Effect Of Brucine On Expression Of Podocyte Molecule Nephrin、Podocin With Heymann Nephritis

Objective To explore the therapeutic effect and mechanism of brucine on membranous nephropathy using passive Heymann nephritis(PHN)in rats,through the study of brucine on PHN rat podocyte molecule nephrin、podocin expression.providing experimental basis for clinical application.Methods The Methods Ninety-eight normal male SD rats,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 Fxl a 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 six groups-treated group with brucine in group A、brucine in group B、brucine in groupC、brucine in groupD,treated group with ramipril(E)and model group(F).From nine days after model iducing,rats in group A,B,C and D were administered intraperitoneally with brucine by 5mg/(kg.d),10mg/(kg.d),15mg/(kg.d)and 30mg/(kg.d),rats in group E were administered intragastrically with ramipril suspension by 5mg/(kg.d),while rats in group F 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 7th and 14th day,the remaining rats were sacrificed in the 21 th day.The 24h urinary protein excretion,Cr,ALB,WBC,ALT and AST in blood were measured.Then renal morphology was observed the expression level of IgG was examined by immunofluorescence and the expression level of nephrin、podocin was examined by immunohistochemistry.Results 1.Changes of rats’ blood and urine indices:(1)24h urinary protein excretion:In the 7th,14th,21th day,all experimental groups was significantly higher than normal control group(P<0.05).In the 7th day,there was no statistical difference between all experimental groups and group F(P>0.05).In the 14th day,group C、D were significantly lower than group F(P<0.05),there was no statistical difference between the otherl experimental groups(P>0.05);In the 21th day,there was statistical difference between all experimental groups and group F(P<0.05);there was statistical difference between all brucine groups and group E(P<0.05),there was no statistical differenc in brucine groups(P>0.05),With the increase of concentration increased,group D decreased most significantly.(2)Serum ALB level:In the 7th,14th,21th day,all experimental groups was significantly lower than normal control group(P<0.05).In the 7th day,here was no statistical difference between all experimental groups(P>0.05).In the 14th day,groupC、D was significantly higher than group F(P<0.05).there was no statistical difference between the otherl experimental groups(P>0.05);In the21th day,brucine groups were significantly higher than group F(P<0.05).there was no statistical difference between group E and group F(P>0.05).With the increase of concentration increased,group D increased most significantly.(3)Sreum Cr,ALT,AST level and blood WBC level:In the 7th,14th,21th day,there was no statistical difference between all groups(P>0.05).2.Changes of rats’ renal pathology:(1)Immunofluorescence IgG expression level:In the 7th day,there was no statistical difference in all experimental groups(P>0.05);In the 14th day,groupC、D was significantly lower than group F(P<0.05),there was no statistical difference between the otherl experimental groups(P>0.05);there was statistical difference between groups E and groupC、D(P<0.05);In the21th day,brucine groups were significantly lower than group F(P<0.05).there was no statistical difference between group E and brucine groups(P>0.05).(2)Immunohistochemistry nephrin、podocin expression level:In the 7th,14th,21 th day,all experimental groups was significantly lower than normal control group(P<0.05).In the 7th day,there was no statistical difference in all experimental groups(P>0.05);In the 14th day,the nephrin expression of groupC was significantly higher than group F,the podocin expression of groupC、D was significantly higher than group F(P<0.05),there was no statistical difference between the otherl experimental groups(P>0.05);the21tth day,brucine groups were significantly higher than group F(P<0.05),there was no statistical difference between group E and brucine groups(P>0.05).With the increase of concentration increased,group D increased most significantly.Conclusion(1)Brucine could reduce PHN rat’s proteinuria,may improve the level of ALB in PHN rats.(2)Brucine had effect on PHN rat kidney IgG and nephrin、podocin deposition,its can reduce the deposition of IgG;(3)upregulation of immunohistochemistry of nephrin、podocin protein expression,protect podocytes.(4)Therapeutic dose of brucine for PHN rat was safe.