去肾交感神经对青年自发性高血压大鼠血压及靶器官的影响

发布时间：2018-01-01 09:04

  本文关键词：去肾交感神经对青年自发性高血压大鼠血压及靶器官的影响　出处：《河北医科大学》2016年硕士论文　论文类型：学位论文

  更多相关文章： 去肾交感神经 自发性高血压大鼠 血压 RAAS系统 左心室肥厚及纤维化 肾脏纤维化 主动脉硬化

【摘要】：目的:高血压作为一种全球流行性疾病,已有相关研究表明交感神经和肾素-血管紧张素-醛固酮系统(RAAS)的激活在高血压的发生和发展中起着极其重要的作用。应用药物可抑制交感神经和RAAS系统的过度激活以起到降压的作用,但需要长期服用,并存在药物副反应。近年来,国内外的研究表明,采用经皮导管去肾交感神经射频消融术(RFA)可有效的抑制交感神经和RAAS系统的过度激活使血压下降,达到治疗顽固性高血压的目的,进而使靶器官获益。由于这是一项治疗顽固性高血压的新技术,是在传统方法上的突破和创新,需要大量的基础以及临床研究来证明其安全有效性。已有研究显示该项技术可以使得自发性高血压大鼠(SHR)的血压下降将近40mm Hg。本研究旨在观察SHR去肾交感神经手术前后血压的变化以及对靶器官的影响,探讨去肾交感神经术对SHR血压的干预效果及相关靶器官的影响,进而为治疗原发性高血压提供新的治疗方法。方法:将12周龄的青年SHR(n=24)作为研究对象,联合应用外科手术切除及化学消融方法(10%的苯酚酒精溶液)建立去肾交感神经模型。将其随机分为去肾交感神经术组(RDN组)和假手术组(Sham组)。相同周龄的WKY大鼠(n=12)为基线对照组。术前及术后每周测量一次血压及体重至处死前一天。并分别于术后1w、8w、12w随机处死各组大鼠各4只,分别留取血浆、心脏、肾脏、主动脉。用ELISA法测定血浆中去甲肾上腺素(NE)及血管紧张素II(Ang II)的含量。计算左室质量指数(LVMI);心肌组织进行HE染色观察左心室心肌细胞形态;Western Blot法分别检测左心室α-MHC和COL I的蛋白表达。肾脏组织进行HE染色观察肾脏形态,进行PAS染色观察肾小球硬化程度;Western Blot法检测肾脏COL I的蛋白表达。主动脉进行HE染色观察主动脉形态,应用Image-Pro Plus 6.0测量血管内径(Luminal radius,LR)、血管中膜厚度(Media thickness,MT),计算血管中膜厚度/血管内径(MT/LR)。采用SPSS 13.0版软件进行统计分析,定量资料采用均数±标准差(?x±s)表示,两组均数间的比较采用t检验,多组均数间的比较采用单因素方差分析(one way ANOVA),多个时间点进行的重复测量采用重复测量方差分析。P0.05表示差异有统计学意义。结果:1体重的比较RDN组与Sham组在手术前及术后每周的体重比较无统计学差异(P0.05)。但RDN组与Sham组在手术前及术后每周的体重均低于WKY组(P0.05)。2各组收缩压的测量术前RDN组与Sham组的SBP无统计学差异(175.0±2.8mm Hg vs.174.5±2.7mm Hg)(P0.05),且均大于WKY组的SBP(132.2±4.2mm Hg)(P0.05)。与Sham组相比,RDN组的SBP在术后1w、2w、3w、4w、5w均低于Sham组(145.3±1.6mm Hg vs.181.0±3.7mm Hg,154.8±2.9mm Hg vs.190.4±7.7mm Hg,160.3±4.5mm Hg vs.196.0±6.4mm Hg,166.2±4.3mm Hg vs.203.6±8.0mm Hg,184.6±3.2mm Hg vs.201.5±9.0mm Hg)(P0.05),术后6w及以后各时间点两组SBP均无明显差异(P0.05)。与WKY组相比,RDN组和Sham组各对应时间点的SBP均高于WKY组(P0.05)。Sham组的SBP术后1w较术前稍升高(181.0±3.7mm Hg vs.174.5±2.7mm Hg)(P0.05),术后2w(190.4±7.7mm Hg)与术后1w无明显变化(P0.05),术后3w(196.0±6.4mm Hg)较术后2w稍升高(P0.05),术后3w以后SBP一直维持在较高水平且无明显变化(P0.05)。WKY组各个时间点的SBP均无明显变化(P0.05)。3血浆NE含量的变化术后1w RDN组血浆中NE含量与WKY组相比无统计学差异(169.1±3.8pg/ml vs.170.4±3.2pg/ml)(P0.05),且二者均显著低于Sham组(222.3±4.2pg/ml)(P0.05)。术后8w RDN组血浆中NE含量与Sham组相比无统计学差异(231.4±3.8pg/ml vs.234.5±2.7pg/ml)(P0.05),且均高于WKY组(171.3±5.6pg/ml)(P0.05)。术后12w RDN组血浆中NE含量与Sham组相比无统计学差异(238.6±2.7pg/ml vs.241.1±5.9pg/ml)(P0.05),且均高于WKY组(171.4±5.4pg/ml)(P0.05)。术后1w RDN组血浆中Ang II含量与WKY组相比无统计学差异(90.0±1.4pg/ml vs.89.3±2.4pg/ml)(P0.05),且二者均显著低于Sham组(171.9±2.8pg/ml)(P0.05)。