【摘要】 目的探讨铁超负荷与肾间质纤维化(RIF)的关系。方法 2012年2月—2013年1月,选取同批清洁级健康Wister雄性大鼠84只,按体质量分层后随机分为假手术组、模型组、模型+右旋糖酐铁组(右旋糖酐铁500mg/kg)、模型+右旋糖酐铁+去铁敏组(右旋糖酐铁500 mg/kg+去铁敏150 mg·kg-1·d-1)、模型+右旋糖酐铁+N-乙酰半胱氨酸(NAC)组(右旋糖酐铁500 mg/kg+NAC 50 mg·kg-1·d-1)、模型+右旋糖酐铁+维生素E组(右旋糖酐铁500 mg/kg+维生素E 50 mg·kg-1·d-1),每组14只;根据术后观察时间的不同,将每组大鼠采用简单随机抽样法进行均分并分别于术后7 d及术后10 d进行观察。除假手术组外,其余5组均采用单侧输尿管结扎模型。术后第7、10天分别取大鼠左侧肾脏组织制成切片,进行苏木素-伊红(HE)染色、马松染色(Masson染色)及普鲁士兰染色于光镜下观察,比较各组RIF程度和阳性细胞数。结果 HE染色:模型组第7天肾间质有少量炎性细胞,肾小管上皮细胞变矮、肿胀、细胞核变长,出现颗粒和空泡变性;第10天肾间质炎性细胞浸润、充血,肾小管扩张,肾小管上皮细胞萎缩、坏死、部分脱失,基底膜出现断裂、溃解,肾间质出现轻度增宽,肾小球无明显病变。模型+右旋糖酐铁组病变较模型组明显,模型+右旋糖酐铁+去铁敏组、模型+右旋糖酐铁+NAC组、模型+右旋糖酐铁+维生素E组病变较模型组轻,假手术组未见明显变化。Masson染色:第7天模型组肾间质仅有少量胶原纤维,模型+右旋糖酐铁组可见大量胶原纤维增生;第10天模型组可见少量胶原纤维,模型+右旋糖酐铁组可见明显胶原纤维增生、肾间质增宽,扩张的肾小管基底膜旁胶原纤维增生,肾小球周围可见明显胶原纤维,肾小球中无胶原纤维增生,假手术组肾小管及肾间质无明显改变。6组大鼠第7天及第10天RIF程度比较,差异有统计学意义(P<0.05);模型+右旋糖酐铁组RIF程度重于模型组,模型+右旋糖酐铁+去铁敏组、模型+右旋糖酐铁+NAC组、模型+右旋糖酐铁+维生素E组RIF程度轻于模型+右旋糖酐铁组(P<0.05);各组第10天RIF程度重于第7天(P<0.05)。普鲁士兰染色:第7天模型+右旋糖酐铁组肾小管及肾间质内可见较多蓝色颗粒,模型+右旋糖酐铁+维生素E组肾小管及肾间质可见少量蓝色颗粒,其余组未见蓝色颗粒;第10天各组均未见蓝色颗粒。第7天模型+右旋糖酐铁组阳性细胞数高于模型+右旋糖酐铁+维生素E组(t=5.85,P=0.000 1)。结论右旋糖酐铁会加重RIF,但去铁敏和抗氧化剂(NAC和维生素E)可减轻由于铁超负荷导致的RIF;铁超负荷加重RIF可能与其氧化应激的过程有关。更多还原

【Abstract】 Objective To investigate the relationship between iron overload and renal interstitial fibrosis( RIF).Methods A total of 84 clean- grade healthy Wister male rats of the same patch were selected. According to the body mass of the included rats,we divided them into six groups: sham operation group,model group,model + iron dextran group( 500 mg / kg iron dextran),model + iron dextran + DFO group( 500 mg / kg iron dextran + 150 mg·kg- 1·d- 1DFO),model + iron dextran+ NAC group( 500 mg / kg iron dextran + 50 mg·kg- 1·d- 1NAC) and model + iron dextran + vitamin E group( 500 mg / kg iron dextran + 50 mg · kg- 1·d- 1vitamin E). Each group included 14 rats. According to different postoperative observation time,the 14 rats in each group were further divided into 7 d group and 10 d group,with each group 7 rats. Apart from the sham operation group,the rest five groups were all administrated with unilateral ureteral ligation. On day 7 and day 10 after operation,left kidney tissues of the rats were sampled and sliced. And the kidney slices were dyed with HE,Masson and Prussian blue and were observed under light microscope to compare the RIF degree and the number of positive cells. Results HE dyeing: for model group,on day 7,a small amount of renal interstitial inflammatory cells were noted,renal tubular epithelial cells became shortened and swollen,and lengthened cell nucleus, particles and vacuolar degeneration were also observed in renal tubular epithelial cells; on day 10, cellular infiltration and congestion occurred in renal interstitial inflammatory cells, the kidney tubules expanded,cellular atrophy,necrosis and partial attrition were found in renal tubular epithelial cells,the breakage and burst of basilar membrane occurred, slight broadening was noted in renal interstitium, and no obvious lesion occurred in glomerulus and kidney tubules; compared with model group,model + iron dextran group had more serious lesion,and model+ iron dextran + DFO group,model + iron dextran + NAC group and model + iron dextran + vitamin E group had less serious lesion,while no obvious lesion occurred in sham operation group. Masson dyeing: for model group,on day 7, only a small amount of collagenous fiber was noted in renal interstitium,while a large amount of collagenous fiber was noted in model + iron dextran group; on day 10,collagenous fiber was still of a small amount in model group,while obvious hyperplasia of collagenous fiber and the broadening of renal interstitium were observed in model + iron dextran group,hyperplasia of collagenous fiber beside the basilar membrane of expanded kidney tubules was noted, obvious collagenous fiber around glomerulus occurred, no hyperplasia of collagenous fiber occurred within glomerulus,and no obvious change in kidney tubules and renal interstitium was observed in sham operation group; the rats had significantly different( P < 0. 05) RIF degrees between day 7 and day 10; model+ iron dextran group had higher( P < 0. 05) RIF degree than model group,and model + iron dextran + DFO group,model + iron dextran + NAC group and model + iron dextran + vitamin E group had lower( P < 0. 05) RIF degrees than model + iron dextran group; the six groups had higher RIF degrees on day 10 than on day 7. Prussian blue: on day 7,many blue particles were found in kidney tubules and renal interstitium within model + iron dextran group,while only a few blue particles were found in kidney tubules and renal interstitium in model + iron dextran + vitamin E group,and no blue particles were found in the other groups; on day 10,no blue particles were found in any group; on day 7,model + iron dextran group was higher than model + iron dextran+ vitamin E group in the number of positive cells( t = 5. 85,P = 0. 000 1). Conclusion Iron dextran could aggravate the RIF. NAC and Vitamin E could alleviate RIF induced by iron overload. That RIF iron overload could aggravate RIF may be related with the process of oxidative stress.更多还原