血管生成素及其受体在腹膜透析腹膜组织中的表达

发布时间：2018-05-12 02:11

本文选题：腹膜透析 + 血管新生　；参考：《南昌大学》2011年硕士论文

【摘要】：目的：众所周知,长期的腹膜透析会导致腹膜功能逐渐出现衰退,最终导致超滤衰竭,使得尿毒症患者最终不得已退出腹膜透析。新近研究发现,腹膜发生血管新生是患者腹膜功能衰退的主要原因之一。本实验旨在通过建立尿毒症腹膜透析大鼠模型,了解其腹膜组织血管生成素及其受体的表达情况以及引起血管生成素及其受体变化的影响因素,以探讨腹膜新生血管的发生机制。方法：选取健康SD雄性大鼠30只,体重约为180-220g,均购于江西省中医院医学动物实验中心。适应性喂养一周以后,随机选取其中22只大鼠建立经典的5/6肾切除术尿毒症模型,造模四周后采集大鼠尾静脉测血肌酐、尿素氮值,肾切除术后4周,造模成功的大鼠腹腔内予植入自制的腹透管,随机分为尿毒症组(NS组)及腹透组(PD组)。尿毒症组每日给予20ml生理盐水腹腔灌注,腹透组则每日给予20ml 4.25%百特腹透液腹腔灌注,共持续六周。余下未造模的8只大鼠拟为对照组(C组),行假手术且不予任何液体灌注。所有的大鼠均正常饮食饮水,灌液六周后取大鼠腹膜组织,观察腹膜血管新生情况；免疫组织化学染色及逆转录聚合酶链式反应技术(reverse transcription-polymerise chain reaction, RT-PCR)检测腹膜组织血管生成素及其受体的表达变化。结果：(1)实验表明PD组相对其他两组新生血管数显著增多,差异具有统计学意义(P0.05);NS组新生血管数较C组为多,差异具有统计学意义(P0.05)。(2)免疫组化显示各组大鼠血管生成素1(Angpt-1)阳性细胞数比较P0.05,无统计学意义；血管生成素2(Angpt-2)及其受体2(Tie-2)阳性细胞数PD组相对于C组和NS组显著增加,差异具有统计学意义(P0.05)；NS组较C组亦有增加,差异具有统计学意义(P0.05)。(3)RT-PCR显示Angpt-1mRNA条带半定量分析结果在三组间无统计学意义(P0.05)；PD组Angpt-2 mRNA和Tie-2mRNA条带深于其他两组,半定量分析差异具有统计学意义(P0.05);NS组Angpt-2 mRNA和Tie-2 mRNA半定量分析结果高于C组,差异具有统计学意义(P0.05)。(4) Angpt-2阳性细胞数与腹膜新生血管数之间存在直线正相关关系(P0.05, R0.855); Angpt-2mRNA半定量分析结果与腹膜新生血管数之间亦呈直线正相关关系(P0.05,R0.742)。结论：尿毒症状态及腹膜透析液均会上调腹膜血管生成素-2及其受体的表达,但未见血管生成素1的表达出现明显变化。腹膜血管生成素-2及其受体的表达与腹膜血管新生密切相关。
[Abstract]:Objective: it is well known that long-term peritoneal dialysis can lead to gradual decline of peritoneal function and eventually to ultrafiltration failure, which makes uremic patients finally have to withdraw from peritoneal dialysis. Recent studies have found that peritoneal angiogenesis is one of the main causes of peritoneal dysfunction in patients. The purpose of this study was to investigate the expression of angiopoietin and its receptors in peritoneal tissues and the factors affecting the changes of angiopoietins and their receptors by establishing a rat model of peritoneal dialysis with uremia. To explore the mechanism of peritoneal neovascularization. Methods: 30 healthy male SD rats, weighing 180-220 g, were purchased from the Medical Animal experiment Center of Jiangxi traditional Chinese Medicine Hospital. After one week of adaptive feeding, 22 of them were randomly selected to establish the classic 5 / 6 nephrectomy uremic model. The blood creatinine, urea nitrogen and blood urea nitrogen were collected from the tail vein of the rats for 4 weeks after nephrectomy. The successful rats were implanted into abdominal cavity and randomly divided into uremia group (NS group) and peritoneal dialysis group (PD group). The uremic group was intraperitoneally perfused with 20ml saline and the peritoneal dialysis group with 20ml 4.25 hundred per day for six weeks. The remaining 8 rats were selected as control group C, sham-operated without any fluid infusion. All the rats were fed and drinking normal diet. After 6 weeks of perfusion, the peritoneal tissues were taken out to observe the situation of peritoneal angiogenesis. Immunohistochemical staining and reverse transcription-polymerise chain reaction (RT-PCR) were used to detect the expression of angiopoietin and its receptor in peritoneal tissues. Results compared with the other two groups, the number of neovascularization in PD group was significantly higher than that in the other two groups, and the difference was statistically significant (P 0.05) the number of neovascularization in NS group was higher than that in C group. The number of Angiopoietin-1Angpt-1) positive cells in each group was not significantly higher than that of P0.05, and the number of Angiopoietin-2Angpt-2) and its receptor 2tie-2) in PD group was significantly higher than that in C group and NS group. The results of semi-quantitative analysis of Angpt-1mRNA bands showed that there was no significant difference between the three groups in the Angpt-2 mRNA and Tie-2mRNA bands in the PD group, and the Angpt-2 mRNA and Tie-2mRNA bands in the PD group were deeper than those in the other two groups. The difference of semi-quantitative analysis was statistically significant. The results of Angpt-2 mRNA and Tie-2 mRNA in NS group were higher than those in C group. There was a linear positive correlation between the number of Angpt-2 positive cells and the number of peritoneal neovascularization (P0.05, R0.855), and there was also a linear positive correlation between the semi-quantitative analysis of Angpt-2mRNA and the number of peritoneal neovascularization. Conclusion: both uremia and peritoneal dialysate can up-regulate the expression of angiopoietin-2 and its receptor, but there is no obvious change in the expression of angiopoietin-1. The expression of angiopoietin-2 and its receptor is closely related to peritoneal angiogenesis.
【学位授予单位】：南昌大学
【学位级别】：硕士
【学位授予年份】：2011
【分类号】：R692.5;R-332

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