| Background and purpose At present,kidney disease is related to glomerular podocyte damage,inflammation and autoimmunity.In recent years,it has been proposed that inflammatory factors play different roles in many diseases.Among them,the role of IL-27 in many clinical inflammatory diseases is deepened with the increase of cases.IL-27 can not only enhance the proliferation and differentiation of Th1 cells,but also promote inflammatory diseases to some extent,and inhibit the proliferation and differentiation of Th1 and Th2 cells,so it also has anti-inflammatory effect in inflammatory diseases,IL-27 has anti-inflammatory and pro-inflammatory effects,so IL-27 can play an important role in anti infection,anti-tumor,immune and other related diseases.In this study,a mouse model of podocyte injury induced by puromycin aminonucleoside(Pan)was constructed in vitro,and tacrolimus(FK506)was given to intervene.The changes of IL-27 concentration were observed by ELISA,and the cultured podocytes were collected for real-time fluorescent quantitative PCR(RT-PCR)to detect the changes of IL-27 m RNA expression The expression of IL-27 was detected.To investigate the effect of IL-27 on glomerular podocyte injury and FK506.Methods Mouse glomerular podocytes were cultured in vitro,and the control group,PAN group and PAN+FK506 group were set up.The control group was treated with PRMI1640 medium containing 0.02% DMSO;the PAN group was treated with PRMI 1640 medium supplemented with PAN(50 mg/L);the PAN+FK506 group was treated with PRMI 1640 medium supplemented with PAN and FK506(5 mg/L).After 8h,24 h and 48 h of treatment,the morphology of podocytes in each group was observed by inverted light microscope and recorded;The change of IL-27 concentration was observed by ELISA,Real-time PCR was used to detect the m RNA expression of IL-27;Western Blot was used to detect the protein expression of IL-27;Results(1)Podocyte morphology: In the control group,the podocytes were observed under microscope for 8h,24 h and 48 h respectively.The podocytes in the control group were plump,with obvious nucleus,long podocyte protrusion and close intercellular connection.Compared with the control group,in Pan Group,the volume of podocyte decreased at 8 hours,the volume of podocyte decreased at 24 hours,the volume of podocyte decreased and the volume of podocyte increased,the volume of podocyte and podocyte changed the most at 48 hours,and the space between cells was the largest.Compared with Pan Group,the podocyte body of FK506 group was significantly larger than that of Pan Group at 8h,24 h and 48 h,the difference was statistically significant,there was no significant difference between FK506 group and the control group,the podocyte process was obvious,the connection between cells was closer,the podocyte process was obvious,and the adjacent cells were closer.(2)The change of IL-27 concentration: the results of ELISA showed that the concentration of IL-27 in the control group had no significant change at 8h,24 h and 48h(P > 0.05);the concentration of IL-27 in the pan group began to increase at 8h,and increased more significantly at 24 h and 48h(P < 0.05)Compared with the control group,the concentration of IL-27 in Pan group was higher at 8h,24 h and 48h;there was no significant change in the concentration of IL-27 in tacrolimus group at 8h,with a slight increase at 24 h and 48h;compared with Pan Group,the concentration of IL-27 in tacrolimus group was lower at 8h,24 h and 48h;compared with the control group,the concentration of IL-27 in tacrolimus group was no significant difference at 8h(P > 0.05),but higher at 24 h and 48h(P < 0.05).(3)RT-PCR: the expression of il-27 mrna in the normal podocytes of the control group did not change significantly at 8h,24 h and 48h(P > 0.05);the expression of il-27 mrna in the pan group began to increase at 8h,especially at 24 h and 48 h,with the highest expression at 48 h.Compared with the control group,the expression of IL-27 m RNA in Pan group was higher at 8h,24 h and 48 h than that in the control group(P < 0.05);there was no significant change in the expression of IL-27 m RNA in tacrolimus group at 8h,but slightly increased at 24 h and 48 h.Compared with Pan Group,the expression of IL-27 m RNA in FK506 group was lower than Pan Group at all time points(P < 0.05).(4)Western blot: The expression of IL-27 protein in control normal foot cells was lower at 8 h,24 h and 48 h.The expression of IL-27 protein was significantly increased at8 h,24 h and 48 h in the PAN group compared with the control group(P < 0.05).the expression of IL-27 protein was not significantly different at 8 h in the FK506 group compared with the PAN group(P < 0.05)and decreased after 24 h(P < 0.01)and was more significant after 48 h(P < 0.01).Conclusions IL-27 can regulate the proliferation and transformation of T cells,enhance the proliferation and differentiation of T cells to Th1 and increase the secretion of interferon γ(Interferon γ,INF-γ),tumor necrosis factor-α(TNF-α)and IL-12,and also reduce the proliferation and transformation of other types of cells such as Th2 and Th17,promote inflammation and cause cell damage.IL-27 is involved in the pathogenesis of kidney disease,the application of FK506 can reduce the expression of IL-27 to a certain extent and achieve the reduction of glomerular foot cell damage,providing a certain experimental basis for the clinical application of FK506 to treat kidney disease. |
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