| Objectives To explore the effect of Shenshao oral liquid(Shenshao oral liquid) on diabetic nephropathy(DN) in rats with inflammation and fibrosis and mechanism.Methods In this study, Streptozotocin(HFD) + high fat diet(diet, STZ) was used to replicate the diabetic nephropathy(DN) model. 50 SPF Wistar rats in the experimental animal room adaptive feeding for 7 days were randomly divided into five groups, the control group(Control group), model group(DN group) and low dose group(Low dose group), middle dose group(Medium dose group), high dose group(High dose group), 10 rats in each group, the control group(CG) fed with normal feed, in addition to the control group(CG) and were fed with high fat diet(HFD) feeding, one month later, in addition to the control group(CG) and STZ(25mg/kg) of each group were given intraperitoneal injection in the rat tail, continuous measurement of blood glucose 2 times each time,more than 16.7mom/l for the successful modeling, until after the success of the model,each group according to the original feed, the low dose group(LDG) with Shenshao oral liquid(SSOL) perfusion(125mg/k/d), middle dose group(MDG)with(250mg/kg/d), high dose group(HDG) orally every day(500mg/kg/d), 8 weekslater Modle success. At the end of the experiment, two days before collecting 24 h urine of rats in all groups were measured by urinary protein in 24 hours microalbuminuria(UMA), collected morning each fasting rats placed in the cleaning of the metabolism cage, the next day the urine to volume of miscellaneous quality, sampling and delivery inspection department. Before the animals were sacrificed from aortic blood determination of serum creatinine(SCR),blood urea nitrogen(BUN), renal tissue samples were collected. By HE staining method light microscope observed under kidney cells in pathological changes, with Masson staining were used to observe the pathological changes of renal fibrosis, immune group and immunoblot(Western blot) was detected in renal tissue of rats with inflammatory factor of TLR4 and NF-k B and TGF-?1 protein expression changes.Results 1 General situation: the drinking,feeding,looking of the rats in control group was no abnormal; compared with the control group, the model group rats has high blood glucose,drinking more,eating more, fur has no lightand reduce,andthere is mental differences, less activity, slow movement, less urine in the rats of model group. Compared with the model group(DN), the medicine group There was significantly improved. 2 Determination results of Scr, BUN and UMA: at the end of the 8th week, the detection values of 24 UMA,Scr and BUN in model group were significantly higher than that in control group, and the difference is statistically significant(P < 0.01). The detection values of 24 UMA Scr and BUN in each drug group were significantly lower than than in the model group, the difference has statistical significance(P < 0.01). 3 renal pathological change: 8 weeks after treatment, compared with the control group, model group rat glomerular sclerosis is bigger, The model group rats glomerular sclerosis,basement membrane thickening and mesangial matrix are part of renal cysts become narrow.Compared with the model group(TMG), the drug group(DN) has less pathological changes and less inflammatory cells. 4 Masson staining:Compared with the control group, the rats in model group were more serious in renal tissue fibrosis.Compared with the model group, the renal tissue fibrosis of the rats in each drug group was lighter than the model group. 5Immunohistochemical display: Compared with the control group, the expression of TLR4, NF-k B, TGF-?1 protein in kidney tissue of model group rats was raised, the difference has statistical significance(P < 0.01), Compared with the model group, the expression of TLR4, NF-k B, TGF-?1 protein in renal tissue of each drug group was down regulated, and the difference is statistically significant(P <0.01). 6 Western blot results showed that: Compared with the control group model group(DN) rats kidney tissue TLR4,NF-k B,TGF-?1expression up regulation, and the difference is statistically significant(P < 0.01), Compared with the model group, the drug group was significantly lower, and the difference is statistically significant(P < 0.01).Conclusions 1 Shenshao oral liquid(SSOL) has anti-inflammatory and anti fibrosis effect,and there was a dose dependent relationship in a certain range. 2 Shenshao oral liquid may have anti-inflammatory and anti fibrotic effects by reducing TLR4 expression of and NF-k B and TGF-?1 protein and so as to protect the kidney. |
PDF Full Text Request