To Observe The Effects Of Qi Supplementing, Yin Nourishing, Clearing Heat And Expelling Damp Recipe To The Expression Of KLK1 And BK In IgA Nephropathy Rat Kidney

Subject: Ig A nephropathy(Ig AN) is also called Berger disease. It is a primary glomerular diseases which is refers to the deposition of immune complexes in Ig A or Ig A mainly in the mesangium and the capillary loop and the main clinical features is hematuria with or without hematuria occur repeatedly. It is one of the main factors lead to end-stage renal failure. Until now, the pathogenesis of Ig A nephropathy are still not entirely clear. There is no special effective treatment of Western medicine at present. The traditional Chinese medicine has some advantages in the treatment of this disease. Combined with the theory of traditional Chinese medicine, Professor Guo Dengzhou [1-3] and a number of scholars[4-5] through the clinical study of Ig AN patients points out that traditional Chinese medicine can improve the clinical symptoms and laboratory indexes in Ig AN patients. The traditional Chinese medicine has a certain role in the treatment of Ig AN, and achieved satisfactory clinical curative effect. The purpose of this experiment we designed is to investigate the effects of qi supplementing,yin nourishing,clearing heat and expelling damp recipe of Ig A nephropathy rat release kallikrein 1(KLK1) and bradykinin(BK) on the expression of its possible mechanism.Methods: 40 SPF male Sprague-Dauley rats of weight 140 g to 160 g were fed adaptively for 1 week, where the temperature was 20℃ to 24℃ and the Relative humidity was 40% to 60%. Based on the random weight into 4 groups(normal control group, traditional Chinese medicine group, benazepril group and model group)which late seventh day test both urine protein and occult blood were negative. Each group has 10 rats. Using bovine serum albumin(BSA) + lipopoly-saccharide(LPS) + carbon tetrachloride(CCl4) method [6]to copy Ig AN model.Programs are as follows: The rats in traditional Chinese medicine group, benazepril group and model group were treated with BSA by gavage at the dosage of 400mg/kg every other day for 6 weeks; They were injected carbon tetrachloride 0.1ml and castor oil 0.5ml by subcutaneous injection once a week for 9 weeks; On the sixth and eighth weekend, the rats were given 0.05 mg of lipopolysaccharide by tail vein injection. The rats in normal control group, were gave distilled water by gavage in 4ml/kg calculation every other day for 6 weeks; They were injected physiological saline 0.6ml by subcutaneous injection once a week for 9 weeks; The rats were injected physiological saline 0.2 ml by tail vein injection on the sixth and eighth weekend. When the model was established successfully, traditional Chinese medicine group was treated qi supplementing, yin nourishing, clearing heat and expelling damp by gavage. Benazepril group were gave benazepril hydrochloride by gavage. Both normal control group and model group were gave distilled water by gavage at the same dose every day for 8 weeks. We detected 24-hour urinary protein quantitative before modeling, seventh, ninth, thirteenth and seventeenth weekend. All the rats fasting 12 h, weighing, in accordance with the amount of 3.50ml/kg 10% chloral hydrate intraperitoneal injection, anesthesia, in the abdominal aorta blood, serum creatinine(Scr), blood urea nitrogen(BUN) level on the 18 weekend. Kidneys were performed, for HE, Masson, PAS, PASM staining, immune fluorescence method detection of Ig A immune complex deposition in the kidney of rats,reverse transcription polymerase chain reaction(RT-CR) was detected in the rat kallikrein 1(KLK1) and bradykinin(BK) expression, immunohistochemical determination of kallikrein 1(KLK1) expression.Results:1 24-hour urinary protein quantitative: Compared with the normal control group, the last three group rats 24-hour urinary protein quantitative increased significantly(P<0.05).Compared with the traditional Chinese medicine group and benazepril group, model group rats 24-hour urinary protein quantitative increased significantly(P<0.05). Traditional Chinese medicine group and benazepril group rats 24-hour urinary protein quantitative has no significant difference(P>0.05).2 Serum Scr、BUN: No significant difference was found between the normal control group, traditional Chinese medicine group, benazepril group, the model group for rats serum Scr、BUN(P>0.05).3 Observed under light microscope. The normal control group glomerular structural integrity, mesangial and mesangial matrix were no obvious abnormal changes. In the model group, the mesangial cell proliferation and mesangial matrix increase was 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.4 Ig A immunofluorescence:The normal control group rats glomerular mesangial had no immune complex deposition. In the model group we observed the fluorescent green immune complex deposition. The two treatment groups(benazepril group and traditional Chinese medicine group)also observed the fluorescent green immune complex deposition.The two treatment groups compared with the model group, but to a lesser degree.5 The positive expression of KLK1: Compared with the normal control group, the expression of KLK1 in raditional Chinese medicine group, benazepril group and model group was significantly increased(P<0.05=. Compared with the model group, the expression of KLK1 was significantly increased in raditional Chinese medicine group and benazepril group(P<0.05). The expression of KLK1 was no significant difference between the traditional Chinese medicine group and benazepril group(P>0.05).6 The positive expression of BK:Compared with the normal control group, the expression of BK in raditional Chinese medicine group, benazepril group and model group was significantly increased(P<0.05). Compared with the model group, the expression of BK was significantly increased in raditional Chinese medicine group and benazepril group(P<0.05). The expression of BK was no significant difference between traditional Chinese medicine group and benazepril group(P>0.05).Conclusions:1 BSA + LPS + CCl4 can induce Ig AN rat model successfully.2 Qi supplementing,yin nourishing, clearing heat and expelling damp recipe can reduce Ig A nephropathy rats 24-hour urinary protein quantitative and urine red blood cell count, accordingly protect renal function.3 Qi supplementing, yin nourishing, clearing heat and expelling damp recipe can increase the expression of rat kidney KLK1 and BK, thus delay Ig A nephropathy development.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.