| Diabetic nephropathy is one of the most common microvascular complications of diabetes and is also the most dangerous factors leading to end-stage renaldisease(ESRD),so we need effective and innovative treatment to prevent the pathogenesis of diabetic nephropathy and chronic kidney disease.It has been confirmed that oxidative stress and inflammatory response processes are important factors leading to renal damage in diabetic nephropathy.Therefore,by inhibiting oxidative stress and inflammatory response can be used as a new target for treatment.Clinical studies have confirmed the effectiveness of single herbs in the treatment of diabetic nephropathy.In this study,we used the addition and subtraction of the treatment of diabetic nephropathy to conduct an in-depth molecular mechanism study of how did chinese medicine delay the pathogenesis of diabeticnephropathy.Part ?.Establishment of rat model of diabetic nephropathy and observation of histomorphologyObjective:all rats of diabetic nephropathy model were established by with unilateral nephrectomy and intraperitoneal injection of streptozotocin(STZ)induced production.By observing the general condition of the rats and pathological changes of renal tissue to verify that GLQM can treat diabetic nephropathy and can delay the further progress.Methods:100 SD rats were purchased and adapted for one week,except those in the normal group,all rats of diabetic nephropathy model were established bywith unilateral nephrectomy and intraperitoneal injection of streptozotocin(STZ)induced production,After 72 hours of STZ injection,the random blood glucose?16.7 mmol L-1 was determined that the model was succeed.Then rats were randomlydivided into five groups:the model group,and the four treated groups treated with low dose of GLQM-L,medium dose of GLQM-M,high dose of GLQM-H,positive medicine(valsartan),The treatments were given via gastrogavage everyday starting from the 4th week of modeling,groups treated with high,medium and low dose of GLQM were given via gastrogavage to 5.6g·kg-1,2.8g·kg-1,1.4g·kg-1,positive drugs(valsartan)group were given via gastrogavage to 4.8×10-3g·kg-1,the normal group and model group were given 2.8g·kg-1distilled water,once a day for 12 weeks.At the 4th and 12th week of treatment,the FBG values were measured.After 12 hours of the last administration,the rats were sacrificed by cervical dislocation.we observed the sign of rats and therat glomerular and the rats were sacrificed by routine paraffin sections and subjected to HE and PAS,the remaining kidney tissue was stored in liquid nitrogen for use.Results:The hair of the normal group was evenly ditributed,and the color wasnormal and the activity was normal.The hair of the model group were sparse,and the hair color was yellowish and they had the symptoms of drinking,eating,polyuria,weight loss and mental malaise.In the course of the experiment,the distribution of coat color and hair was improved compared with the modelgroup,and the body weight had a certain degree of increase compared with the model group.Compared with the DN model group,the pathological changes of the renal tissue were improved in different treatment groups.Conclusions:GLQM was effective in repair and regeneration of the damaged tissue.Part ?.Effect of GLQM on Inflammatory Factors in Rats with Diabetic NephropathyObjective:To explore intervention of GLQM on Inflammatory Factors in rats with diabetic nephropathyMethods:Take the first part of the stored kidney tissue in the liquid nitrogen,then we used the homogenate with tissue to determination the content the basic fibroblast growth factor(bFGF),Insulin-like Growth Factor(IGF),Monocyte ChemotacticProtein-1(MCP-1):in renal issues of rats by ELISA.Results:The bFGF,IGF,MCP-1 in renal tissues of rats with model were higher than the nomal group,there were significant difference among the model group and the norml group(P<0.01).The bFGF in renal tissues of rats with GLQM-H and GLQM-M,and MCP-lin renal tissues of rats with GLQM-L and bFGF IGFand MCP-1 in renal tissues of rats withvalsartan were lower than the model group,there were significant difference among them;the bFGF in renal tissues of rats with GLQM-L,and IGF in renal tissues of rats with GLQM-M,and MCP-1 in renal tissues of rats with GLQM-H were lower than the model group,there wassignificant difference between two(P<0.05).Conclusions:GLQM can decrease the expreion of bFGF,IGF,MCP-1 in renal tissues of rats,improve the renal microcirculation,be effective in repair and regeneration of the damaged tissue.Part ?.Mechanisms of Gualou Qumai Tang(GLQM)on p38 MAPK signaling pathway in rat diabetic nephropathyObjective:To explore intervention of GLQM in preventing the development of diabetic nephropathy(DN)through p38 MAPK signaling pathway methodsMethods:Take the first part of the stored kidney tissue in the liquid nitrogen,then we extracted the rat kidney protein and RNA.The protein expression of p-p38 MAPK?p-CREB?FN in renal tissue we also examined using western blot method.The mRNA expressions FN in renal tissue we examined using RT-PCR method.Results:The protein expression of p-p38 MAPK?p-CREB?FN in renal tissues with model were higher than the normal group(P<0.05 or P<0.01).The mRNA expression of FN in renal tissues of rats with GLQM were lower than the model group(P<0.05orP<0.01);the protein expression of p-p38 MAPK?p-CREB?FN in renal tissue of rats with GLQM were lower than the model group(P<0.05 or P<0.01).Conclusions:GLQM can reatrain the activation of p38 MAPK signaling pathway and be effective in repair and regeneration of the damaged tissue,it is also slow down the process of DN.Part IV.Mechanisms of Gualou Qumai Tang(GLQM)on TGF-?1/Smad signaling pathway in rat diabetic nephropathyObjective:aTo explore intervention of GLQM in preventing the development ofdiabetic nephropathy(DN)through TGF-pi/Smad signaling pathway methodsMethods:Take the first part of the stored kidney tissue in the liquid nitrogen and the spare slices,then we extracted the rat kidney protein and RNA,The protein expression of smad2 and smad3 were immunohistochemically investigated.The mRNA expressions TGF-?1?Smad2?Smad7 in renal tissue we examined using RT-PCR method.The protein expression of Smad2?Smad3?Smad7 in renal tissue we also examined using western blot method.Results:The mRNA expression of TGF-?1?Smad2 in renal tissues of rats with model were higher than the normal group,the mRNA expression of Smad7 in renal tissues of rats with model were lower than the normal(P<0.01).The protein expression of Smad2 and Smad3 in renal tissues with model were higher than the normal group,the protein expression of smad7 in renal tissue with model were lower than the normal group(P<0.01).The mRNA expression of TGF-?1?smad2 in renal tissues of rats with GLQM were lower than the model group,the mRNA expression of Smad7 in renal tissues of rats with GLQM were higher than the model group(P<0.01).The protein expression of Smad2 and Smad3 in renal tissues of rats with the model group,the protein expression of Smad7 in renal tissueof rats with GLQM were lower than the model group.Conclusions:GLQM can reatrain the activation of TGF-?1/smad signaling pathway and beeffective in repair and regeneration of the damaged tissue,it is also slow down the process of DN. |
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