發(fā)布時間：2018-03-23 09:02

  本文選題：代謝手術(shù)　切入點：糖尿病心肌病　出處：《山東大學(xué)》2017年碩士論文　論文類型：學(xué)位論文

【摘要】：[研究背景]糖脂代謝障礙引起的心肌功能紊亂是糖尿病的嚴(yán)重并發(fā)癥之一,目前尚無有效的治療方法。近年來,代謝手術(shù)越來越普遍用于醫(yī)治糖尿病及其并發(fā)癥。其在改善、緩解糖尿病心肌病(diabetic cardiomyopathy,DCM))方面的有效性亦日益受到人們的重視,但是其機(jī)制還不清楚。普遍認(rèn)為,內(nèi)質(zhì)網(wǎng)應(yīng)激(endoplasmic retjculum stress,ERS)介導(dǎo)的細(xì)胞凋亡與DCM的病理進(jìn)程息息相關(guān)。在糖尿病狀態(tài)下,心肌細(xì)胞糖脂代謝異常,內(nèi)質(zhì)網(wǎng)的工作環(huán)境受到干擾,ERS啟動欲糾正細(xì)胞的錯誤蛋白折疊,以維持細(xì)胞的正常功能。若干擾力量較大,ERS無法復(fù)原細(xì)胞的正常生理功能,便引發(fā)細(xì)胞凋亡程序,來維持機(jī)體的正常運(yùn)轉(zhuǎn)。為了研究代謝手術(shù)對DCM的緩解作用,探討其相關(guān)機(jī)制,本研究在DCM動物模型基礎(chǔ)上,明確代謝手術(shù)對糖尿病大鼠心肌功能的緩解作用,并通過對心肌細(xì)胞內(nèi)質(zhì)網(wǎng)應(yīng)激通路相關(guān)蛋白表達(dá)變化的比較與分析,進(jìn)一步探討代謝手術(shù)后DCM緩解的機(jī)制。[研究方法]本實驗以高脂飲食及小劑量鏈脲佐菌素(STZ)誘導(dǎo)的糖尿病大鼠為研究動物模型,分三組,分別施以十二指腸空腸旁路術(shù)(DJB)、袖狀胃切除術(shù)(SG)、及假手術(shù)(SHAM)。各組觀測指標(biāo)包括:(1)測量術(shù)前術(shù)后各特定時間點的體重及全身的血糖、脂質(zhì)水平;(2)行口服葡萄糖耐量實驗(Oral glucose tolerance test,0GTT)和胰島素耐量實驗(Insulin tolerance test,ITT),比較代謝手術(shù)對糖尿病大鼠糖代謝的影響;(3)術(shù)后8周,行超聲心動圖檢測,比較代謝手術(shù)對糖尿病大鼠心功能的影響情況;(4)行HE染色、Masson染色、油紅0染色和Tunel染色,測量比較代謝手術(shù)后,心肌的形態(tài)學(xué)變化及細(xì)胞凋亡情況;(5)行免疫組化實驗、wesetern blot 及qRT-PCR檢測ERS信號分子GRP78、PERK、p-PERK、iRE1α、p-IRE1a、ATF6、CHOP 和 caspase12 的表達(dá)情況。[研究結(jié)果]1、成功給予三組大鼠相應(yīng)手術(shù),在整個實驗過程中無大鼠死亡。2、術(shù)前,三組大鼠體重?zé)o差別;術(shù)后1周,三組大鼠體重下降至最低點,后逐步回升,DJB組與SHAM組大鼠體重回升幅度相當(dāng),在整個實驗周期中,兩組大鼠體重?zé)o統(tǒng)計學(xué)差別;SG組大鼠體重上升相對較緩,術(shù)后6周、8周,SG組大鼠體重與DJB組和SHAM組大鼠相比,差別達(dá)到統(tǒng)計學(xué)意義(P0.05)。3、術(shù)前,三組大鼠AUCOGTT與AUCITT無差別;術(shù)后2周、8周,DJB組與SG組大鼠AUCOGTT與AUCITT值顯著低于SHAM組(P0.01),DJB組與SG組間無差別。4、術(shù)后8周,DJB組與SG組大鼠心功能較SHAM組顯著改善,超聲心動圖結(jié)果表現(xiàn)為,DJB組與SG組LVEDd與LVPWd較SHAM組明顯降低(LVEDd:P0.01;LVPWd:P0.001),LVEF、FS、E/A 較 SHAM 組明顯升高 CLVEF:P0.01;FS:P0.05;E/A:P0.001);DJB 組與 SG 組間無差別。5、HE及Masson染色結(jié)果提示,代謝手術(shù)后,心肌細(xì)胞肥大、心肌纖維化情況較SHAM組緩解;油紅0染色結(jié)果提示代謝手術(shù)不影響心肌細(xì)胞的脂質(zhì)沉積。6、術(shù)后8周,DJB組與SG組大鼠心肌細(xì)胞凋亡率顯著低于SHAM組(P0.001),DJB組與SG組間無差別。7、免疫組化、western blot及qRT-PCR結(jié)果均提示,代謝手術(shù)后,GRP78、CHOP、caspasel2及PERK、p-PERK明顯低于SHAM組,DJB組與SG組間差別無統(tǒng)計學(xué)意義;三組間IRE1、p-IRE1及ATF6表達(dá)均無統(tǒng)計學(xué)差別。[研究結(jié)論]1、DJB與SG均可有效的改善糖尿病大鼠的糖耐量及胰島素抵抗,緩解糖尿病大鼠的心肌功能。2、DJB與SG通過抑制PERK途徑抑制糖尿病大鼠心肌細(xì)胞內(nèi)質(zhì)網(wǎng)應(yīng)激,降低心肌細(xì)胞凋亡,從而改善心肌功能。3、DJB與SG在改善糖尿病大鼠心肌功能紊亂方面無差別。
