虫草菌丝粉对慢性马兜铃酸肾损害的防治作用实验研究

【作者】 王冠明；

【导师】 涂晋文；

【作者基本信息】 湖北中医学院 ， 中医内科学， 2006， 博士

【摘要】 本课题从尿液生化学，肾脏病理组织学两个方面对马兜铃酸肾病进行研究，旨在探讨马兜铃酸肾病的发病机理及其防治。 尿液生化学的研究，采用关木通煎剂不间断灌胃及马兜铃酸纯品皮下注射法制作慢性马兜铃酸肾病（CAAN）大鼠模型，随机分为四组：正常组、木通组、治疗组、AA组，给药时间为八周。以大鼠体重增长率，尿NAG酶，24小时尿蛋白定量，尿β₂-微球蛋白（β₂-MG）为观察指标。结果表明，八周后，木通组、治疗组、AA组与正常组相比，大鼠体重明显降低（P＜0.01），24小时尿蛋白定量增加（P＜0.05，P＜0.01），尿NAG酶明显增加（P＜0.01），尿β₂-微球蛋白（β₂-MG）明显增加（P＜0.01）。提示关木通煎剂及马兜铃酸纯品均可导致肾小管功能损伤，而中草药关木通所引起的肾损害与其所含主要成分之一马兜铃酸有密切关系。治疗组与木通组相比，体重降低，24小时尿蛋白定量及尿β₂-微球蛋白（β₂-MG）定量降低，但两者无统计学意义（P＞0.05）；治疗组对降低尿NAG酶明显优于木通组及AA组（P＜0.01）提示要从统计学上肯定人工虫草菌丝粉缓解慢性马兜铃酸肾病所引起的肾损害的作用，还需更长时间的实验室观察。 病理组织学研究，组织取材于尿液生化学研究部分；分组方法相同；采用肾小管间质病理积分，转化生长因子β₁（Transforming growth factor β₁，TGF-β₁），金属蛋白酶抑制剂1（Tissue inhibitor ofmetalloproteinase-1，TIMP-1），基质金属蛋白酶9（Matrix metalloproteinase-9，MMP-9）为观察指标。结果表明，治疗组、木通组及AA组肾小管间质病理积分显著高于正常组（P＜0.01）；其中马兜铃酸纯品组肾小管间质病理积分明显高于木通组（P＜0.05），治疗组积分虽低于木通组，但两者无统计学意义（P＞0.05）。三组TGF-β₁，TIMP-1，MMP-9表达较正常组明显上调（P＜0.01）。提示①小剂量AA反复作用于肾小管上皮细胞，可使该细胞活化，活化的肾小管上皮细胞可释放转化生长因子β₁（TGF-β₁）因子，通过细胞间的“串话”（cross talking）作用于肾间质成纤维细胞，激活后者分泌细胞外基质（ECM），导致肾间质纤维化；分化成肌成纤维细胞的肾小管上皮细胞，还能直接分泌ECM，加重肾间质纤维化。②AA可刺激肾小管上皮细胞MMP-9的表达，导致ECM合成增加，同时激活其降解作用，TIMP-1可能因反馈调节而表达增强，最终引起MMP-9／TIMP-1失平衡，抑制胶原降解，ECM积聚，加重肾间质纤维化。这两种机制均可能参与了慢性AAN的形成。其中木通组TGF-β₁，TIMP-1表达较治疗组上调（P＜0.01），MMP-9表达较治疗组下调（P＜0.01），提示人工虫草菌丝粉可抑制更多还原

【Abstract】 To establish a model of chronic aristolochic acid nephropathy （CAAN） in rats and to investigate the pathogenesis of its renal interstitial fibrosis include alleviate its renal injury by Chinense Caterpillar fungus mycleium powderIn the biochemical researches, Wistar rats were distributed into fourgroups. Groupl was normal control ,Group 2 and 4 were model control, 3 treated group. Group 2 was received Aristolochia manshuriensis Kom. decoction 10g/kg body weight ,Group 3 was received Aristolochia manshuriensis Kom. decoction lOg/kg and Chinense Caterpillar Fungus mycelium powder 5g/kg body weight once daily for 8 weeks. Group 4 was subcutaneous injected aristolochic acid lOmg/kg body weight once daily for 8 weeks . After administration ,urine samples were obtained for NAG, β₂-MG and protein evaluation.It was found that body weights in group 2 、 3and group 4 were decreased compare with group 1. Protein of 24 hours urine, NAG and β₂-MG of the two groups were higher than groupl （ P<0. 05, P<0.01 ） . There for , the results of study suggest that Aristolochia manshuriensis Kom.decoct ion and aristolochic acid could result in tubulopathy. We also found that rats of Chinense Caterpillar Fungus mycelium powder treated resulted in improved body weights, Protein of 24 hours urine, NAG and β₂-MG at eight weeks point compared with Group 2 that rats were received Aristolochia manshuriensis Kom. decoction 10g/kg. There for , the results of study suggest that we need more times to make model to confirm the function of Chinense Caterpillar Fungus mycelium powder on alleviating the renal injury of chronic aristolochic acid nephropathy （CAAN）.In the histological researches , tubulointerstitia1 histological points. Transforming growth factor β₁ （TGF-β ₁）, Tissue inhibitor of metalloproteinase-1 （TIMP-1） and Matrix metallo proteinase-9 （MMP-9） were determined.It was found that in the group 2、 3and group 4 the tubulo -interstitial histological points increased compared with those of the normal control （P<0. 01 ） , and the points of group 4 was the highest, while the group 3 had the lowest ones . the更多还原