| Objective: To observe the Yiqi Huayu Qingre formula and its compositions' effect on urinary red blood cell count,24 hours urine protein,blood biochemical indexes,renal pathology changes of experimental rats model with IgA nephropathy,besides,we will also research nephrin's and podocin's quantity and distribution in glomerular epithelial cells to investigate the mechanism of Yiqi Huayu Qingre formula and its compositions' curative effect on IgA nephropathy.Methods: Wistar male rats of SPF grade were randomized allocation to 11 groups : blank group,model group,Yiqi group,Huayu group,Qingre group,herb compound group,Tripterygium wilfordii group,high dose of Yiqi group,high dose of Huayu group,high dose of Qingre group,high dose of herb compound group;Using bovine serum albumin(BSA)as antigen filling the stomach for every two days to stimulate the rats,injecting carbon tetrachloride(CCl4)to rats' neck skin weekly,injecting lipopolysaccharide as immune adjuvant at the 6th week through tail vein to make IgA nephropathy model.Count the urinary red blood cell and test the amount of 24 hours urine protein.At the same time,the expression of nephrin and podocin protein in glomerular endothelial cells were observed by immunohistochemical staining,and the staining intensity was analyzed.Results: 1 In the urine red blood cell count and 24-hours urine protein aspects1.1 From the ending of the 6th week,the rats except blank group began to appear hematuria,proteinuria,and showed continuous progression,but there was no gross hematuria.1.2 At the 10 th week,urinary red blood cell's amount and 24 hours urine protein quantitation of Tripterygium wilfordii group and high dose of compound group were decreased significantly as compared with model group,the difference was statistically significant(P < 0.01);the rest groups of rats' hematuria and proteinuria remained elevated,and had no significant difference compared with the model group(P>0.05).1.3 At the end of the 12 th week,the urinary red blood cell's amount and 24 hours urine protein quantitation of every treatment group were decreased significantly compared with the model group,the difference is statistically significant(P < 0.01).Both the high dose group and Tripterygium wilfordii group had no significant difference compared with the blank group,P > 0.05.There was no difference between the two groups,P > 0.05,which showed that the high dose compound group had the strongest effect in this experiment.About the two indexes,the compound group was lower than the Huayu group and the Qingre group,the Huayu group and the Qingre group were lower than the Yiqi group,high dose of the compound group was lower than the high dose of Huayu group and high dose of Qingre group,the high dose of Huayu group and high dose of Qingre group were lower than high dose of Yiqi group,high dose was lower than the nomal dose of each group.2 Blood biochemical resultsBlood biochemical indexes were detected in experimental rats in three different time points and there was no difference between every two groups of all groups in indexes such as ALB?TP?TCH?TG?Cr?BUN(all P>0.05);About ALT and AST in rats,The Tripterygium wilfordii group was significantly higher than other groups,the difference was statistically significant,while among other groups,there's no statistical difference.All these indicated that different intervening factors had no influence on ALB?TP?TCH?TG?Cr and BUN in blood,but Tripterygium wilfordii had certain effect on experimental rats' liver function.3 Renal pathology The glomerular volume of IgAN model rats increased slightly,mesangial cells were mild to moderate hyperplasia,accompanied with mild to moderate enhancement of mesangial matrix.The pathologic findings were mainly distributed from grade II to grade III.We could see that the glomerular mesangial area and the capillary wall had strong granular or agglomerate IgA compound deposited,the fluorescence intensity was mainly degree(+ + +).In the treatment groups,the renal pathology performance of Yiqi group was the same as that of the model group,the pathologic findings were mainly distributed from grade II to grade III,the fluorescence intensity was mainly degree(+ + +).Except Yiqi group,the other groups' pathological changes were relieved compared with the model group,Presented mesangial cells' mild hyperplasia,companied with mesangial matrix's slight widen.The degree of pathological changes fluctuated between grade I and grade II.Under immuno fluorescence microscope,the expression intensity of IgA in the glomerular mesangial area was significantly lower than that in the model group,show a weak positive,Mainly in the segmental IgA deposition,the fluorescence intensity was in the scope of degree(+)to(+ + +).4 Expression of nephrin and podocin protein The expression of nephrin?podocin protein of the high dose of compound group was higher than the high dose of Qingre group and the high dose of Huayu group,the differences were statistically significant P<0.01,indicating that the compound group's effect was higher than the high dose of Qingre group and the high dose of Huayu group.The expression of podocin protein of the Qingre group and the Huayu group was higher than the high dose of Yiqi group and the Yiqi group,the difference was statistically significant P<0.01,but there was no difference between the two groups.The expression of podocin protein of the high dose of Yiqi group and the Yiqi group is weaker,but better than the model group,and there was statistical difference,P<0.01,but there was no difference between the two groups.Which indicating that the effect of the high dose of Yiqi group and the Yiqi group were the least,weaker than the Qingre group and the Huayu group.Conclusion:(1)The Yiqi Huayu Qingre formula can significantly improve the IgA nephropathy rat hematuria,proteinuria,the Yiqi group's effect was the least,weaker than the compound group?the Huayu group and the Qingre group,the high dose of compound group's effect was the best.(2)The improvement of hematuria and proteinuria of the Yiqi Huayu Qingre formula in IgA nephropathy rats may be realized by regulating the expression of nephrin and podocin protein.(3)The effect of the complex high group appeared early,with good effect and little side effect,which is worth of clinical application. |
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