Effect Of Midkine In Glomerular Mesangial Cells Cultivated Under High Glucose And Intervention Of Tangshen Particle

Objective Observe the midkine in rat glomerular mesangial cells cultivated under high glucose and the changes of gene expression and activation of signaling pathway in Protein Kinase C.and its effect in the occurrence and development of diabetic nephropathy elucidated at cellular level;Study on the effect of Tangshen Particle drug serum for the damage model of glomerular mesangial cells induced by high glucose.To explore the midkine for glomerular mesangial cells induced by the high glucose which was used angiotensin converting enzyme inhibitor Benazapril as control,provided for the clinical application of experimental basis.Methods Experiments using rat glomerular mesangial cells in culture and in vitro serum pharmacological method,normal control group,model groups,the Tangshen particle Group(low,medium and high doses),Benazapril Group(control group of WM).Drugs on rat glomerular mesangial cells cultured in high glucose,the rat glomerular mesangial cells proliferative degree with in 24,48 and 72 hours were detected by MTT colorimetry.The expressions of MK、TGF-βi、PKC mRNA were detected by RT-PCR,Western Blot,respectively,and the content of MK、TGF-β1、PKC in the supernatant of cultured GMCs were detected by ELISA.Results In vitro cultured glomerular mesangial cells were obviously proliferated under the stimulation of high glucose(30.0mmol/L).Both the serum of Tangshen Particle and Benazapril have significant inhibition on proliferation of rat glomerular mesangial cells,with the former better than the latter,and certain time and dose dependence has been detected.Compared with normal control group,the content of secretion of MK、TGF-βi、PKC in the supernatant of cultured GMCs was higher in HG(p<0.001),and the expression of MK、TGF-β1、PKC was upregulated in HG group(p<0.001).Compared with HG group,HG plus Benazepril,and high,middle,low dose of Tangshen Particle significantly decreased MK、TGF-β1、PKC secretion in the supernatant of cultured GMCs(p<0.05),and decreased the expressions of MK、TGF-β1、PKC,Tangshen Particle groups were more significant(p<0.05),a certain dose dependent and time dependent was shown.Conclusions high glucose medium can promote GMCs proliferation and Tangshen Particle can inhibit GMC proliferation promoted by high glucose,and certain time and dose dependence has been detected.GMCs can increase secretion and expressions of MK、TGF-β1、PKC induced by high glucose,Tangshen Particle can reduce the secretion of MK、TGF-β1、PKC,and down regulate the expressions of MK、TGF-β1、PKC.