本文選題：CD137　切入點(diǎn)：自噬　出處：《江蘇大學(xué)》2017年碩士論文

【摘要】：目的 探討CD137-CD137L信號(hào)是否通過(guò)激活細(xì)胞自噬影響內(nèi)皮細(xì)胞管腔形成及動(dòng)脈環(huán)血管新生。方法1.構(gòu)建C57BL/6J小鼠胸主動(dòng)脈動(dòng)脈環(huán)模型,在倒置相差顯微鏡下觀察:(1)CD137-CD137L信號(hào)對(duì)動(dòng)脈環(huán)出芽長(zhǎng)度的影響;(2)干預(yù)自噬對(duì)CD137-CD137L信號(hào)介導(dǎo)的動(dòng)脈環(huán)出芽長(zhǎng)度的影響;2.基質(zhì)膠上培養(yǎng)人臍靜脈內(nèi)皮細(xì)胞(human umbilical vein endothelial cells,HUVECs),倒置相差顯微鏡下動(dòng)態(tài)觀察:(1)CD137-CD137L信號(hào)對(duì)內(nèi)皮細(xì)胞管腔形成能力的影響;(2)干預(yù)自噬對(duì)CD137-CD137L信號(hào)介導(dǎo)的內(nèi)皮細(xì)胞管腔形成能力的影響;3.運(yùn)用蛋白質(zhì)免疫印記(Western blot)技術(shù)檢測(cè)干預(yù)CD137-CD137L信號(hào)對(duì)自噬關(guān)鍵蛋白分子LC3 II/I、P62表達(dá)的影響;4.利用自噬雙熒光(mRFP-GFP-LC3)標(biāo)記的腺病毒感染內(nèi)皮細(xì)胞,在熒光顯微鏡下觀察干預(yù)CD137-CD137L信號(hào)對(duì)胞漿自噬小體(黃色熒光)生成情況以及自噬小體與溶酶體融合(紅色熒光)情況的影響;5.運(yùn)用Transwell細(xì)胞遷移試驗(yàn),檢測(cè)干預(yù)自噬后CD137-CD137L信號(hào)介導(dǎo)的內(nèi)皮細(xì)胞遷移能力的變化;6.采用CCK-8細(xì)胞增殖試劑盒,檢測(cè)干預(yù)自噬后CD137-CD137L信號(hào)介導(dǎo)的內(nèi)皮細(xì)胞增殖能力的變化。結(jié)果1.激活CD137-CD137L信號(hào),HUVECs管腔形成總長(zhǎng)度增加(P0.05 vs對(duì)照組)、動(dòng)脈環(huán)出芽總長(zhǎng)度平均值較對(duì)照組顯著增加(P0.01 vs對(duì)照組);抑制CD137-CD137L信號(hào)HUVECs管腔形成總長(zhǎng)度減少(P0.05 vs刺激組)、動(dòng)脈環(huán)出芽總長(zhǎng)度平均值顯著減少(P0.01 vs刺激組);2.激動(dòng)CD137-CD137L信號(hào)內(nèi)皮細(xì)胞自噬關(guān)鍵分子LC3II蛋白表達(dá)增加,且P62蛋白表達(dá)減少較對(duì)照組(P均0.05 vs對(duì)照組),同時(shí)自噬小體和自噬小體溶酶體均顯著增加(P均0.01 vs對(duì)照組);而抑制CD137-CD137L信號(hào),HUVECs LC3II蛋白表達(dá)下調(diào)、P62蛋白相對(duì)表達(dá)上調(diào)(P均0.05 vs刺激組),同時(shí)自噬小體和自噬小體溶酶體均顯著減少(P均0.01 vs刺激組);3.3-MA(5mM)抑制內(nèi)皮細(xì)胞自噬后,激動(dòng)CD137-CD137L信號(hào),HUVECs管腔形成總長(zhǎng)度減少(P0.05)且動(dòng)脈環(huán)出芽長(zhǎng)度平均值顯著減少(P0.01);抑制自噬后激活CD137-CD137L信號(hào),內(nèi)皮細(xì)胞遷移及增殖能力均顯著增加(P均0.01)。結(jié)論CD137-CD137L信號(hào)可能通過(guò)激活細(xì)胞自噬介導(dǎo)內(nèi)皮細(xì)胞管腔形成及動(dòng)脈環(huán)血管新生。
[Abstract]:Objective to investigate whether CD137-CD137L signal affects endothelial cell lumen formation and angiogenesis by activating autophagy. Methods 1. A model of thoracic aortic rings in C57BL/6J mice was established. Observation on the effect of CD137-CD137L signal on the sprouting length of arterial ring under inverted phase contrast microscope) intervention of autophagy on the length of CD137-CD137L signaling mediated arterial ring sprouting. 2. Cultured human umbilical vein endothelial cells on matrix glue, inverted phase. Dynamic observation of the effect of CD137-CD137L signal on Endothelial Cell Endothelial Cell Endothelial Cell cavities under differential microscope. The effect of autophagy on the Endothelial Cell Endothelial Cell Endothelial Cell Endothelial Cell Endothelial formation mediated by CD137-CD137L signal. The effect of protein Immunoimprint Western blot technique on the Endothelial Cell Endothelial cavity formation ability. The effect of signal on the expression of LC3 II / Igni P62, a key protein molecule in autophagy. (4) Adenovirus-labeled