Study Of Shenxiao Decoction On The Preventive Effect And The Mechanism Of Diabetic Nephropathy

Objective: Research the effect of Shenxiao decoction on the basicpathological changes of diabetic nephropathy rats’renal. Around the TGF-β1/Smad signal pathway, analyze Shenxiao decoction mechanism of action.Method: Healthy50male Sprague-Dawley (SD) rats weight200g±20gwere used in this study. After7days to adapt, randomly selected10as controlgroup (A group),40to create the diabetic model. Create the diabetic model byintraperitoneal injection of STZ, after72h, determination tail vein blood of themodel of glucose, higher than16.7mmol/L for successful diabetes model.Then randomly divided into three groups: model group (B group), irbesartangroup (C group), Shenxiao group (D group). Intragastric administration,Shenxiao group according to20g/kg·d, irbesartan group according to25mg/kg·d, model group and the control group were administrated withdistilled water,1times per day, lasted12weeks, free diet. In12weekendadmeasurement glucose; left24h urinary, determination of urinary albumin(mAlb) content; anesthesia animal by intraperitoneal injection of10%chloralhydrate. abdominal aortic blood, conservation the samples of serum todetection blood urea nitrogen (BUN), serum creatinine (Scr), cholesterol (TC),triglyceride (TG). The left renal tissue,4%paraformaldehyde solution, HE,PAS staining, to observe the change of renal pathology, transforming growthfactor-β1(TGF-β1), vascular cell adhesion molecular (VCAM) were detectedby immunohistochemistry. The right renal tissue; the cortex cut1mm3organization was fixed on the electron fluid, electron microscope observationof the ultrastructural changes of kidney; the rest stored in liquid nitrogen. ThemRNA of TGF-β1was detected by reverse transcriptase polymerase chainreaction(RT-PCR). Smad2/3, Smad7, connective tissue growth factor (CTGF),fibronectin (FN) were detected by Western blot.Result: 1The effect of Shenxiao decoction on the GluModel group, irbesartan group, shenxiao group were significantly higherthan the control group (P<0.05), between model group, irbesartan group,shenxiao group had no statistical significance (P>0.05), shenxiao group haddowntrend.2The effect of Shenxiao decoction on the24h mAlbModel group, irbesartan group, shenxiao group were significantly higherthan the control group (P<0.05), model group was significantly higher thanirbesartan group and shenxiao group (P<0.05), between irbesartan group andshenxiao group had no statistical significance (P>0.05).3The effect of Shenxiao decoction on the renal functionCompare the blood urea nitrogen (BUN) and serum creatinine (Scr),model group, irbesartan group, shenxiao group were significantly higher thanthe control group (P<0.05), model group was significantly higher thanirbesartan group and shenxiao group (P<0.05), between irbesartan group andshenxiao group had no statistical significance (P>0.05).4The effect of Shenxiao decoction on the blood lipidCompare the cholesterol (TC), model group, irbesartan group, shenxiaogroup were significantly higher than the control group (P<0.05), model groupwas significantly higher than irbesartan group and shenxiao group (P<0.05),between irbesartan group and shenxiao group had no statistical significance(P>0.05). The triglyceride (TG) had no statistical significance (P>0.05),shenxiao group had downtrend.5The effect of Shenxiao decoction on the pathologic changesHE staining: There was no change in control group. In model group, therenal pathological changes obviously, glomerular basement membrane (GBM)thickening, extracellular matrix (ECM) accumulation, proliferation ofmesangial cells; Irbesartan group and shenxiao group had thin lesions lowerthan in the model group, the difference was not obvious between irbesartangroup and shenxiao group. PAS staining: Control group renal glomerularstructure basically normal, PAS positive substance without significantly increased. In model group, renal pathological changes obviously, mesangialPAS positive substances increased obviously, red dye, glomerular basementmembrane (GBM) thickening, extracellular matrix (ECM) accumulation,proliferation of mesangial cells. Irbesartan group and shenxiao group wereslighter than in the model group. Electron microscope observation: Controlgroup glomerular basement membrane structure basically normal, uniformthickness, podocyte structure was normal, podocyte apoptosis, withoutshedding. Model group glomerular basement membrane (GBM) thickeningwas wide, the segment, a fusion of foot processes, loss of podocytes.Compared with model group, Irbesartan group and shenxiao group wereslighter, glomerular basement membrane (GBM) thickening was not obvious,the fusion of foot process less.6The effect of Shenxiao decoction on the VCAM, TGF-β1VCAM, TGF-β1of control group in glomerul, tubules have a smallamount of expression; expression levels of model group, irbesartan group,shenxiao group VCAM, TGF-β1was significantly higher than the controlgroup, model group showed a large number of expression in glomeruli, inendothelial cells, mesangial cells show high expression in contrast, matrixextracellular have scattered positive expression; expression levels of irbesartangroup and shenxiao group of VCAM, TGF-β1was lower than the model group,whole down regulated, part of glomerular had a higher expression, thedifferences between the two groups was not obvious.7The effect of Shenxiao decoction on the TGF-β1mRNAModel group, irbesartan group, shenxiao group were significantly higherthan the control group (P<0.05), model group was significantly higher thanirbesartan group and shenxiao group (P<0.05), between irbesartan group andshenxiao group had no statistical significance (P>0.05).8The effect of Shenxiao decoction on the Smad2/3, Smad7, CTGF, FNSmad2/3: model group was significantly higher than the control group,irbesartan group, shenxiao group (P<0.05), between control group, irbesartangroup, shenxiao group had no statistical significance (P>0.05). Smad7: irbesartan group, shenxiao group were significantly higher than the controlgroup and model group (P<0.05), shenxiao group was significantly higherthan the irbesartan group (P<0.05), between model group and control group nostatistical significance (P>0.05). CTGF and FN: model group, irbesartangroup, shenxiao group were significantly higher than the control group(P<0.05), model group, shenxiao group were significantly higher than theirbesartan group (P<0.05), model group was significantly higher than theshenxiao group (P<0.05).Conclusion1Shenxiao decoction has the function of decreasing24h mAlb, Scr andBUN, to protect the renal function.2Shenxiao decoction has the function of decreasing TC, glucose and TGdowntrend.3Shenxiao decoction has the function of reduce the renal pathologicaldamage, protect the renal function.4Shenxiao decoction has the function of reduce TGF-β1mRNA andTGF-β1, Smad2/3, increase Smad7. TGF-β1/Smad signaling pathway may beone of the mechanisms of shenxiao decoction effect for renal protection.5Shenxiao decoction has the function of decreasing VCAM, CTGF, FN,reduction of extracellular matrix (ECM), reduce renal fibrosis, protect thekidney.6Shenxiao decoction has the similar effect of the renal protective withirbesartan, moreover also has certain regulating blood lipid, blood sugar, bloodenvironment can be improved.