本文關(guān)鍵詞：去腎交感神經(jīng)對(duì)青年自發(fā)性高血壓大鼠血壓及靶器官的影響　出處：《河北醫(yī)科大學(xué)》2016年碩士論文　論文類型：學(xué)位論文

  更多相關(guān)文章： 去腎交感神經(jīng) 自發(fā)性高血壓大鼠 血壓 RAAS系統(tǒng) 左心室肥厚及纖維化 腎臟纖維化 主動(dòng)脈硬化

【摘要】：目的:高血壓作為一種全球流行性疾病,已有相關(guān)研究表明交感神經(jīng)和腎素-血管緊張素-醛固酮系統(tǒng)(RAAS)的激活在高血壓的發(fā)生和發(fā)展中起著極其重要的作用。應(yīng)用藥物可抑制交感神經(jīng)和RAAS系統(tǒng)的過度激活以起到降壓的作用,但需要長期服用,并存在藥物副反應(yīng)。近年來,國內(nèi)外的研究表明,采用經(jīng)皮導(dǎo)管去腎交感神經(jīng)射頻消融術(shù)(RFA)可有效的抑制交感神經(jīng)和RAAS系統(tǒng)的過度激活使血壓下降,達(dá)到治療頑固性高血壓的目的,進(jìn)而使靶器官獲益。由于這是一項(xiàng)治療頑固性高血壓的新技術(shù),是在傳統(tǒng)方法上的突破和創(chuàng)新,需要大量的基礎(chǔ)以及臨床研究來證明其安全有效性。已有研究顯示該項(xiàng)技術(shù)可以使得自發(fā)性高血壓大鼠(SHR)的血壓下降將近40mm Hg。本研究旨在觀察SHR去腎交感神經(jīng)手術(shù)前后血壓的變化以及對(duì)靶器官的影響,探討去腎交感神經(jīng)術(shù)對(duì)SHR血壓的干預(yù)效果及相關(guān)靶器官的影響,進(jìn)而為治療原發(fā)性高血壓提供新的治療方法。方法:將12周齡的青年SHR(n=24)作為研究對(duì)象,聯(lián)合應(yīng)用外科手術(shù)切除及化學(xué)消融方法(10%的苯酚酒精溶液)建立去腎交感神經(jīng)模型。將其隨機(jī)分為去腎交感神經(jīng)術(shù)組(RDN組)和假手術(shù)組(Sham組)。相同周齡的WKY大鼠(n=12)為基線對(duì)照組。術(shù)前及術(shù)后每周測(cè)量一次血壓及體重至處死前一天。并分別于術(shù)后1w、8w、12w隨機(jī)處死各組大鼠各4只,分別留取血漿、心臟、腎臟、主動(dòng)脈。用ELISA法測(cè)定血漿中去甲腎上腺素(NE)及血管緊張素II(Ang II)的含量。計(jì)算左室質(zhì)量指數(shù)(LVMI);心肌組織進(jìn)行HE染色觀察左心室心肌細(xì)胞形態(tài);Western Blot法分別檢測(cè)左心室α-MHC和COL I的蛋白表達(dá)。腎臟組織進(jìn)行HE染色觀察腎臟形態(tài),進(jìn)行PAS染色觀察腎小球硬化程度;Western Blot法檢測(cè)腎臟COL I的蛋白表達(dá)。主動(dòng)脈進(jìn)行HE染色觀察主動(dòng)脈形態(tài),應(yīng)用Image-Pro Plus 6.0測(cè)量血管內(nèi)徑(Luminal radius,LR)、血管中膜厚度(Media thickness,MT),計(jì)算血管中膜厚度/血管內(nèi)徑(MT/LR)。采用SPSS 13.0版軟件進(jìn)行統(tǒng)計(jì)分析,定量資料采用均數(shù)±標(biāo)準(zhǔn)差(?x±s)表示,兩組均數(shù)間的比較采用t檢驗(yàn),多組均數(shù)間的比較采用單因素方差分析(one way ANOVA),多個(gè)時(shí)間點(diǎn)進(jìn)行的重復(fù)測(cè)量采用重復(fù)測(cè)量方差分析。