Kidney Security I On Rat Mesangial Cell Proliferation And Tgf-beta <sub> 1 </ Sub> Of Hgf Expression In Experimental Research

Objective:To observe the cell proliferation and expression of Transforming growth factor-β1 (TGF-β1) and Hepatocyte growth factor (HGF ) in rat glomerular mesangial cells(GMCs) cultured in vitro. And to study the mechanism of Shen' an I in inhibiting the renal fibrosis. Methods:1. The GMCs stimulated with Lipopolysaccharide(LPS) were cultured in vitro.2. The GMCs were divided into three groups : the Normal group (cultured with common culturing fluid), LPS group (cultured with culturing fluid including LPS ), and Shen' an group (cultured with culturing fluid including Shen' an I ). After culturing 24 and 48 hours, the morphological variation characteristics of rat's GMC can be seen through the microscope.3. The GMCs's activity with different concentrations of Shen' an I were checked with Trypan-blue tinctured method. And we can observe the Shen' an I 's poison to GMC and choose the proper concentration for experiment.4. There were five groups in our experiment : the GMC group, LPS group, Low-dose Shen' an group, Middle-dose Shen' an group and High-dose Shen' an group. After adding different serum respectively, the GMC proliferation level was detected by the MTT ( 3-(4,5-Dimethylthiazol-2-yl)2,5-diphenyltetrazo-liumbromid ) spectrometric assay method in corporation at the 24th, 48th and 72th hour.5. The GMCs were divided into three groups : the GMC group, LPS group and Shen' an group. After adding different serum respectively, the expression of TGF-Bi and HGF were checked by immunohistochemistry.Results:1. The rat's GMCs were living well in vitro.2. The morphological characteristics and living statue of rat's GMCs were in accord with related issues.3. The activity of GMCs among 25%,12.5% and 6.25% concentration groups after culturing 24, 48 and 72 hours were all above 90%. And have no obvious difference compared with the normal group(P>0.05).4. The OD results of MTT assay show that : in the LPS group, the GMC's proliferation level was obviously higher than the GMC group(P<0.05 or 0.01); otherwise , in the Shen' an group, the GMC's proliferation level was obviously lower than the LPS group (P<0.05 or 0.01).5. The results of immunohistochemistry show that : (1) There are positive expression in all samples differently; (2) The TGF-Bi expression of GMCs in the LPS group was obviously higher than that in GMC group (P<0.05).Otherwise, the HGF expression had no remarkable difference (P>0.05); (3) The HGF expression of GMCs in the Shen' an group was obviously higher than that in GMC group (P<0.01).At the same time, the TGF-Bi expression had no obvious difference (P>0.05); (4) The TGF-Bi expression of GMCs in the Shen' an group was obviously lower than that in LPS group (P<0.01).Conversly, the HGF expression was higher (P<0.01). Conclusion:The expeiment shows that rat's GMC is the important target cell for action of Shen' an I which can obviously inhibit the proliferation of GMCs stimulated with LPS, down-regulate the expression of TGF-Bi , and up-regulate the expression of HGF without no poison. And this may be the mechanism of its effects of anti-renal fibrosis.