Combining Astragalus Injection Of Puerarin Injection On Diabetic Nephropathy Bmp-7 Pathway

Diabetic nephropathy (DN) is one of the important diabetic microvascular complications. The chief pathological characteristic is kidney glomerular hypertrophy at beginning, the basement membrane of kidney glomeruli and tubules are thickening, mesangial matrix accumulates progressively. DN can develop glomerular and interstitial fibrosis at the end stage. These changes lead to renal failure at last. As the change of the lifestyle, the incidence of ESRD, coincident with the increasing number of diabetes has raised. The pathogenesis of diabetic is still not fully understood, and there is not ideal therapy, so recent research focuses on exploreing the cause, pathogenesis and effective treatments for diabetic nephropathy. Chinese medicine has been proved by clinic that it is an effective method to treat DN; it may play a multi-target role in treating DN, so study the mechanism of TCM on treating DN is very necessary. Puerarin injection and Astragalus mongholicus inject ion has being used to treat DM and DN in clinic, they can reduce hyperglycemia, hyperlipemia, and protect renal function. We combine Puerarin injection and Astragalus mongholicus injection to treat DN, and investigate the mechanism of them to treat DN and protect renal function.ObjectiveInvestigate the mechanism of Puerarin injection combines Astragalus mongholicus injection to treat DN. Explore the specific mechanism of Chinese medicine which can reinforce qi and promote blood circulation in loss of kidney toxicity network.Methods1. KKAy mice were randomly divided into two groups:model and Puerarin injection combine Astragalus mongholicus injection group. C57BL/J mice were used to be control. Puerarin inject ion combines Astragalus mongholicus injection group were given by peritoneal from fourteen weeks old. Weight were measured fortnightly, and fasting plasma-glucose at 16,20 and 24 weeks. They were sacrificed at 24weeks of age respectively. The changes of the pathomorphology was observed.2. Tubular epithelial cells (HK-2) cell were divided into three groups:control,model and Puerarin injection combine Astragalus mongholicus injection group. When the cell adherenced, the model group's culture media addin 8ng/ml TGF-β1, the treat group's culture media addin 8ng/ml TGF-β1 and Puerarin inject ion combines Astragalus mongholicus inject ion. After 48h, BMP-7, BMP-7R and Smad 5 mRNA were detected in cell by RT-PCR, FN protein were detected in cell by immunohisto-chemical method.3. HK-2 cell were divided into three groups:control, model and Puerarin injection combine Astragalus mongholicus injection group. When the cell adherenced, the model group's culture media addin 8ng/ml TGF-β1, the treat group' s culture media addin 8ng/ml TGF-β1 and Puerarin injection combines Astragalus mongholicus injection.After 48h, BMP-7, BMP-7R and Smad 5 mRNA were detected in cell by RT-PCR, FN protein were detected by immunohisto-chemical method. Lip2000 group to deal with the same as SiRNA group without SiRNA. After 48h, BMP-7, BMP-7R and FN mRNA were detected by real-time PCR. Result1. KKAy mice developed obesity, hyperglycemia, after 14 weeks renal morphologic changes were discovered at 24 weeks of age, the model group demonstrated different degree of GBM thicking, the mesangial cell increased and the expansion of mesangial matrix was significant, and obvious vacuole in the renal tubular epithelial cells and hyal ine casts deposi t ion in tubular epithel ial cells, renal interstitial matrix accumulation.2. After the activation of TGF-(31, the appearance of HK-2 cell became DN change which the character of the cell became long and thin and there are abundant grains in the cytoplasm, we can also found huge confluent cell.3. Puerarin injection combines Astragalus mongholicus injection can improved the character of HK-2 and inhibited the expression of FN protein of HK-2 cell.4. Puerarin injection combines Astragalus mongholicus injection remarkably upregulated the expression of BMP-7, BMP-7R and Smad 5 mRNA.5. After Smad5 SiRNA interuption, Puerarin injection combines Astragalus monghol icus inject ion increased BMP-7R mRNA and do nothing wi th BMP-7 and FN mRNA.Conclusion1. Puerarin injection combines Astragalus mongholicus injection can ameliorate hyperglycemia of KKAy mice and ameliorate the pathology damage of kidney.2. Puerarin injection combines Astragalus mongholicus injection improved fibrosis of kidney.3. The possible mechanism of treating DN by Puerarin injection combines Astragalus mongholicus injection:up-regulating the expression of BMP-7 and BMP-7R mRMA to prevent its effect in induced and delaving DN4.The Tunction of BMP-7 which lessen kidney fibrosis maybe by BMP-7/Smad 5. passway.