【作者】 汪颖；

【导师】 侯加法；

【作者基本信息】 南京农业大学 ， 临床兽医学， 2009， 硕士

【摘要】 肾代偿在小动物临床上很常见,单侧输尿管阻塞或肾摘除后均可引起对侧肾功能代偿。据文献报道,肾代偿由两种形式——肥大和再生完成。肥大是指肾组织细胞的体积增加,再生是指代偿时肾组织细胞数目增多。有学者认为肾在代偿过程中伴有肾实质进行性损害和细胞凋亡,这与以往关于代偿肾只有代偿性增生和肾血流量增加的观点有较大出入。为此,本试验建立犬肾代偿模型,通过B超检查、尿蛋白分析及显微观察等,研究其代偿肾构型、肾功能及病理组织学变化规律,为临床代偿肾研究提供理论依据。试验Ⅰ、犬代偿肾构型变化的研究目的：用B型超声检测犬代偿肾构型变化。方法：选取13只成年健康犬,对其进行左侧输尿管结扎。分别于结扎前一天和结扎后第1、2、3、4、6、8、10、12、14、17、19及21天测量代偿肾(右肾)肾长、肾宽和肾厚。结果：犬代偿肾第1-21天肾长、肾宽、肾厚与结扎前比较均无显著差异(P>0.05)。结论：代偿肾在一侧输尿管结扎后短期内未发生构型的显著变化。试验Ⅱ、犬代偿肾功能——尿蛋白的研究目的：通过检测尿蛋白含量及蛋白种类,判断代偿肾功能。方法：选取7只成年健康犬,对其进行左侧输尿管结扎后每天采集尿夜,测定34 d内尿蛋白含量,并利用SDS-PAGE电泳方法按蛋白质的分子量大小分离尿蛋白。结果：定量检测尿蛋白发现,输尿管结扎后每天尿中均含有蛋白,且总体呈上升趋势。第28天尿总蛋白含量上升至0.94 mg·mL-1,与结扎前比较差异显著(P<0.05),到第31天其含量达1.9mg·mL-1,与结扎前比较差异极显著(P<0.01)；尿蛋白电泳分离表明,前12天各泳道蛋白条带不明显；之后每天可见明显的蛋白条带,从第13-31天蛋白条带种类逐渐增多,颜色逐渐加深,均见>66.2 KD的条带和<66.2 KD的条带,证明为混合性蛋白尿,提示代偿肾肾小球通透性增强,肾小管重吸收能力下降。第31天后>66.2 KD的条带种类减少。结论：单侧输尿管结扎后12d内尿蛋白含量在正常范围内,SDS-PAGE电泳未分离出尿蛋白。第13-38天出现混合性蛋白尿,提示肾小球及肾小管受损,肾功能降低。试验Ⅲ、犬代偿肾组织病理学研究目的：研究犬代偿肾在代偿过程组织病理学变化。方法：选取7只健康成年犬,分别于左侧输尿管结扎后的第3、7、14、21、28、35及75天采集代偿肾(右肾)组织,样品置于4%多聚甲醛中固定,4℃保存,并对其进行常规组织病理学检查。结果：经HE染色后观察,左侧输尿管结扎后第3天,其代偿肾(右肾)肾小球血管袢充血,肾小球体积稍增大,系膜细胞增生。个别肾小管上皮浊肿变性,部分有脱落；第7天,肾小球血管袢充血、体积增大,肾小囊囊腔变小,系膜细胞增生。个别肾小管轻度浊肿变性；第14天,肾小球充血,体积增大,血管球略有分叶,系膜细胞增生。肾小管上皮水肿,部分变性脱落。部分小管腔内见脱落的上皮细胞。肾小管内有大量蛋白管型；第21天,肾小球体积增大,系膜细胞增加,有中度的增生。肾小管上皮细胞水肿。有些上皮细胞变性、凋亡,管腔内可见脱落的细胞碎片；个别肾小管严重变性,基底膜裸露。管腔内有微量蛋白管型；第28天,肾小球体积较小,系膜细胞轻度增生。’肾小管上皮细胞有疏松变性,部分肾小管上皮细胞脱落至管腔；第35天,肾小球体积较小,系膜细胞轻度增生。肾小管上皮细胞变性脱落。管腔内有微量蛋白管型；第75天,肾小球体积较小,系膜细胞增生。肾小管扩张,肾间质水肿明显,肾小管上皮细胞变性,管腔内有蛋白管型。结论：通过组织病理学检查发现,单侧输尿管结扎后对侧代偿肾在一定时期内有损伤。短期内代偿肾以肾小球体积增大、肾小球充血为主。而系膜细胞增生、肾小管上皮细胞变性、凋亡等病理变化则伴随整个肾代偿的发展过程直至第75天。更多还原

