摘要
目的探讨细胞因子网络失衡在马兜铃酸肾病(AAN)肾纤维化发生发展中的作用。方法雄性Wistar大鼠46只,SPF级,随机分为对照组(20只)和模型组(26只),模型组用含20mg/(kg.d)马兜铃酸(AA)的关木通浸膏灌胃,对照组用同体蒸馏水灌胃。于第4、8、12周处死动物,取肾组织行HE、PAS、Masson染色,病理学观察分析肾小管损伤和间质纤维化程度,免疫组化分析PCNA、VEGF、TGF-β1蛋白表达,RT-PCR分析VEGF、ET-1、BMP-7mRNA的表达。结果模型组大鼠在第4周即可见明显的肾小管间质损伤,并随时间延长而进行性加重,第12周小管间质形态结构紊乱严重,纤维化程度达31.36%;第4周时肾组织PCNA表达明显增高,第8周时呈下降趋势,第12周时在基底膜裸露的肾小管很少有PCNA阳性表达;在第4周TGF-β1、VEGF表达明显增高,随时间延长TGF-β1呈持续高表达,但VEGF阳性表达逐渐下降。RT-PCR检测发现模型组从第4周起VEGF、ET-1mRNA表达明显升高,第8、12周略有下降,但仍维持在较高水平,而BMP-7随着病变进展表达逐渐减少,第12周达最低水平。结论肾小管严重损伤及快速进展的早期纤维化是AAN的特征之一;肾小管上皮细胞的再生修复能力严重受损,促修复和抑纤维化因子表达不足,而致纤维化因子高表达,可能是AAN纤维化进展的重要原因。
Objective To study the tissue pathology and the expression of related factors during renal tubular-interstitial fibrosis in aristolochic acid （AA） nephropathy in rat. Methods 46 male Wistar rats were randomly divided into 2 groups. The test group consisted of 26 rats which were gavaged with the extract of Caulis Aristolochiae Manshuriensis （CAM） （AA 20mg · kg^-1 · d^-1 ）; the control group consisted of 20 rats which were given with equal volume of potable water. At the end of 4th, 8th, 12th week, the kidneys of each rat were separately harvested. The HE, PAS and Masson staining were used to analyze the degree of tubular damage and interstitial fibrosis, and immunohistochemical method was applied to assess the protein expression of proliferating cell nuclear antigen （PCNA）, vascular endothelial growth factor （VEGF）, transforming growth factor-β1 （TGF-β1） in the renal specimens. The mRNA expression of VEGF, endothelin-1 （ET-1）, bone morphogenetic protein-7 （BMP-7） in renal tissue were determined by RT-PCR respectively. Results A severe renal tubular interstitial damage and an early fibrosis were observed at the end of 12th week, and the interstitial fibrotic area was 31.36%. The protein expression of PCNA was increased at 4th week, but down-regulated after 8th week; the expression of TGF-β1 and VEGF was increased at 4th week, while TGF-β1 was maintatined on a high level with passage of time, but VEGF decreased gradually. The mRNA expression of VEGF and ET-1 increased notably at 4th week, slightly decreased after 8th week, but maintained at a high level. The BMP-7 declined slowly with the progression of pathological changes, reaching its lowest level at 12th week. Conclusion The mechanism of the rapid progression of fibrosis in AAN might be the renal result of severe impairment of regeneration of epithelial cells, lowering of expression of factors of promoting repair and inhibiting fibrosis, while the expression of factors of promoting fibrosis was maintained at a highlevel.
出处
《解放军医学杂志》
CAS
CSCD
北大核心
2006年第12期1141-1145,共5页
Medical Journal of Chinese People's Liberation Army
基金
上海市青年科技启明星基金资助项目(04QMX1453)
上海市自然科学基金资助项目(03ZR14031)
关键词
马兜铃酸
纤维化
肾间质
模型
动物
aristolochic acid
fibrosis
renal interstitium
model, animal
