| Objective:By building rat model with chronic renal failure(CRF)to study derivative of prostacyclin(PGI2) possible effect on expression of partial kidney's renin-angiotensin system(RAS) key factor:angiotensin ?(Ang ?), angiotensin(1-7) [Ang-(1-7)],angiotensin-converting enzyme 2(ACE2), angiotensin ? receptor type 1(AT1) in CRF rat model.to explore PGI2 possible protection mechanism in the pathogenic process of CRF.Method:1.Building 5/6 nephrectomy rat model of chronic renal failure.Normal rats were randomly divided into three groups:control group(aceept sham operation),CRF model group,CRF model with the derivative of PGI2 treatment grounp.10 rats in each group.dectected their Scr,BUN 5 weeks later,observed pathologic change from random one in each group in renal tissue pathological section staining to prove whether animal model was established successfully.treatment group receive beraprost sodium(BPS)treatment by gastric infusion at twice for 4weeks 0.6mg/kg/d, control group,model group receive equal distilled water as substitution.the rats of the three groups were killed 9 weeks later to observe the following indicators:1. 24 h total urinary protein.serum creatinine,usea nitrogen before and after intervention.2.Three groups of rat kidney tissue by hematoxylin and eosin(HE) staining and Masson trichrome staining(Masson) pathology3. q-RT PCR,Western blot,Immunofluorescence were used to detect ACE2,AT1 gene expression level and protein concentration in renal tissue4.Enzyme linked immunosorbent assay adsorption method(ELISA) were used to detect the concentration of Ang ? and Ang-(1-7) in renal tissue,the concentration of ACE2, Ang ?, ang(1-7), AT1 in serum.Results:1.Establishment and verification of an animal model of CRF:renal function:inCRF model group,Scr,BUN,24 h total urinary protein increased signifiantly,Renal pathological tissue changes: glomerular mesangial cell hypertrophy, tubule interstitial fibrosis with inflammatory cell infiltration,protein cast in renal tubles could be seen.After BPS treatment for 4 weeks, the glomerular mesangial cell proliferation,renal tubular interstitial fibrosis and inflammatory cell infiltration in the treatment group were significantly reduced compared with the model group.2.During the entire experiment, 24 h urine protein,Scr,BUN did not change significantly in the control group; compared with the control group, 24 h urine protein,Scr, BUN were increased(P <0.05) in Model group(5weeks)and treatment group( 5weeks); compared with the model group(5 weeks), at 9 weeks Scr, BUN decreased(P <0.05), 24 h total urinary protein was increased(P <0.05); after BPS gavage for 4 weeks( 9 weeks), compared with treatment group(5 weeks), 24 h urinary protein was decreased(P <0.05), and Scr, BUN levels did not change significantly.3.At 9 weeks, Western blot, q-RT PCR and immunofluorescence results showed: compared with the control group, ACE2 in kidney tissue of rats in the Model group, the expression of lowered(P < 0.05), the AT1 expression increased(P < 0.05).Compared with the Model group, the expression of ACE2 was up regulated(P<0.05),and the expression of AT1 was down regulated(P<0.05) in the treatment group.4.4.ELISA results:Renal tissue :Compared with the control group, the Concentration of Ang ? of the rats in 9 weeks Model group increased significantly(P < 0.01),the concentration of Ang(1-7) was decreased signficantly(P < 0.01); Compared with the 9 weeks Model group,the concentration of Ang ? in treatment group in 9 weeks was lower(P<0.05),the concentration of Ang(1-7) in treatment group was significantly higher(P<0.01).Serum: control group, Model group, at 5 weeks, 9 weeks respectively,the concentrations of Ang ?,Ang(1-7),ACE2 and AT1 did not change significantly(P>0.05). compared with the blank group, in Model group(5weeks), treatment group(5weeks),the concentrations of Ang ?, AT1 were increased(P <0.05), the concentrations of Ang(1-7),ACE2 were decreased(P <0.05), in which the Modelgroup, the treatment group phase had no significant difference(P> 0.05); compared with the treatment group(5weeks), after the BPS intervention for 4 weeks(9 weeks)in treatment group,the concentration of Ang ? was decreased(P <0.05), the concentration of Ang(1-7) was increased in( P <0.05), while there is no significant change in the concentrations of AT1, ACE2(P> 0.05);Conclusion:Prostacyclin derivatives can delay the progression of CRF rat model,which may play a role in regulating renal renin-angiotensin system key factor of Ang ?, ACE2 and angiotensin(1-7), AT1,also may reduce urinary protein to achieve it. |
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