| Objective: This experiment is designed to study the mechanism of qi supplementing yin nourishing clearing heat and expelling damp recipe to Ig A nephropathy rat kidney through observing 24-hour urinary protein quantitative, renal function,using immunohistochemical method, RT-PCR determine method to detection the expression of NF-κB and MCP-1 in renal tissue. The aim is to provide new ideas and methods for clinical treatment of Ig A nephropathy.Methods: 40 SPF male Sprague-Dauley rats of 6 week old and 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 which late seventh day test both urine protein and occult blood were negative:10 in normal control group, 10 in model group, 10 in benazepril group, 10 in qi supplementing yin nourishing clearing heat and expelling damp group. The rats in 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. The rats in last three groups were treated with BSA(concentration:100 g/L)by gavage at the dosage of 400mg/kg every other day for 6 weeks; They were injected carbon tetrachloride 0.1 ml and castor oil 0.5 ml by subcutaneous injection once a week for 9 weeks; On the sixth and eighth weekend, the rats were given 0.05 mg of lipopolysaccharide(concentration: 0.25g/L)by tail vein injection. From the 10 th week, qi supplementing yin nourishing clearing heat and expelling damp group were gave 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 h urine protein before modeling, in the 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. To observe the pathological changes of renal tissue in rats. Using immunohistochemical method, RT-PCR determine method to detection the expression of NF-κB and MCP-1 in renal tissue.Results:1 24 h urine protein quantitative24h urine protein quantitative: Medication intervention, in the seventeenth week, the two treatment groups(benazepril group and qi supplementing yin nourishing clearing heat and expelling damp group) was no significant difference(P>0.05). Two treatment group compared with the model group, the urine protein quantitative decreased obviously, there was significant difference(P<0.05).2 Renal functionThe Scr and BUN were no significant differences in each group(P>0.05).3 Renal pathologic changesThe results of optical microscope: The normal control group rats glomerular structured integrity, mesangial and matrix had no obvious changes; In the model group, the mesangial cell proliferation and mesangial matrix increase was observed.In the two treatment groups(benazepril group and qi supplementing yin nourishing clearing heat and expelling damp group), the mesangial cell proliferation and mesangial matrix increased, but compared with the model group, pathological changes was reduced.Immunofluorescence observation : Observation of immune complex deposition there was no deposition in glomerular mesangial region. 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 qi supplementing yin nourishing clearing heat and expelling damp group)also observed the fluorescent green immune complex deposition.The two treatment groups compared with the model group, but to a lesser degree.4 Immunohistochemical method to detection the expression of NF-κB and MCP-1 in renal tissueThe positive expression of NF-κB:The normal control group is weak, the other three group were significantly enhanced(P<0.05);The two treatment groups(benazepril group and qi supplementing yin nourishing clearing heat and expelling damp group) expression decreased than the model group(P<0.05). The two treatment groups were no significant differences(P>0.05).The positive expression of MCP-1:The normal control group is weak, the other three group were significantly enhanced(P<0.05);The two treatment groups(benazepril group and qi supplementing yin nourishing clearing heat and expelling damp group) expression decreased than the model group(P<0.05). The two treatment groups were no significant differences(P>0.05).We abserved that NF-κB and MCP-1 expression in renal tissue was positively related to by immunohistochemical method.5 RT-PCR determine method to detection the expression of NF-κB and MCP-1 in renal tissueBoth NF-κB and MCP-1, the normal control group is weak, the other three group were significantly enhanced(P<0.05);The two treatment groups(benazepril group and qi supplementing yin nourishing clearing heat and expelling damp group) expression decreased than the model group(P<0.05). The two treatment groups were no significant differences(P>0.05).We abserved that NF-κB and MCP-1 expression in renal tissue was positively related to by RT-PCR determine method.Conclusion:1 Qi supplementing yin nourishing clearing heat and expelling damp recipe can decrease urine protein in Ig A nephropathy rats, thus protecting kidney function.2 Qi supplementing yin nourishing clearing heat and expelling damp recipe can improve the pathological tissue injury of renal tissue of Ig A nephropathy rats, and delay the progression of Ig A nephropathy.3 Qi supplementing yin nourishing clearing heat and expelling damp recipe prevent the proliferation, mesangial sclerosis, mesangial matrix deposition, delaying the progression of Ig A nephropathy maybe through the reduced expression of NF-κB and MCP-1 in renal tissue of Ig A nephropathy rats, and reduce inflammation. |
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