Effect Of Advanced Glycation End Products On The Expression Of Fibrosis Associated Factors In Cultured Human Renal Mesangial Cells

Objective In this study,the effects of AGEs on transforming growth factorβ1(TGF-β1),connective tissue growth factor(CTGF)and fibronectin(Fn)mRNA expression in cultured human renal mesangial cell(sHRMC)were examined,and then RAGE was blocked by neutralizing antibodies to RAGE that was used to explore the possible mechanisms that how AGE-BSA induced the expression of fibrosis factor.Methods Human renal mesangial cells were incubated with medium containing different doses of AGE-BSA or BSA(50,100,200,400mg/L) for 48 hours,or with AGE-BSA 200 mg/L for different times (0,12,24,48,72h).Cell proliferation of HRMC cells was measured by MTT.Western blotting assay was used to estimate the protein level of RAGE.The mRNA expression level of TGF-β1,CTGF and FN were analyzed by semi-quantiative reverse transcription-polymerase chain reaction(RT-PCR).ELASA was used to estimate the protein level of TGF-β1.Results (1)Following the treatment of different doses or times of AGE-BSA or BSA,the cell proliferation rate was up-regulation as the treatment concentration increased and the culture time extended.The effects were dose and time-dependent(P<0.05).(2)Compared with the blank or BSA group,after the treatment of AGE-BSA,the protein level of RAGE was incresed,AGE-BSA significantly increased the expressions of TGF-β1,CTGF,FN mRNA and RAGE in a time- and dose-dependent manner as well(P<0.05).(3)The effect of AGE-BSA-induced up-regulated of TGF-β1,CTGF,FN mRNA was significantly blocked by treatment with neutralizing antibodies to RAGE,while the espression of TGF-β1,CTGF,FN mRNA stood nearly unchanged after cultured with huamn IgG.Conclusion (1)AGEs could significantly stimulated the proliferation and expression of fibrosis associated factors in cultured human renal mesangial cells.(2)AGEs could obviously increase expression of TGF-β1,CTGF,FN mRNA in human renal mesangial cells through RAGE.