| Objective: To explore the effects of yiqiyangyinhuoxue prescription on transientreceptor potential canonical6(TRPC6) and RhoA in kidney tissue of DN,and therelevance of reducing the urine protein,delaying the progress of renalpathology.Expound the effect of yiqiyangyinhuoxue prescriptions on DN and itseffective mechanism,thus providing experimental evidence for widespread clinicalapplication of the prescription.Method: Prepare diabetes model with intraperitoneal injection ofstreptozotocin,choosing clean level60male SD rats, adaptive feeding for aweek,fasted for12h,were randomly divided into:Model group,Blank group,Diovangroup, High dose group and Low dose group,each group has12rats.Modeling groupswere given45mg/kg disposable intraperitoneal injection of streptozotocin,blank groupwas given the same dose of sodium citrate buffer.All rats were housed separately inthe same animal room,a standard diet.After feeding72hours,continuous monitoring ofblood glucose,urine glucose,urine protein,urine,blood glucose <16.7/mmoL, urineglucose in++(inclusive++), negative urine protein, urine volume was less thannormal group150%for modeling failure.After modeling success,model group andblank group were fed with0.9%saline solution (1ml/100g d),diovan group weregiven medicine according to1.67mg/200g d,high dose group, low dose group weregiven medicine according to the adult dose of12.5times,6.25times.Continuousgiving medicine for6weeks.One day before the end of the experiment,usingmetabolic cages to collect24h urine.When the rats were sacrificed, collected bloodfrom abdominal aortic,then take kidney tissue for pathological examination.Observethe general condition of the rats.Contrast changes in hypertrophy index,24h urinaryprotein quantitative,biochemical markers(inclusive Scr, BUN, ALB, ALT, TC, TG,FBG).Treat kidney tissue with HE staining,PAS staining,observe changes of renalpathological by light microscopy,and idetect expression of TRPC6and RhoA in the kidney tissues by mmunohistochemical staining.Result:1. Improve the general condition of ratsThe general condition of the Blank group of rats to maintain good,while modulegroups of rats is rough, without burnish, depression, irritability, independent activitydecreased significantly, loose stoo,not forming, diet and urine volume increasedobviously, significantly reduce weight.After drug intervention, high dose group, lowdose group and Diovan group compared with model group rats fur burnish, less waterquantity, other conditions have improved.2. The effects on experimental rat kidney pathological histologyThe naked eye:Blank group rat kidney form, such as the size of a broad bean, darkyellow, the surface is smooth, coated without adhesion, easy tear;Model group ratskidney volume obviously increased, surface and slightly are pale, cortex medullacolor deepened, with a clear demarcation between the cortex and coated withadhesion.HE staining and PAS staining:The structure of glomerular was integrated inBlank group rats, and there was no significant difference on mesangialmatrix,capillary endothelial cells and renal tubular epithelial cells.The renal tissue ofthe Model rats were glomerular hypertrophy, mesangial region widen, matrixincreased, diffuse thickening of capillary basement membrane,tubulointerstitialappear infiltration of inflammatory cells, with glycogen storage in glomerular matrixand under capillary endothelium.Chinese medicine high dose group, low dose groupand Diovan group rat kidney lesions were alleviated through drug intervention,.3.The compare of body weight, kidney weight, hypertrophy index (kidney weight/body weight) of rats in each groupsThe body weight of rats in the Blank group and Model group differences, but nosignificant differences in kidney weight. Comparison of hypertrophy index as follows:normal control group and Model group have significant difference (P<0.01); there isobvious difference between Model group and Diovan group (P<0.01), there are alsodifferences with High, Low dose group (P<0.05); there were statistically significantdifferences in Diovan group and high,Low dose group (P<0.01);or there aredifferences between the High, Low dose group (P<0.01).4.Effect on biochemical parameters in experimental ratsThere are no significant difference about BUN, Scr and ALT in each group rats,(P>0.05). A comparison between ALB in each group: the difference between the Blank group and Model group is statistically significant(P<0.01); Diovan group (18.28±0.90) and Model group (16.99±0.69) differences (P<0.05), and High dose group(21.80±1.16), Low dose group (20.81±0.99) difference more statistically significant(P<0.01); there was no significant difference in High, Low dose group (P>0.05). TC:the Blank group (1.39±0.26) compared with Diovan group, Model group, Low dosegroup, there was significant difference (P<0.01), and High dose group (1.62±0.09)was not significantly different (P<0.05), Diovan group (2.08±0.20) and Model group(2.09±0.22) no difference statistical significance (P>0.05). TG: there was significantdifference compared with Model group and Blank group (1.12±0.38)(P<0.01orP<0.05), Diovan group (3.32±0.37) and Model group (3.60±0.68) had no significantdifference (P>0.05).FBG:The difference between the Blank group and Model groupwas statistically significant(P<0.01);Model group and treatment groups haddifference(P<0.01or(P<0.05);the difference between Diovan group and High, Lowdose group was statistically significant(P<0.01);there was no significant difference inHigh, Low dose group (P>0.05).5.Comparison of rat24h urinary protein quantitative24h urinary protein quantitative of Blank group rats (0.0371±0.0166) and High dosegroup (0.0553±0.0075) there was no significant difference (P>0.05), and the othergroup (Diovan group, High dose group, Low dose group) had a statistically significantdifference (P<0.01); Diovan group (0.0834±0.0167) and Low dose group (0.0802±0.0121) had no significant difference (P>0.05), and there are differences and Highdose group (P<0.01).6.Expression of TRPC6and RhoA in renal tissues of rats in each groupExpression of TRPC6in podocyte cell membrane, expression of RhoA was mainlydistributed in the cell membrane,also cytoplasm. Compared with the Blank group,Model group the expression of TRPC6and RhoA increased significantly, thedifference was statistically significant (P<0.01); the expression quantity of TRPC6and RhoA in Model group is most, and other modeling group (Diovan group, highdose group, low dose group) had a statistically significant difference (P<0.01); theexpression of TRPC6and RhoA had no difference in the High, Low dose group(P>0.05);Conclusion:1. Yiqiyangyinhuoxu prescriptions can decrease the urine protein,correcthypoproteinemia,glucose metabolism and improve lipid metabolism; 2. Yiqiyangyinhuoxu prescriptions can inhibit the DN rat glomerular capillarybasement membrane thickening, mesangial matrix proliferation and cells of thepathological changes, delay the progress of renal pathology.3. Yiqiyangyinhuoxue prescriptions may influence the expression of TRPC6andRhoA in renal tissue,which play a role above.. |
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