The Study Of The Inhibiting Effect Of Baical Bai On Rat Glomerular Mesangial Cell Proliferation Induced In High Glucose By Down-regulation Of NF-κB Expression

ObjectiveTo study the inhibiting effect of Baical Bai on rat glomerular mesangial cell(GMC) proliferation induced by high glucose(HG)and the effect of Nuclear Factor Kappa B(NF-κB) expression. This research enrichs complex cytokine network of Diabetic Nephropathy kidney, reveals the pharmaco-mechanism of Baicalin efficacy on control DN, and to provide experimental basis for curative effect on DN with Chinese medicine.MethodsThis experiment divided rat GMC culture in vitro into six groups:(1)Low glucose group(LG5.5mmol/L),(2) High glucose group(HG25mmol/L),(3) Baicalin Bai in the low dose group (HG+baicalin25μmol/L),(4) Baicalin Bai in the middle dose group (HG+baicalin50μmol/L),(5) Baicalin Bai in the high dose group (HG+baicalin100μmol/L),(6)Losartan group(HG+Losartan potassium10μmol/L). Each group set six parallel holes, training24h,48h and72h. At the end of each time piont, used MTT colorimetric assay to detect the GMC growth situation. According to its results, choose the best action time. And then through the immunohistochemical method to detect FN protein expression. Collect cells and extract the nucleoprotein, by the Western blot test method to detect the Nuclear Factor Kappa B(NF-κB) expression.Results(1)The absorbance of HG group is higher than LG group in24h,48h and72h, in addition the absorbance increased significantly with the cultivate time extending(P<0.05); The absorbance of Baicalin Bai in the low dose group, Baicalin Bai in the middle dose group, Baicalin Bai in the high dose group, Losartan group were lower than HG group in24h, while higher than HG group in48h and72h (P<0.05). The inhibition ratio of Baicalin Bai and Losartan were both negative in24h, while the positive value in48h and72h. so, the inhibition rate of Baicalin Bai was increased with the increased concentration and the best inhibit effect is in72h.(2)By combining the positive reaction with shading degree to evaluate immunohistochemical results, which indicated that the positive rate of FN expression in HG group was much more than other groups, and the shading degree was stronger than other group too.After Baicalin Bai intervention, both of positive expression and shading degree were weaken. The FN expression in Losartan group was weak positive, but wide range of it.(3) Western blot results showed that NF-κB expression in HG group was higher than Low glucose group. And with the Baicalin Bai intervention, its expression was reduced. With the concentration increase, expression was reduced gradually. There was no obvious difference of NF-κB expression betwent the HG group and Losartan group.ConclusionProliferation of rat GMC culture in vitro was promoted by high glucose, and Baicalin Bai can restrain this phenomenon, which was relevant to Baicalin Bai Reduce expression of NF-κB.