The Study Of Effect Of Interleukin-10 On Renal Tubular Epithelial To Mesenchymal Transition And Relevant Mechanisms

Objective To investigate the effect of interleukin-10(IL-10) on epithelial to mesenchymal transition in human renal tubular cells and relevant mechanisms.Methods Human renal proximal tubular cell line HK-2 cell was used as target cell. These cells were divided into five groups randomly:①control group:HK-2 cells were incubated with 5.5mmol/L D-glucose.②high glucose group:HK-2 cells were incubated with 30 mmol/L D-glucose.③1ng/ml IL-10 group: HK-2 cells were incubated with 30 mmol/L D-glucose and supplemented with 1 ng/ml IL-10.④5ng/ml IL-10 group: HK-2 cells were incubated with 30 mmol/L D-glucose and supplemented with 5 ng/ml IL-10.⑤25ng/ml IL-10 group: HK-2 cells were incubated with 30 mmol/L D-glucose and supplemented with 25ng/ml IL-10. 48 hours later, inverted microscope was used to observe the morphological change, immunocytochemistry was used to measure the expression of E-cad and α-SMA, RT-PCR was used to exam the expression of PDGF mRNA and CTGF mRNA, western blot was used to check the expression of PDGF,E-cad andα-SMA,and then ELISA was used to determine the content of FN in supernatant.Results (1) Cells morphological:HK-2 cells in control group showed a clssic cobblestone morphology,in high glucose group HK-2 cells were elongated in shape, disassociating from neighboring cells, losing their cobblestone monolayer pattern and becoming fusiform,and these cells in l ng/ml IL-l0 group,5 ng/ml IL-l0 group and 25 ng/ml IL-l0 group were a cobblestone morphology in a part and becoming fusiform in another part.(2)Compared with control group, the expression of E-cad was low and the expression of PDGF, CTGF,α-SMA and FN was high in high glucose group (P<0.05),which implied high glucose can induce HK-2 cells epithelial to mesenchymal transition. (3)Compared with high glucose group, the expression of E-cad was high and the expression of PDGF, CTGF,α-SMA and FN was low in l ng/ml IL-l0 group,5 ng/ml IL-l0 group and 25 ng/ml IL-l0 group (P<0.05),which indicated that IL-10 can inhibited HK-2 cells epithelial to mesenchymal transition.Conclusion High glucose can induce HK-2 cells epithelial to mesenchymal transition and ECM production by increasing the expression of PDGF and CTGF,and then IL-10 can inhibited HK-2 cells epithelial to mesenchymal transition and ECM production by decreasing the expression of PDGF and CTGF.