| Objective: In this topic research,in order to clarify the mechanisms of Centella asiatica in easing renal tubular interstitial fibrosis and provide theoretical basis for the clinical application of centella asiatica in preventing TIF,we observed the renal function and the expression of smad3 ? smad7 in the rats with Unilateral Ureteral Obstruction(UUO).Methods: Prepare the renal tubule interstitial fibrosis models by ligating the left ureter of 50 SD male rats,which are randomly divide into 5 groups-blank group,model group,Fosinopril group,high-dose Centella asiatica(HC)group and low-dose Centella asiatica(LC)group-10 rats were in each group.UUO rat model were established successfully in all rats except for the blank group.Treament groups were treated by intragastric administration at the same time every day: the dosage of centella asiatica were 5.6 mg/100g/day and 16.8 mg/100 g/day;the dosage of fosinopril sodium tablets was 1.67 mg/100g/day;Blank group and model group were given the same volume of distilled water.After 14 days,removed their renals to identify the impact of centella asiatica and fosinopril sodium tablets on the renal function and the expression of smad3 ? smad7.Detective method specifically as follows:(1)detect renal function by drawing blood from abdominal aorta;(2)HE staining was performed to observe the pathological changes of the obstructed kidney;(3)Masson staining was used to determine the degree of renal fibrosis;(4)Qualitative and quantitative analysis of the expression of Smad3 and Smad7 in renal tissue by immunohistochemistry;(5)PCR was used to determine expression of smad3?smad7 protein in the kidney tissues;(6)western blot is used to determine expression of smad3?smad7 protein in the kidney tissuesResults:(1)Rats' postoperative conditions: one model group rat died 1 days after the operation,one HC group rat and model group were died 4 days after the operation.(2)Compared with the blank group,SCR and BUN were higher in the UUO model,but there was no significant difference among the four groups(P >0.05);(3)Conventional HE and Masson staining: there were no significant pathological changes in blank group;UUO groups showed expansion of tubule cavity,deformation of tube wall,stiffness tubule epithelial cell falls off,atrophy and vacuoles degeneration is visible at the same time,cell proliferation and apoptosis,and atrophy of renal tubular basement membrane,deformation,lack of the part,renal interstitial infiltration of inflammatory cells,interstitial thickening of width,and extracellular matrix accumulation,collagen fibers increased(P <0.05);Compared with the model group,The treatment group was better than the model group but higher than the blank group(P<0.05);(4)Immunohistochemical results:(1)immunohistochemical staining results about smad3 in the kidney tissue: blank group showed a bit tan dyeing,which were weakly positive expression of the protein and confined to the renal tubules,occasionally seen in glomeruli.Compared with model group,Fosinopril group,LC group and HC group could significantly inhibit the generation of smad3(P<0.05),and the effect depended on the dose.(2)immunohistochemical staining results about smad7 in the kidney tissue : blank group showed a bit of tan dyeing which was mainly expressed in glomerular and tubular epithelial cells.Compared with model group,Fosinopril group,LC group and HC group could significantly increase the generation of Snail1(P < 0.05),and the effect depended on the dose.(5)PCR detection results:(1)After 14 days in UUO operation,compared with model group,Fosinopril group,LC group and HC group could could significantly inhibit the generation of smad3(P < 0.05),and the effecte dpended on the dose;the effect of the HC group was similar as the fosinopril group.(2)After14 days in UUO operation,compared with model group,Fosinopril group,LC group and HC group could significantly increase the generation of smad7(P < 0.05),and the effect of the high dosage was similar as the fosinopril group.(6)western blots detection results:(1)compared wirh the blank group,the expression of Smad3 protein was significantly increased in other groups;compared with model group,Fosinopril group,LC group and HC group could could significantly inhibit the generation of smad3(P < 0.05).(2)compared wirh the blank group,the expression of Smad7 protein was significantly inhibit in other groups;compared with model group,Fosinopril group,LC group and HC group could significantly increase the generation of smad7(P < 0.05).Conclusion:(1)Centella asiatica particles can inhibit expression of smad3 protein capacity;promote the expression of smad7 protein capacity,which suggested that the expression of Smad3 and Smad7 could delay the progression of the renal tubulointerstitial fibrosis after unilateral ureteral obstruction;(2)Centella asiatica particles affect smad3? smad7 are positively related with dosage,and the role of centella asiatica high-dose group is similar as the fosinopril group. |
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