| Objective To explore the therapeutic effect and mechanism of brucine on membranous nephropathy using passive Heymann nephritis in rats,providing experimental basis for clinical application.Methods Ninety-eight normal male SD rats,weight between 120-150 grams,were bred for one week firstly,and then randomly divided into two groups-normal control group and PHN rat model group.Rats in PHN rat model group were injected sheep anti-rat Fxla serum(0.4ml/100g) while rats in normal control group were injected equal volume of NS via caudal vein.Rats getting successful modelling were then randomly divided into five groups-treated group with low dose of brucine(A),treated group with middle dose of brucine(B),treated group with high dose of brucine(C),treated group with ramipril(D) and model group(E).From nine days after model iducing,rats in group A,B and C were administered intraperitoneally with brucine by 5mg/(kg.d),10mg/(kg.d) and 15mg/(kg.d),rats in group D were administered intragastrically with ramipril suspension by 5mg/(kg.d),while rats in group E were administered ingastrically with equal volume of NS everyday.During the period of the experiment,rats took food and water freely.Five rats were sacrificed in each group in the 10th and 20th day,the remaining rats were sacrificed in the 30th day.The 24h urinary protein excretion,Cr,ALB,WBC,ALT and AST in blood were measured.Then renal morphology was observed by light microscopy(LM) and transmission electron microscope(TEM),the expression level of IgG was examined by immunofluorescence and the expression level of membrane attack complex was examined by immunohistochemistry.Results 1.Changes of rats’ blood and urine indices:(1)24h urinary protein excretion:In the 10th,20th,30th day,group E was significantly higher than normal control group(P<0.05).In the 10th,30th day,there was no statistical difference between experimental groups and group E(P>0.05),there was no statistical difference between all experimental groups(P>0.05).In the 20th day,group C was significantly lower than group E(P<0.05),there was no statistical difference between experimental groups(except group C) and group E(P>0.05),there was no statistical difference between all experimental groups(P>0.05).(2)Serum ALB level:In the 10th day,group E was significantly lower than normal control group(P<0.05),there was no statistical difference between experimental groups and group E(P>0.05),there was no statistical difference between all experimental groups(P>0.05).In the 20th,30th day,there was no statistical difference between all groups(P>0.05).(3)Sreum Cr,ALT,AST level and blood WBC level:In the 10th,20th,30thday,there was no statistical difference between all groups(P>0.05).2.Changes of rats’renal pathology:(])The renal morphology of model group rats by LM and TEM in the 30th day:LM showed glomerular basement membrane thickening,spike formation and glomerular basement membrane vacuolation degeneration.TEM showed many electron dense with various sizes deposited in glomerular basement membrane and under podocytes,glomerular basement membrane uneven thickening,spike formation and diffuse effacement of podocyte foot process.(2)Immunofluorescence IgG expression level and immunohistochemistry membrane attack complex expression level:In the 10th,20th,30th day,there was no statistical difference between experimental groups and group E(P>0.05),there was no statistical difference between all experimental groups(P>0.05).Conclusion PHN rat model was a good human MN model with short modelling time and high success rate.Its clinical manifestations and clinical features were similar to human beings.Brucine could reduce PHN rat’s proteinuria,had no effect on PHN rat kidney membrane attack complex and IgG deposition,its mechanism of reducing PHN rats’ proteinuria needs further reseach.Therapeutic dose of brucine for PHN rat was safe. |
PDF Full Text Request