P0.05表示差異有統(tǒng)計(jì)學(xué)意義。結(jié)果:1體重的比較RDN組與Sham組在手術(shù)前及術(shù)后每周的體重比較無統(tǒng)計(jì)學(xué)差異(P0.05)。但RDN組與Sham組在手術(shù)前及術(shù)后每周的體重均低于WKY組(P0.05)。2各組收縮壓的測(cè)量術(shù)前RDN組與Sham組的SBP無統(tǒng)計(jì)學(xué)差異(175.0±2.8mm Hg vs.174.5±2.7mm Hg)(P0.05),且均大于WKY組的SBP(132.2±4.2mm Hg)(P0.05)。與Sham組相比,RDN組的SBP在術(shù)后1w、2w、3w、4w、5w均低于Sham組(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),術(shù)后6w及以后各時(shí)間點(diǎn)兩組SBP均無明顯差異(P0.05)。與WKY組相比,RDN組和Sham組各對(duì)應(yīng)時(shí)間點(diǎn)的SBP均高于WKY組(P0.05)。Sham組的SBP術(shù)后1w較術(shù)前稍升高(181.0±3.7mm Hg vs.174.5±2.7mm Hg)(P0.05),術(shù)后2w(190.4±7.7mm Hg)與術(shù)后1w無明顯變化(P0.05),術(shù)后3w(196.0±6.4mm Hg)較術(shù)后2w稍升高(P0.05),術(shù)后3w以后SBP一直維持在較高水平且無明顯變化(P0.05)。WKY組各個(gè)時(shí)間點(diǎn)的SBP均無明顯變化(P0.05)。3血漿NE含量的變化術(shù)后1w RDN組血漿中NE含量與WKY組相比無統(tǒng)計(jì)學(xué)差異(169.1±3.8pg/ml vs.170.4±3.2pg/ml)(P0.05),且二者均顯著低于Sham組(222.3±4.2pg/ml)(P0.05)。術(shù)后8w RDN組血漿中NE含量與Sham組相比無統(tǒng)計(jì)學(xué)差異(231.4±3.8pg/ml vs.234.5±2.7pg/ml)(P0.05),且均高于WKY組(171.3±5.6pg/ml)(P0.05)。術(shù)后12w RDN組血漿中NE含量與Sham組相比無統(tǒng)計(jì)學(xué)差異(238.6±2.7pg/ml vs.241.1±5.9pg/ml)(P0.05),且均高于WKY組(171.4±5.4pg/ml)(P0.05)。術(shù)后1w RDN組血漿中Ang II含量與WKY組相比無統(tǒng)計(jì)學(xué)差異(90.0±1.4pg/ml vs.89.3±2.4pg/ml)(P0.05),且二者均顯著低于Sham組(171.9±2.8pg/ml)(P0.05)。術(shù)后8w RDN組血漿中Ang II含量與Sham組相比無統(tǒng)計(jì)學(xué)差異(174.5±2.0pg/ml vs.173.9±1.5pg/ml)(P0.05),且均高于WKY組(90.1±3.2pg/ml)(P0.05)。術(shù)后12w RDN組血漿中Ang II含量與Sham組相比無統(tǒng)計(jì)學(xué)差異(173.5±1.9pg/ml vs.173.3±2.8pg/ml)(P0.05),且均高于WKY組(90.1±2.9pg/ml)(P0.05)。5 LVMI的變化RDN組與Sham組相比術(shù)后1w、8w、12w的LVMI均無統(tǒng)計(jì)學(xué)差(P0.05),但上述兩組LVMI在各自時(shí)間點(diǎn)均顯著高于相應(yīng)WKY組(P0.05)。