The Role Of FOXM1 In High Glucose-induced Collagen Synthesis In MRC-5 Cells

Objective:To investigate the role of FOXM1 in collagen synthesis of MRC-5 cells induced by High glucose(HG).Methods:MRC-5 cells were induced by high glucose to establish diabetic pulmonary fibrosis model.After the cells were treated with different glucose concentration and time,the cell proliferation was measured by CCK8 to determine the best condition for high glucose modeling;Western blot was used to detect the expressions of collagen synthesis and TGF-?/Smads pathway factors,and the m RNA expressions of FOXM1,collagen synthesis and TGF-?/Smads pathway factors were deceted by q PCR assay.Results:(1)the cells were treated with different time and concentration of glucose.The results of CCK8 showed that the proliferation activity of cells decreased with time or concentration of glucose increasing.When the cells were cultured for 24 h,the cells proliferation began to decrease obviously,and the cells proliferation activity continued to decrease at 48h and 72 h with time prolonging,(p<0.05).There was no significant difference between HM and CON(p>0.05).When the concentration of glucose was 30mmol L·^-1,the cells proliferation began to decrease significantly and with the concentration of glucose increasing,the proliferation rate went on lower than that of the control group(p<0.05).Thus,the best conditions for high glucose modeling were 30 mmol L·^-1glucose and 24 h.(2)The cells were divided into normal control group(CON),30mmol·L^-1hypertonic control group(HM)and 30mmol·L^-1high glucose group(HG).The results showed that there were no significant changes in protein expression and m RNA level between HM and CON.The levels of Col III,Col I,Fn and TGF-?/Smads pathway factors in high glucose group were significantly higher than those in control group,while the levels of MMP9protein and m RNA were significantly lower than those in control group(p<0.05).(3)The cells were divided into normal control group(CON),30mmol·L^-1hypertonic control group(HM),30mmol·L^-1high glucose group(HG),30mmol·L^-1high glucose+1?mol thiostrepton group(HG+TST),and the protein expressions and m RNA levels were detected after 24 h.The results showed that there were no significant changes in protein expression and m RNA level between HM and CON.The levels of Col III,Col I,Fn,?-SMA,TGF-?1 and other factors in high glucose group were significantly higher than those in control group,while the levels of MMP9 protein and m RNA were significantly lower than those in control group(p<0.05).After TST treatment,the protein and m RNA expression level of Fn,FOXM1,Col III,Col I,?-SMA and TGF-?1 were significantly decreased,while the protein expression and m RNA level of MMP9 were significantly increased(p<0.05).Conclusion:FOXM1 is a factor related to the increase of fibrosis and collagen synthesis induced by high glucose.The specific inhibitor of FOXM1,TST,can reverse the increase of collagen synthesis induced by high glucose.The mechanism may be related to the TGF-?/Smads pathway.