术后8w RDN组血浆中Ang II含量与Sham组相比无统计学差异(174.5±2.0pg/ml vs.173.9±1.5pg/ml)(P0.05),且均高于WKY组(90.1±3.2pg/ml)(P0.05)。术后12w RDN组血浆中Ang II含量与Sham组相比无统计学差异(173.5±1.9pg/ml vs.173.3±2.8pg/ml)(P0.05),且均高于WKY组(90.1±2.9pg/ml)(P0.05)。5 LVMI的变化RDN组与Sham组相比术后1w、8w、12w的LVMI均无统计学差(P0.05),但上述两组LVMI在各自时间点均显著高于相应WKY组(P0.05)。6心肌HE染色观察三组大鼠心肌细胞形态学变化WKY组术后1w、8w、12w的心肌细胞大小均正常,排列整齐,心肌细胞核呈椭圆形,居于细胞中央,心肌间质无明显增生表现。RDN组与Sham组术后1w、8w、12w的心肌细胞均出现不同程度的损伤,表现为心肌细胞肥大且排列较紊乱,心肌间质明显水肿,部分心肌细胞溶解,周围结构不清。7 Western Blot法检测大鼠心脏α-MHC以及COL I的蛋白表达含量与Sham组相比,RDN组术后1w、8w、12wα-MHC的蛋白表达含量无统计学差异(P0.05),且均低于相同时间点WKY组的蛋白表达含量(P0.05)。与Sham组相比,RDN组术后1w、8w、12w COL I的蛋白表达含量无统计学差异(P0.05),且均高于相同时间点WKY组的蛋白表达含量(P0.05)。8肾脏HE染色及PAS染色观察三组大鼠肾脏形态学变化及肾小球硬化WKY组术后1w、8w、12w的肾小球大小及形态均正常,肾小管排列规则。RDN组与Sham组术后1w、8w、12w的个别肾小球皱缩甚至硬化,部分肾小球可见代偿性肥大,部分肾小管萎缩。9 Western Blot法检测肾脏COL I的蛋白表达含量与Sham组相比,RDN组术后1w、8w、12w COL I的蛋白表达含量无统计学差异(P0.05),且均高于相同时间点的WKY组的蛋白表达含4血浆中Ang II含量的变化量(P0.05)。10主动脉HE染色观察三组大鼠主动脉形态学变化及相关指标的测量WKY组的主动脉血管内径(LR)、中膜厚度(MT)及MT/LR在术后1w、8w、12w无明显变化(P0.05),且内径及中膜厚度均正常,血管壁细胞排列规则。Sham组的主动脉血管内径(LR)、中膜厚度(MT)及MT/LR在术后1w、8w、12w逐渐增大(P0.05),且血管壁细胞排列不规则,细胞密度减低。RDN组的主动脉血管内径(LR)、中膜厚度(MT)及MT/LR在术后1w、8w、12w逐渐增大(P0.05),且与相同时间点的Sham组无明显差别(P0.05),均大于相同时间点的WKY组(P0.05)。结论:1去肾交感神经术对大鼠的生长无影响,但长期高血压则阻碍大鼠的生长。2去肾交感神经术有降压的作用,尤其是术后1w血压明显下降,但其降压效果并不能稳定而持久。3去肾交感神经术降低血压的机制在于NE分泌的减少以及对RAAS系统的抑制。4由于去肾交感神经术仅能短暂降压,而不能持久、稳定的降压,因此高血压相关的靶器官的损伤也未能得到明显的改善。
[Abstract]:Objective: hypertension is a global epidemic, related studies have shown that the sympathetic and renin angiotensin aldosterone system (RAAS) activation plays an important role in the occurrence and development of hypertension. Application of drugs can inhibit the excessive activation of sympathetic nervous system in RAAS and to lower blood pressure however, the need for long-term use, and adverse drug reactions. In recent years, domestic and foreign research shows that the renal sympathetic nerve radiofrequency ablation percutaneous catheter (RFA) can effectively inhibit the sympathetic nervous system and RAAS excessive activation of decreased blood pressure, to achieve the purpose of treatment of refractory hypertension, so as to make the target organ benefit. Because this is a new technique for the treatment of refractory hypertension, is in the traditional method on the breakthrough and innovation, a