[Abstract]:[background] myocardial dysfunction caused by lipid metabolism disorder is a serious complication of diabetes, there is no effective treatment. In recent years, more and more widely used in the treatment of metabolic surgery of diabetes and its complications. The improvement in ease of diabetic cardiomyopathy (diabetic) cardiomyopathy, DCM) effectiveness also has been paid more and more attention however, the mechanism is not clear. It is generally accepted that the endoplasmic reticulum stress (endoplasmic retjculum, stress, ERS) pathological process and cell apoptosis mediated by DCM are closely related. In the diabetic state, myocardial cell lipid metabolism, endoplasmic reticulum work environment is disturbed, ERS starting cell error correct protein folding, to to maintain the normal function of the cells. If the interference power is larger, the normal physiological function of ERS cells cannot be restored, will lead to cell apoptosis process, to maintain the body's normal In order to alleviate the effects of the operation. The metabolic surgery for DCM, to explore the underlying mechanism, based on the research of DCM animal model, clear metabolic surgery on myocardial function of diabetic rats relieving, comparison and analysis and expression of myocardial cells through endoplasmic reticulum stress related protein, to further explore the mechanism of metabolic research methods. After the operation of DCM mitigation] in this experiment, high fat diet and a low dose of streptozotocin (STZ) induced diabetic rats as animal model study, divided into three groups, were treated with duodenal jejunal bypass (DJB), sleeve gastrectomy (SG), and sham operation (SHAM). Each group included observation index: (1) blood glucose, body weight and the specific point in time measurement of preoperative and postoperative lipid level; (2) underwent oral glucose tolerance test (Oral glucose tolerance test, 0GTT) and insulin tolerance test (Insulin tolerance test, ITT ), to compare the effects of metabolic surgery on glucose metabolism in diabetic rats; (3) 8 weeks after operation, echocardiography detection, comparison of metabolic surgery on cardiac function in diabetic rats; (4) by HE staining, Masson staining, oil red 0 and Tunel staining, measurement of metabolic surgery after the morphologic changes and apoptosis in myocardium; (5) immunohistochemical experiments, ERS Wesetern and qRT-PCR blot detection signal molecules GRP78, PERK, p-PERK, p-IRE1a, iRE1 alpha, ATF6, the expression of CHOP and caspase12. On]1, the success of the three groups of rats given the corresponding operation in the whole experiment in the process of no rat died in.2, before the operation, the weight of rats in the three groups no difference; after 1 weeks, body weight of rats in the three groups decreased to the lowest point, after a gradual recovery in DJB group and SHAM group rats weight recovery rate is in the whole experimental period, no significant difference of body weight of rats in the two groups group SG rats; 閲嶄笂鍗囩浉瀵硅緝緙，

本文編號：1652788