| Objective: Ig A nephropathy(Ig AN) is a common primary glomerular disease, with renal pathology showed immune complexes Ig A mainly in the glomerular mesangial area features. Main manifestations were hematuria, accompanied by varying degrees of proteinuria, hypertension and renal function damaged, leading to end-stage renal disease common primary glomerular disease, 10% ~ 20% of adult patients within 10 years progress to end-stage renal disease. A large number of studies show that TCM has certain advantages in the treatment of this disease. Professor Guo Deng Zhou according to the pathogenesis of "deficiency of both qi and Yin, accompanied by damp heat", the proposed with Supplementing Qi and nourishing yin and clearing heat and removing dampness and clinical achieved good effect [1-2]. Through the study of the protective effect on the kidney of Ig AN rats and its effect on the B1 receptor, B2 receptor, and explore its possible mechanism.Methods: 40 healthy male SD rats, weighing between 140 g and 160 g were fed adaptively at the temperature of 20℃to 24℃, relative humidity 40%- 60% of the environment for 7 days, at the end of the first 7 days, according to the weight they were divided for four groups which late seventh day test both urine protein and occult blood were negative: control group, model group, traditional Chinese medicine group and benazepril group. Reference of the Tang Ying[3] modeling method to model replication: bovine serum albumin with distilled water and prepared into 100g/L concentration, to 4ml/kg(400mg/kg) was administered by gavage every other day for 42 days; carbon tetrachloride 0.1ml+ castor oil 0.5ml subcutaneous injection every other day for 63 days; lipopolysaccharide with deployment of sodium chloride solution, the concentration of 0.25g/L, at the 6th and 8th week tail vein injection of 0.05 mg(0.2ml). The normal group rats were given the same dose of distilled water gavage every other day for 42 days; subcutaneous injection of sodium chloride solution(0.6ml), once a week for 9 weeks; at the 6th and 8th week tail vein injection of sodium chloride solution 0.2ml. When the model was established successfully at the 64 th day were given each corresponding drug intervention. Distilled water in normal group and model group were given corresponding dose gavage for 56 days. The 50 th day and 64 th day, and 92 th day and 120 th day were detected in rats of 24 h urinary protein. At the end of the 18 th week, fasting 12 h, with 3.5ml/kg of 10% chloral hydrate intraperitoneal anesthesia, blood from the femoral artery, the rats were sacrificed, separation of serum, serum creatinine(Scr) and urea nitrogen(BUN) level. Renal tissue samples were collected, stained by HE, Masson, PAS, PASM staining, immunofluorescence detection of Ig A immune complex deposition in the kidney of rats and reverse transcription polymerase chain reaction(RT-PCR) and immunohistochemistry determination of B1 receptors and B2 receptor.Results: 1 Comparison of 24 hour urinary protein quantitative 1.1 Compared with the same period 24 hour urinary protein quantitativeThere was no significant difference in four groups before model establishment(P>0.05).At the 50 th day, compared with control group, 24 hour urinary protein quantitative 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 64 th day, compared with control group, 24 hour urinary protein quantitative 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 92 th day, compared with control group, 24 hour urinary protein quantitative 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 120 th day, compared with control group, 24 hour urinary protein quantitative 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). 1.2 The comparison of same group in different periods of 24 hours urinary protein quantitativeThe comparison in the control group: There was no significant difference in five periods.The comparison in the benazepril group: the 50 th day than before modeling significantly increased, and has statistical difference(P<0.05); the 64 th day than the 50 th day increased significantly, and has significant difference(P<0.05); the 92 th day, the 120 th day compared with the 64 th day were significantly reduced, and has significant difference(P< 0.05); the 120 th day compared with the 92 th day decreased obviously, and has statistical difference(P<0.05).The comparison in the Chinese traditional medicine group: the 50 th day than before modeling significantly increased, and has statistical difference(P<0.05); the 64 th day than the 50 th day increased significantly, and has significant difference(P<0.05); the 92 th day, the 120 th day compared with the 64 th day were significantly reduced, and has significant difference(P<0.05); the 120 th day compared with the 92 th day decreased obviously, and has statistical difference(P<0.05).The comparison in the model group: the 50 th day than before modeling significantly increased, and has statistical difference(P<0.05); the 64 th day than the 50 th day increased significantly, and has significant difference(P<0.05); the 64 th day, the 92 th day and the 120 th day compared, there no significant difference(P>0.05). 2 renal pathological changes under the light microscopeThe control group glomerular structural integrity, glomerular mesangial cells and matrix had no obvious abnormal change; in the model group, the mesangial cell proliferation and mesangial matrix increase were observed. The mesangial cell proliferation and mesangial matrix increased in traditional Chinese medicine group and benazepril group, but compared with the model group, pathological changes was reduced. 3 immunofluorescence of Ig A expression in the kidneyThe control group: glomerular mesangial area showed no obvious immune complex deposition. The model group: glomerular visible mesangial immune complex granular or lump deposit. The benazepril group and the traditional Chinese medicine group showed different degrees of mesangial deposition of immune complexes, but compared with the model group, the degree of immune complexes deposition was lighter than that in the model group. 4 expression of B1 reporter in kidney:Immunohistochemistry: compared with the control group, the expression of B1 reporter in the traditional Chinese medicine group, benazepril group and model group was significantly increased(P<0.05); Compared with the model group, the expression of B1 reporter were significantly decreased in the Chinese medicine group and benazepril group(P<0.05). The expression of B1 reporter was no significant difference between the traditional Chinese medicine group and benazepril group(P>0.05).RT-PCR: compared with the control group, the expression of B1 reporter in the traditional Chinese medicine group, benazepril group and model group was significantly increased(P<0.05); Compared with the model group, the expression of B1 reporter were significantly decreased in the Chinese medicine group and benazepril group(P<0.05). The expression of B1 reporter was no significant difference between the traditional Chinese medicine group and benazepril group(P>0.05). 5 expression of B2 reporter in kidneyImmunohistochemistry: compared with the control group, the expression of B2 reporter in the traditional Chinese medicine group, benazepril group and model group was significantly increased(P< 0.05); Compared with the model group, the expression of B2 reporter were significantly decreased in the Chinese medicine group and benazepril group(P<0.05). The expression of B2 reporter was no significant difference between the traditional Chinese medicine group and benazepril group(P>0.05).RT-PCR: compared with the control group, the expression of B2 reporter in the traditional Chinese medicine group, benazepril group and model group was significantly increased(P< 0.05); Compared with the model group, the expression of B2 reporter were significantly decreased in the Chinese medicine group and benazepril group(P < 0.05). The expression of B2 reporter was no significant difference between the traditional Chinese medicine group and benazepril group(P>0.05).Conclusion:1 Qi supplementing yin nourishing clearing heat and expelling damp recipe can reduce the 24 hours urine protein in Ig A nephropathy rats.2 Qi supplementing yin nourishing clearing heat and expelling damp recipe can reduce the expression of bradykinin B1 receptor, thereby reducing proteinuria and protecting kidney function.3 Qi supplementing yin nourishing clearing heat and expelling damp recipe can raise the expression of bradykinin B2 receptor, thereby reducing proteinuria and protecting kidney function.4 Qi supplementing yin nourishing clearing heat and expelling damp recipe can delay the pathological lesions of Ig A nephropathy in rats, thereby delaying disease progression. |
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