The Effect Of Glomerular Mesangial Cells Proliferation And Cysteine Aspartic Acid Protease-3(Caspase-3) Protein Expression In IgA Nephropathy Model Rat Induced By Centella Asiatica Compound Mixture

Objective:Through animal experiments in rats,to observe the effect of glomerular mesangial cells proliferation and Cysteine aspartic acid protease-3(Caspase-3)protein expression in IgA nephropathy(IgA nephropathy,IgAN)model rat induced by Centella asiatica Compound mixture.to prove Centella asiatica Compound mixture can prevent and treat IgA nephropathy by inhibiting mesangial cell proliferation.to study its possible mechanism of Centella asiatica Compound mixture inhibiting the rat mesangial cells proliferation and provide a new direction for mechanism of mesangial cells proliferation and intervention research in IgAN.Methods:Fifty normal male SD rats,weight between 200±20g grams,were bred for one week firstly,and then randomly divided into five groups-normal control group(A)and IgAN rat model group(model group(B),treated group with Valsartan(C),,treated group with low dose of Centella asiatica Compound mixture(D),treated group with high dose of Centella asiatica Compound mixture(E)).Rats in IgAN rat model group using improved immune complex method by oral bovine serum albumin(BSA)+injected lipopolysaccharide(LPS)via caudal vein+subcutaneous injection of carbon tetrachloride(CC14+castor oil).Normal group were oral distilled water,injected equal volume of NS via caudal vein.From four weeks after model iducing,rats in group D,E and C were filled the stomach with Centella asiatica Compound mixture Chinese medicine decoction and valsartan suspension.while rats in group A and B 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 16th week.The 24h urinary protein excretion,Cr andBUN in blood were measured.Then renal morphology was observed by immunofluorescence,light microscopy(LM)and the expression level of Caspase-3 was examined by Western-blots.Results:1 24h urinary protein excretion:In the 10th and 16th week,group B was significantly higher than normal control group(P<0.01).In the 10th week,It was significantly lower in each treatment groups than group B(P<0.05),there was no statistical difference between experimental groups(P>0.05);In the 16th week,there was no statistical difference between group E and group A(P>0.05),group E was significantly lower than group D(P<0.01),there was no statistical difference between group C and group D(P>0.05),was significantly lower than group B(P<0.01),there was no statistical difference between 16th week and 10th week in group A(P>0.05).;It was significantly lower in 16th week than 10th week in each treatment groups and group B(P<0.05);2.Sreum Cr、BUN:there was no statistical difference between all groups(P>0.05).3.renal tissue immunofluorescence:we can see normal rat mesangial area without IgA immune complex deposition and IgAN model group glomerular mesangial with IgA immune complex deposition by immunofluorescence microscopy.IgAN rat model was successfully established.IgA fluorescence intensity of each treatment group decreased to varying degrees compared with the model group in the 10th and 16th weeks.4.Changes of rats’ renal pathology:Kidney tissue of rats after HE staining in LM:Glomerular structural integrity,good capillary loop open and mesangial matrix no obvious lesions in group A;Glomerular mesangial cell proliferation and mesangial matrix accumulation in group B;Each treatment group showed mild mesangial cell proliferation,mesangial matrix widening,mild disease compared with the model group.5.Effect of Caspase-3 expression in each group:Caspase-3 visible expression in normal renal tissue,and compared with normal group,the expression of the model group was significantly lower(P<0.01).Caspase-3 protein expression have varying degrees of increase after treatment with Centella asiatica Compound mixture and valsartan(compared with model group).Centella asiatica Compound on low and high dose group could significantly increase the expression of Caspase-3 protein(P<0.01 or P<0.05).Expression of Caspase-3 protein in a dose-dependent manner in a certain of Centella asiatica Compound mixture.Caspase-3 protein expression increased slightly in Valsartan group compared with the model group,but not significantly different(P>0.05).Conclusion:Using improved immune complex method by oral bovine serum albumin(BSA)+injected lipopolysaccharide(LPS)via caudal vein+subcutaneous injection of carbon tetrachloride(CC14+castor oil)can successfully established rat IgAN model.Each treatment group were having a role in reducing urinary protein.And to some extent inhibited IgAN rat mesangial cell proliferation,upregulate the expression of Caspase-3.