large number of basic and clinical studies are needed to prove its safety and effectiveness. Studies have shown that the Technology can make the spontaneously hypertensive rats (SHR) of the blood pressure dropped nearly 40mm Hg. this study is to investigate the effects of SHR to the change of blood pressure before and after surgery and renal sympathetic nerve effects on target organs, explore the effect of intervention effect of renal denervation on the blood pressure of SHR and related target organs, and for the treatment of primary hypertension provide a new treatment method. Methods: 12 week old young SHR (n=24) as the research object, combined with the application of surgical resection and ablation method (alcohol solution of phenol 10%) to establish the renal sympathetic nerve model. The random divided into renal denervation group (RDN group) and sham operation group (group Sham). The same week old WKY rats (n=12) as the baseline control group. Preoperative and postoperative blood pressure and body weight was measured once a week to put to death the day before and after operation respectively. 1W, 8W, 12W were randomly sacrificed rats 4 rats each, were collected in plasma, Heart, kidney and aorta. Norepinephrine plasma was determined by ELISA method in (NE) and angiotensin II (Ang II) were calculated. The left ventricular mass index (LVMI); myocardial tissue morphological observation of left ventricular myocardial cells with HE staining; Western Blot method was used to detect left ventricular alpha -MHC and COL I the protein expression of renal tissues were observed by HE staining. Renal morphology was observed, the degree of glomerular sclerosis PAS staining; the expression of Western Blot renal COL I protein. HE staining was observed in aorta aortic morphology, using Image-Pro Plus 6 measuring blood tube diameter (Luminal radius, LR), blood vessels (Media thickness, MT film thickness), calculate the vascular thickness / vascular diameter (MT/LR). Statistical analysis was performed using SPSS software version 13, the quantitative data using the mean and standard deviation (? X + s) said that the two groups were compared using the mean t test, comparison of mean among groups. 閲囩敤鍗曞洜绱犳柟宸�鍒嗘灒�

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