【Abstract】 Canine renal compensation which is induced either by obstruction of the ureter or by removal of the kidney is very common in small animal clinic. According to the report, there are two types of renal compensatory growth process including hypertrophy which means the enlargement of the cell and regeneration which means the increase of the cell quantity that both affected by animal age. Meanwile, some researchers found that the renal parenchyma was damaged and the apoptosis was present during the compensatory period, the view of which is contradictory with others. The purpose of this experiment therefore is to establish the renal compensation model and investigate the configuration, renal function and histopathological change with B-mode ultrasonography, anaslysis of urinary protein and microscopic examination of renal tissue so that to provide for theriocal evidence of compensatory renal research in small animal clinics.ExperimentⅠ:Evaluation of the configuration of compensatory kidney in dogobjective:to evaluate the configuration of canine renal compensation with B-mode ultrasonography. Methods:chosing 13 healthy adult dogs, the left ureters were ligated each dog and the renal length、renal width、and renal depth were measured on compensatory kidney(the right kidney) one day before and 1,2,3,4,6,8,10,12,14,17,19 and 21 days after ureter ligation, respectively. Results:renal length, renal width and renal depth values of compensatory kidney from 1-21 days after ligation were not significantly different compared to that before ligation(P>0.05). Conclusion:the configuration variety of compensatory kidney did not occurr within the short period after ligation. ExperimentⅡ:Evaluation of the function, urinary protein of compensatory kidney in dogObjective:to estimate the function of compensatory kidney by detecting the quantity of the protein and distinguishing the urinary protein origin. Methods:chosing 7 healthy adult dogs, the urine was collected every day after the left ureter was ligated. The urinary total protein were measured within 34 days. and the urinary protein was separated by electrophoresis. Results:results showed that urinary protein could be detected every day after the left ureter was ligated and trended to increase in the total experimental process. The total protein of urine increased by 0.94 mg·mL-1 28 days after the ligation which was significantly different compared with that before ligation (P<0.05) and rased to 1.9 mg·mL-1 31 days after the ligation which was extremely significantly different compared with that before ligation (P<0.01); results of protien electrophoresis showed that the strap on the gelation was not clear on the first 12 days and could be seen obviously on the gelation after that. The types of protein straps were much more than before and the colour was deepened. The strap that both below and up than 66.2 KD appeared from the day 13 to the day 31 which proved that the type of urinary protein belong to miscellaneous proteinuria. It indicated that the glomerular permeability increased and the tubular reabsorption ability decreased; the variety of big relative molecular weight protein was less after 31 days. Conclusion:the urinary protein quantity was in normal range and the protein didn’t appear on gelation by SDS-PAGE within 12 days after ligation. The miscellaneous proteinuria appeared in gelation, indicating that both the glomeruli and the tubule are damaged and the renal function was reduced from day 13 to day 38 after the ligation.ExperimentⅢ:Histopathological changes during the renal compensatory process in dogObjective:to evaluate histopathological changes during the renal comensatory process in dog. Methods:chosing 7 healthy adult dogs, the tissue was collected from compensatory kidney(right kidney) 3,7,14,21,28,35 and 75 days after ligation, respectively. Preserving the kidney tissue sample in 4% paraform with the temperature of 4℃used for HE staining and histopathological examination. Results:according to the hematoxylin and eosin stained slices observation,3 days after the left ureter was ligated, the compensatory renal glomeruli was congested which led to mildly increase of glomeruli volume, besides, the number of mesangial cell was increased. Several epithelial cells of tubule were muddy and swollen and degenerated, part of the cells sloughed off; 7 days after the left ureter was ligated, the capillaries was congested. The Bowman’s capsule was almost "closed" as a result of the increase of the glomeruli volume. The number of mesangial cell was increased. Several epithelial cells of tubule were muddy and swollen mildly; 14 days after the left ureter was ligated, the glomeruli was congested and the volume was increased. The glomeruli was lobulated mildly and the number of mesangial cell was increased. The epithelium cells of tubule were edematous and parts of them were degenerated and sloughed off. Some epithelial cells can be seen in lumen. There were a lot of protein casts in the tubular lumen.21 days after the left ureter was ligated, the volume of the glomeruli was increased and the number of mesangial cell was increased. There were midium hyperplasia of glomeruli. The epithelium cells of tubule were edematous. Some of them were degenerated and apoptotic and sloughed off into the lumen. Several tubules were seriously degenerated and the basement membranes were naked. There were tiny protein casts in the lumen.28 days after the left ureter was ligated, the volume of the glomeruli was a little small and the number of mesangial cell was increased mildly. The epithelium cells of tubule were curmbly degenerated and part of them sloughed off into the lumen; 35 days after the left ureter was ligated, the volume of the glomeruli was a little small and the number of mesangial cell was increased mildly. The epithelium cells of tubule were degenerated and sloughted off. There were tiny protein casts in the lumen; 75 days after the left ureter was ligated, the volume of the glomeruli was a little small and the number of mesangial cell was increased mildly. The tubular lumen was extended. The interstitial edema was abvious. The epithelium cells of tubule were degenerated. There were many protein casts in lumen. Conclusion:the compensatory kidney was found injured by histopathology after the left ureter was ligated. Firstly, the compensatory kidney was characterized by the increase of the glomeruli volume and the congestion of the glomeruli a short time after the left ureter was ligated. Secondly, the pathological change of mesangial cell hyperplasia and the tubular epithelium cell degeneration and apoptosis accompanied with the whole compensatory process 75 days after the left ureter was ligated.更多还原