| Objective: To investigate the protective effects of Yiqi-Yangyin-Qingre-Lishi Recipe on Ig A Nephropathy and its possible mechanism, in order to provide experiment foundation for clinical treatment of Ig A nephropathy, by observing the effects of the Chinese medicine on the 24-hour proteinuria, serum creatinine, blood urea nitrogen, albumin and the expression of matrix metalloproteinases-9(MMP-9) and matrix metalloproteinases inhibitor-1(TIMP-1).Methods:40 healthy male Sprague-Dauley rats were randomly divided into control group(n=10), model group(n=10), benazepril group(n=10)and Chinese traditional medicine group(n=10). The Ig AN model was established by administration of bovine serum albumin(BSA), lipopolysaccharide(LPS), and carbontetrachloride(CCl4) after adaptive fed a week except control group rats. Immunogen BSA was intragastrically administered at 400 mg/Kg once every two days for 6 consecutive weeks and 0.1 m L of CCl4 in 0.5 m L of castor oil was given once weekly for 9 consecutive weeks. At the 6th and 8th week, 0.05 mg of LPS was injected through the tail vein. For the control group, 4 m L/Kg distilled water was used to replace the BSA, 0.6m L of castor oil was used to replace castor oil and CCl4, and saline was used to replace LPS at the 6th and 8th week. From 10 th week, benazepril group and Chinese traditional medicine group were treated with benazepril and Chinese traditional medicine; control group and model group were treated with distilled wate for 8 weeks. Twenty-four-hour urine of all rats with metabolism cages was collected before model establishment and at the end of 7th week, 9th week, 13 th week and 17 th week to determine 24-hour proteinuria. All rats were killed to obtain the serum and the renal tissues. Scr, BUN and ALB were assayed with the automaticbiochemistry analyzer. Sections of kidneys were stained by hematoxylin and eosin(HE) and periodic acid-silver metheramine(PASM), dentified by immunofluorescence of Ig A. The expression of MMP-9 and TIMP-1 in renal tissue was measured by Immunohistochemistry, RT- PCR.Results:1 24-hour proteinuria of four groups at the same timeThere was no significant difference in four groups before model establishment(P>0.05).At the end of the 7th week, compared with control group, 24-hour proteinuria of rats in model group, benazepril group and Chinese traditional medicine group was significantly higher(P<0.05); there was no significant difference among the model group, benazepril group and Chinese traditional medicine group(P>0.05).At the end of the 9th week, compared with control group, 24-hour proteinuria of rats in model group, benazepril group and Chinese traditional medicine group was higher significantly(P<0.05); there was no significant difference among the model group, benazepril group and Chinese traditional medicine group(P>0.05).At the end of the 13 th week, compared with control group, 24-hour proteinuria of rats in model group, benazepril group and Chinese traditional medicine group was higher significantly(P<0.05); compared with model group, 24-hour proteinuria of rats in benazepril group and Chinese traditional medicine group was significantly lower(P<0.05); there was no significant difference between benazepril group and Chinese traditional medicine group(P>0.05).At the end of the 17 th week, compared with control group, 24-hour proteinuria of rats in model group, benazepril group and Chinese traditional medicine group was significantly higher(P<0.05). Compared with model group, 24-hour proteinuria of rats in benazepril group and Chinese traditional medicine group was significantly lower(P<0.05); there was no significant difference between benazepril group and Chinese traditional medicine group(P>0.05).2 Scr, BUN and ALB of four groupsThere was no significant difference among four groups(P>0.05).3 The pathomorphology changes of kidney3.1 Light microscope observationControl group rats had normal kidney tissue structure. Model group rats were characterized by increased number of cells in the glomerulisevere, hyperplasia of mesangial matrix and cells. Benazepril group and Chinese traditional medicine group were characterized by mild hyperplasia of mesangial matrix and cells.3.2 Immunofluorescence of Ig AThere was no immune complex deposition in glomerular mesangial area in control group rats. The green immune complex deposited granularly along the glomerular mesenterium in model group rats. The degree of immune complex deposition in benazepril group and Chinese traditional medicine group was lighter than that in the model group.4 Immunohistochemical result of the expression of MMP-9 and TIMP-1 in four groupsMMP-9 was highly expressed in renal cortex of rats in control group.Compared with control group, the expression of MMP-9 in model group, benazepril group and Chinese traditional medicine group was remarkably decreased(P<0.05); compared with model group, MMP-9 in benazepril group and Chinese traditional medicine group had higher expression(P < 0.05). There was no statistical difference between two treatment groups(P>0.05).TIMP-1 in renal cortex of rats in control group had low expression. Compared with control group, the expression of TIMP-1 in model group,benazepril group and Chinese traditional medicine group were remarkably increased(P<0.05); compared with model group, TIMP-1 in benazepril group and Chinese traditional medicine group had lower expression(P<0.05). There was no statistical difference between two treatment groups(P>0.05).5 RT- PCR result of the expression of MMP-9 and TIMP-1 in each groupThe expression of MMP-9 m RNA in model group, benazepril group and Chinese traditional medicine group was decreased significantly than control group(P<0.05). The expression of MMP-9 m RNA in benazepril group and Chinese traditional medicine group obviously was lower than model group(P<0.05). There was no statistical difference between two treatment groups(P>0.05).The expression of TIMP-1 m RNA in model group, benazepril group and Chinese traditional medicine group was increased significantly than control group(P<0.05). The expression of TIMP-1 m RNA in benazepril group and Chinese traditional medicine group obviously was higher than model group(P<0.05). There was no statistical difference between two treatment groups(P>0.05).Conclusions:1 Yiqi-Yangyin-Qingre-Lishi Recipe could reduce 24-hour proteinuria of rats with Ig A Nephropathy significantly, showing equivalent clinical effects with benazepril.2 Yiqi-Yangyin-Qingre-Lishi Recipe could reduce the pathological changes of kidney tissues to delay the progress of Ig AN.3 Yiqi-Yangyin-Qingre-Lishi Recipe could increase the expression of MMP-9 and reduce the expression of TIMP-1 significantly. These results suggested its protective effect on Ig AN may be related to correcting inbalance of MMP-9 and TIMP-1. |
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