30 mmol/L),mannitol control group ( with a mannitol concentration of 24. 5 mmol/L),rhEPO control group( with a rhEPO final concentration of 20 U/ml),three rhEPO intervention groups(with a rhEPO final concentration of 5,10 and 20 U/ml respectively and high glucose added),Rho kinase inhibitor(Y27632)group(in which Y27632 with a final concentration of 30 μmol/L was added and high glucose was added 30 minutes later with a final concentration of 30 mmol/L). After 24 h in virto culture,reverse transcription polymerase chain reaction(RT-PCR)was used to evaluate the mRNA levels of RhoA and ROCK. The levels of E-cadherin and α-smooth muscle actin(α-SMA) proteins were assessed by immunofluorescent test. ELISA was used to measure the levels of Fibronectin(FN),IL-6 and TNF-αproteins. Results The groups were all significantly different(P<0. 05)in the mRNA levels of RhoA and ROCK;the high glucose induced group and the 5 U/ml rhEPO intervention group were higher ( P<0. 05 ) than the blank control group in the mRNA levels of RhoA and ROCK;the 10 U/ml rhEPO intervention group and the 20 U/ml rhEPO intervention group and the Y27632 group were lower ( P <0. 05 ) than the blank control group in the mRNA levels of RhoA and ROCK;the three rhEPO intervention groups were lower ( P <0. 05 ) than the high glucose induced group in the mRNA levels of RhoA and ROCK;the Y27632 group was lower(P<0. 05)than the high glucose induced group in the mRNA levels of ROCK;the 10 U/ml rhEPO intervention group and the 20 U/ml rhEPO intervention group were lower(P<0. 05)than the 5 U/ml rhEPO intervention group in the mRNA levels of RhoA and ROCK;the 20 U/ml rhEPO intervention group was lower (P<0. 05)than the 10 U/ml rhEPO intervention group in the mRNA levels of RhoA and ROCK. The groups were significantly different(P<0. 05)in the protein levels of α -SMA,E -cadherin,FN,IL -6 and TNF -α;the high glucose induced group,the three rhEPO intervention groups and the Y27632 group were higher(P<0. 05)in the protein levels of α-SMA, FN,IL-6 and TNF-αand were lower(P<0. 05)in E-cadherin than the blank control group;the three rhEPO intervention groups and the Y27632 group were lower(P <0. 05)in the protein levels of α -SMA,FN,IL -6 and TNF -α and were higher(P<0. 05)in E-cadherin than the high glucose induced group;the 10 U/ml rhEPO intervention group and the 20 U/ml rhEPO intervention group were lower(P<0. 05)in the protein levels ofα-SMA,FN,IL-6 and TNF-αand higher(P<0. 05)in E-cadherin than the 5 U/ml rhEPO intervention group;the Y27632 group was lower(P<0. 05)in the protein levels ofα-SMA and FN and were higher(P<0. 05)in E-cadherin than the 5 U/ml rhEPO intervention group;the 20 U/ml rhEPO intervention group was lower(P<0. 05)in the protein levels of α-SMA,FN,IL-6 and TNF-αand was higher(P<0. 05) in E - cadherin than the 10 U/ml rhEPO;the Y27632 group was lower ( P <0. 05 ) than the 10 U/ml rhEPO intervention group in the protein levels ofα-SMA,E-cadherin and FN. The pearson linear correlation analysis showed that the mRNA level of RhoA was positively correlated with the mRNA level of ROCK1 in the high glucose group and the three rhEPO intervention groups(r=0. 885,0. 901,0. 886,0. 868;P<0. 05). Conclusion rhEPO could inhibit the transdifferentiation of HK-2 cells induced by high glucose to delay the fibrosis of renal. rhEPO can also reduce the production of inflammatory cytokines,the generation of inflammatory cytokines and inflammatory response, thus delaying the progression of diabetic nephropathy and the fibrosis of renal interstitium. The mechanism may be related to RhoA/ROCK signaling pathway.关键词
糖尿病肾病/红细胞生成素/细胞转分化/白细胞介素6/肿瘤坏死因子α/RhoA/ROCK信号通路Key words
Diabetic nephropathies/Erythropoietin/Cell transdifferentiation/Interleukin - 6/Tumor necrosis factor-alpha/RhoA/ROCK signaling pathway分类
医药卫生引用本文复制引用
陈艳霞,杨丽萍,吴险峰,秦晓华,黄翀,房向东,涂卫平..人红细胞生成素对高糖诱导肾近曲小管上皮细胞转分化过程中炎性因子的影响及其可能机制[J].中国全科医学,2015,(20):2433-2438,6.基金项目
江西省自然科学基金资助项目 （）
