SHR大鼠肾脏及循环中CD13表达水平与原发性高血压发病关系的实验研究

发布时间：2018-04-08 09:54

  本文选题：原发性高血压　切入点：原发性高血压大鼠　出处：《安徽理工大学》2015年硕士论文

【摘要】：目的：初步探讨CD13分子在原发性高血压疾病中的表达水平及其在高血压发生发展中的作用。 方法：4周龄SPF级雄性自发性高血压大鼠(spontaneously hypertensive rat,SHR)及其野生对照组京都维斯塔尔大鼠(Wistar-Kyoto rat, WKY)各32只,按简单随机法将SHR、WKY大鼠各分为4组,每组8只,分别在4周(O期)、6周(高血压起始期)、10周(高血压成型期)、23周(靶器官损害期)进行实验观察。采用无创尾动脉血压测量仪动态监测SHR、WKY的血压情况；应用ELISA法动态检测上述大鼠的24h尿蛋白情况；Trizol法提取各组大鼠肾脏组织总RNA,并逆转录为cDNA,RT-PCR法动态检测大鼠肾脏中CD13mRNA的表达水平；取各组大鼠肾脏皮质髓质纵切面2mm厚度组织,石蜡法常规脱水包埋连续切片,应用免疫组织化学技术动态检测CD13蛋白的表达水平。应用ELISA法动态检测CD13、AngⅢ、AngⅣ在大鼠血浆中的表达水平并分析上述指标与SHR血压变化的相关性；采用HE及MASSON染色检测各期大鼠肾脏病理改变情况。 结果：SHR大鼠体重在观察期间内均低于同期野生对照WKY大鼠,在6周、10周、23周分别为(160.2±5.3)g vs.(186.8±6.8)g、(230.6±13.5)g vs.(252.3±5.5) g,(310.7±9.1) g vs.(357.4±7.0)g,两组差异均有统计学意义(均为P0.05)。SHR大鼠血压值亦高于同期野生对照WKY大鼠,在6周、10周、23周分别为(128.8±10.3)mmHg vs.(109.1±3.8)mmHg、(152.6±8.6)mmHg vs. mmHg、(203.9±13.2) mmHg vs.(112.9±6.9)mmHg,两组差异均有统计学意义(均为P0.05)。RT-PCR结果显示,SHR肾脏中CD13mRNA表达水平高于同期野生对照WKY大鼠。在6周、10周、23周分别为0.36+0.05vs.0.25+0.03、0.61±0.08vs.0.28±0.03、0.62±0.09vs.0.17±0.02,两组差异均有统计学意义(均为P0.05)。免疫组化染色结果显示,SHR肾脏中CD13蛋白表达水平亦高于同期野生对照WKY大鼠。在6周、10周、23周分别为0.42±0.12vs.0.25±0.15、0.57±0.13vs.0.38±0.09、0.73±0.12vs.0.37±0.19,两组差异均有统计学意义(均为P0.05)。血浆中CD13的表达低于同期野生对照WKY大鼠。在10周、23周分别为(107.4±22.3) ng/L vs.(147.1±25.5) ng/L、(87.04±20.7)ng/L vs.(150.1±31.2)ng/L,两组差异均有统计学意义(均为P0.05)。此外,SHR大鼠血浆中CD13的变化值与血压呈负相关(r=-0.758,P0.05);SHR大鼠血浆中AngⅢ的变化值与血压呈正相关(r=0.711,P0.05); SHR大鼠血浆中AngⅣ的变化值与血压呈负相关(r=-0.732,P0.05)；且SHR大鼠血浆中CD13表达水平动态变化与AngⅢ表达水平的动态变化亦具有负相关性(r=-0.746, P0.05); SHR大鼠血浆中CD13表达水平动态变化与AngⅣ表达水平的动态变化亦具有正相关性(r=0.698,P0.05)。HE和MASSON染色结果显示,SHR大鼠肾脏微血管周围、肾小管间质胶原沉积多于野生对照WKY大鼠,在10周、23周差异明显。 结论：CD13在肾脏中和循环系统中可能具有不同的生物学功能,参与调控机体的血压及其病理改变。
[Abstract]:Objective: to investigate the expression of CD13 in essential hypertension and its role in the development of hypertension.Methods Thirty two Wistar Kyoto (WKYY) rats (WKYY) and 32 Wistar Kyoto rats (WKYY) were randomly divided into 4 groups (8 rats in each group).The experimental observation was carried out at 4 weeks and 6 weeks (the onset of hypertension was 10 weeks) and the target organ damage stage was 23 weeks (target organ damage phase).The blood pressure of SHR-WKY was dynamically monitored by non-invasive caudal artery blood pressure meter.The 24 hours urine protein of the above mentioned rats was dynamically detected by ELISA method and the total RNAs of kidney tissue were extracted by Trizol method, and the expression of CD13mRNA in the kidney was dynamically detected by reverse transcriptase reverse transcription-polymerase chain reaction (RT-PCR), and the thickness of 2mm in the medulla longitudinalis of renal cortex of the rats in each group was obtained.Paraffin method was used to detect the expression of CD13 protein by immunohistochemical technique.The expression of CD13 Ang 鈪，

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