| Objective: The basic pathological characteristics of Ig A nephropathy is the appearance of Ig A as the main immune complex depositon in the glomerular mesangial area and the capillary loops, at the same time the major histologic change of Ig AN is glomerular mesingial proliferation which is a kind of primary glomerulonephritis. Ig A nephropathy incidence is high in our country and it is a common cause of end-stage renal disease among glomerular diseases. Yet the specific pathogenesis of this disease is not very clear, no particularly effective drugs have been found for it and the clinical effects were not satisfied. My tutor, Professor Guo dengzhou state, according to his clinical experiences, that the theory of "qiyin-liangxu-shire-bingzhong" and"yiqi-yangyin-qingre-qushi" traditional Chinese medicine to treat Ig A which usually could get significantly clinical efficacy. Our experiment explored the angiotensin-converting enzyme 2（ACE2） and prolycarboxypeptidase（PRCP）expression in Ig AN rats to provide experimental basis and theoretical basis for the treatment of Ig AN.Methods: 40 healthy male SD rats, SPF, were randomly divided into 4groups, the normal group（N group）, model group（M group）, traditional Chinese medicine group（Z group）, and benazepril group（B group）, ten rats in each group. Except for N group, rest groups were copied into Ig A model. The definite means are as follows: BSA was made up by distilled water into100g/L and then the rats received gavage in a dosage of 400mg/kg for 42 days.Meanwhile, 0.5ml castor oil and 0.1ml carbon tetrachloride subcutaneous injection, once seven day for 63 days and at the 42 th and 56 th day the rats were injected by tail vein 0.05 mg lipopolysaccharide. N group was given400mg/kg of distilled water every other day for 42 days, 0.6ml saline subcutaneous injection, once seven days for 63 consecutive days and at the42 th and 56 th day the rats were given 0.2ml sodium chloride solution by tail vein. From the 64 th day, the B group was given benazepril 1.05mg/kg once a day, the Z group was given "yiqi-yangyin-qingre-qushi" traditional Chinese medicine once a day, and the M group was given the same dose distilled water as N group for 56 days. In the process of the experiment, the rats were feeded in metabolic cages and collect 24-hour urine to test 24-hour urine protein quantitative determination on the first day, the fiftieth day, the sixty-fourth day, the ninty-second day and the one hundred and twentieth day. At the end of the experiment the rats were sacrificed. We operated on the rats to get kidney for HE, PAS, Masson, PASM staining and immunofluorescence to observe the renal pathological changes, immunohistochemistry and reverse transcription polymerase chain reaction to confirm the expression of ACE2 and PRCP.Results:1 The effects of "yiqi-yangyin-qingre-qushi" traditional Chinese medicine on 24-hour urinary protein quantitative:On the first day after making model, no significant differences were seen in four groups（P>0.05）.On the 50th day, compared with N group, the 24h-UTP of the other three groups were increased（P<0.05）, however, there was not obvious difference among these three groups（P>0.05）.On the 64th day, compared with N group, the other groups were significantly increased（P<0.05）, no obvious differences appeared（P>0.05）.On the 92th day, M group, Z group and B group still presented a higher level of 24h-UTP than N group（P<0.05）. The 24h-UTP expression in Z group and B group were less than M group（P<0.05）,but there were no differences between in the two groups（P>0.05）.On the 120th day, except for N group, others still kept a high level expression of 24h-UTP（P<0.05）. Compared with M group, Z and B group showed greatly reduced（P<0.05）, no statistically differences were found in the two groups（P>0.05）.2 Renal histopathologyLight microscopy: N group rats’ glomerular structure were intact, there was no obvious changes in the mesangial and matrix. M group appear distinct mesangial cells and matrix proliferation, Z group and B group were less than M group.Immunofluorescence: There were no deposition of immune complex in N group, however, obvious immune complex could been seen in M group. The Z group and B group presented less deposition of immune complex than group.3 To observe the expression of ACE2 and PRCP in rats’ kidney by immunohistochemistry and reverse transcription polymerase chain reaction3.1 To observe the expression of ACE2 and PRCP in rats’ kidney by immunohistochemistry:ACE2 expression in N group was much more than other three groups（P﹤0.05）, the expression of ACE2 in the Z group and B group were higher than M group（P﹤0.05）, but there was no significant difference between Z group and B group（P﹤0.05）.M group presented lower PRCP expression than N group（P﹤0.05）, the expression of PRCP in Z group and B group were much higher than N group（P﹤0.05）, the two groups showed no significant difference（P﹥0.05）.3.2 To observe the m RNA expression of ACE2 and PRCP in rats’ kidney by reverse transcription polymerase chain reactionM group, Z group B group showed significantly lower ACE2 m RNA expression than N group（P﹤0.05）, drug-treated groups appeared higher ACE2 m RNA expression than M group（P ﹤ 0.05）, no obvious difference could be seen in the drug-treated groups（P﹥0.05）.The PCRP m RNA expression in M group was lower than N group（P﹤0.05）, drug-treated groups were higher than N group（P﹤0.05）, but there was no significant difference between Z group and B group（P﹥0.05）.Conclusions:1 "Yiqi-yangyin-qingre-qushi" traditional Chinese medicine could significantly reduce Ig AN rats’ 24-hour urine protein level, and its clinicaleffect was similar to benazepril.2 "Yiqi-yangyin-qingre-qushi" traditional Chinese medicine could reduce renal pathological tissue damage and slow disease progression.3 "Yiqi-yangyin-qingre-qushi" traditional Chinese medicine could increase the expression of ACE2 and PRCP in rats’ kidney which confirmed that its mechanism probably was inhibiting RAS system and promote the KKS system. |
