Effect Of HB-EGF On Proliferation And TGF-β₁ Expression Of Cultured Rat Glomerular Mesangial Cells

Objective To investigate the effect of HB-EGF on proliferation and TGF-β₁ expressionof cultured rat glomerular mesangial cells.Methods Primary cultured glomerular mesangial cells were prepared from month 1-2SD rats. Mesangial cells were identified by phase contrast microscope,scanning electronmicroscope and immunohistochemistry. The primary cultured SD rat glomerularmesangial cells were used as cell models.MTT colorimetric assay assessed theproliferation of cells. The protein release of TGF-β₁ in mesangial cells was measured byELISA and the expression of TGF-β₁mRNA was measured by RT-PCR.Results 1. 10ng/ml HB-EGF could not stimulate the proliferation of mesangial cellsï¹™P>0.05﹚, but it in the concentration of 100ng/ml and 1000ng/ml could obviouslystimulate the proliferation of cellsï¹™P<0.01﹚. The most remarkable proliferation effectwas observed by 100ng/mlï¹™P<0.01﹚. 2. HB-EGF in the concentration of 10ng/ml,100ng/ml and 1000ng/ml could stimulate the protein release of TGF-β₁ in mesangial cellsï¹™P<0.01﹚, and the peak was in the 100ng/mlï¹™P<0.05﹚. 3. mesangial cells werestimulated respectively by 100ng/ml HB-EGF for 24h,36h and 48h. The TGF-β₁ proteinrelease and TGF-β₁mRNA expression were upregulatedï¹™P<0.01﹚, and the upregulationin 36h and 48h groups was more obvious than it in 24h groupï¹™P<0.01﹚.Conclusion HB-EGF can not stimulate the proliferation of mesangial cells in lowerconcentration. But with the concentration increased, the effect of proliferation is enhancedin the range of concentration from 100ng/ml to 1000ng/ml, and it is most remarkable in100ng/ml. HB-EGF in the range of concentration from 10ng/ml to 1000ng/ml stimulatesthe protein release of TGF-β₁ in mesangial cells. 100ng/ml HB-EGF can upregulate theprotein release and TGF-β₁mRNA expression in the range of time from 24h to 48h. Bythese founds HB-EGF may have its effect in a dose-and time-dependent manner.