6心肌HE染色觀察三組大鼠心肌細(xì)胞形態(tài)學(xué)變化WKY組術(shù)后1w、8w、12w的心肌細(xì)胞大小均正常,排列整齊,心肌細(xì)胞核呈橢圓形,居于細(xì)胞中央,心肌間質(zhì)無明顯增生表現(xiàn)。RDN組與Sham組術(shù)后1w、8w、12w的心肌細(xì)胞均出現(xiàn)不同程度的損傷,表現(xiàn)為心肌細(xì)胞肥大且排列較紊亂,心肌間質(zhì)明顯水腫,部分心肌細(xì)胞溶解,周圍結(jié)構(gòu)不清。7 Western Blot法檢測(cè)大鼠心臟α-MHC以及COL I的蛋白表達(dá)含量與Sham組相比,RDN組術(shù)后1w、8w、12wα-MHC的蛋白表達(dá)含量無統(tǒng)計(jì)學(xué)差異(P0.05),且均低于相同時(shí)間點(diǎn)WKY組的蛋白表達(dá)含量(P0.05)。與Sham組相比,RDN組術(shù)后1w、8w、12w COL I的蛋白表達(dá)含量無統(tǒng)計(jì)學(xué)差異(P0.05),且均高于相同時(shí)間點(diǎn)WKY組的蛋白表達(dá)含量(P0.05)。8腎臟HE染色及PAS染色觀察三組大鼠腎臟形態(tài)學(xué)變化及腎小球硬化WKY組術(shù)后1w、8w、12w的腎小球大小及形態(tài)均正常,腎小管排列規(guī)則。RDN組與Sham組術(shù)后1w、8w、12w的個(gè)別腎小球皺縮甚至硬化,部分腎小球可見代償性肥大,部分腎小管萎縮。9 Western Blot法檢測(cè)腎臟COL I的蛋白表達(dá)含量與Sham組相比,RDN組術(shù)后1w、8w、12w COL I的蛋白表達(dá)含量無統(tǒng)計(jì)學(xué)差異(P0.05),且均高于相同時(shí)間點(diǎn)的WKY組的蛋白表達(dá)含4血漿中Ang II含量的變化量(P0.05)。10主動(dòng)脈HE染色觀察三組大鼠主動(dòng)脈形態(tài)學(xué)變化及相關(guān)指標(biāo)的測(cè)量WKY組的主動(dòng)脈血管內(nèi)徑(LR)、中膜厚度(MT)及MT/LR在術(shù)后1w、8w、12w無明顯變化(P0.05),且內(nèi)徑及中膜厚度均正常,血管壁細(xì)胞排列規(guī)則。Sham組的主動(dòng)脈血管內(nèi)徑(LR)、中膜厚度(MT)及MT/LR在術(shù)后1w、8w、12w逐漸增大(P0.05),且血管壁細(xì)胞排列不規(guī)則,細(xì)胞密度減低。RDN組的主動(dòng)脈血管內(nèi)徑(LR)、中膜厚度(MT)及MT/LR在術(shù)后1w、8w、12w逐漸增大(P0.05),且與相同時(shí)間點(diǎn)的Sham組無明顯差別(P0.05),均大于相同時(shí)間點(diǎn)的WKY組(P0.05)。結(jié)論:1去腎交感神經(jīng)術(shù)對(duì)大鼠的生長無影響,但長期高血壓則阻礙大鼠的生長。2去腎交感神經(jīng)術(shù)有降壓的作用,尤其是術(shù)后1w血壓明顯下降,但其降壓效果并不能穩(wěn)定而持久。3去腎交感神經(jīng)術(shù)降低血壓的機(jī)制在于NE分泌的減少以及對(duì)RAAS系統(tǒng)的抑制。4由于去腎交感神經(jīng)術(shù)僅能短暫降壓,而不能持久、穩(wěn)定的降壓,因此高血壓相關(guān)的靶器官的損傷也未能得到明顯的改善。
[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垎鏋，

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