adenovirus was used to infect endothelial cells, which was labeled with mRFP-GFP-LC3. The effects of interference with CD137-CD137L signal on the formation of cytoplasmic autophagy (yellow fluorescence) and the fusion of autophagy and lysosome (red fluorescence) were observed under fluorescence microscope. The Transwell cell migration test was used. To detect the changes of endothelial cell migration mediated by CD137-CD137L signal after intervention of autophagy. The CCK-8 cell proliferation kit was used. The changes of endothelial cell proliferation mediated by CD137-CD137L signal after intervention were detected. Results 1.Activation of CD137-CD137L signal increased the total length of lumen formation of HUVECs (P0.05 vs control group), and the mean of total length of arterial ring bud increased significantly (P0.01 vs control group). 2. In control group, the total length of HUVECs lumen formation was decreased in CD137-CD137L signal inhibition group (P0.05) vs stimulation group (P 0.05), and the mean of total sprouting length of arterial ring decreased significantly (P 0.01 vs stimulation group) 2. The expression of LC3II protein, a key molecule of autophagy, was increased in activated CD137-CD137L signal endothelial cells. The expression of P62 protein was lower than that of control group (P 0.05 vs control group, P 0.05 vs control group), and that of autophagy and autophagy lysosomes were significantly increased (P 0.01 vs control group, respectively), while the down-regulation of P62 protein expression was down-regulated by inhibiting CD137-CD137L signal expression in HUVECs and upregulating the relative expression of P62 protein. P 0.05 vs stimulation group, while autophagy and autophagy lysosomes decreased significantly (P 0.01 vs stimulation group 3.3-MA-5mm) inhibited endothelial cell autophagy. The total length of lumen formation of HUVECs was reduced by P0.05) and the mean sprouting length of arterial ring was significantly reduced by P0.01C, and the activation of CD137-CD137L signal was inhibited after autophagy. Both migration and proliferation of endothelial cells increased significantly (P < 0.01). Conclusion CD137-CD137L signal may mediate endothelial cell lumen formation and arterial ring angiogenesis by activation of autophagy.
【學(xué)位授予單位】：江蘇大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2017
【分類號(hào)】：R54

【相似文獻(xiàn)】

相關(guān)期刊論文 前10條

1 郭守利;李倩;張一飛;劉曄;朱大嶺;;花生四烯酸及其3種代謝產(chǎn)物對(duì)兔肺動(dòng)脈環(huán)作用的比較[J];中國(guó)藥理學(xué)通報(bào);2006年06期

2 葉正中;;220例獼猴(Macaca mulatta)Willis氏動(dòng)脈環(huán)的解剖范型和異常的觀察[J];解剖學(xué)報(bào);1980年02期

3 李世校;張靜;張國(guó)榮;吳愛(ài)群;李延賓;;幾種實(shí)驗(yàn)動(dòng)物腦動(dòng)脈環(huán)的觀察[J];解剖學(xué)雜志;1993年03期

4 董雅占;;中國(guó)人腦動(dòng)脈環(huán)的初步觀察[J];解剖學(xué)報(bào);1958年01期

5 張成英;兔腦動(dòng)脈環(huán)的解剖學(xué)觀察[J];蚌埠醫(yī)學(xué)院學(xué)報(bào);1993年01期

6 張煥萍,徐永健,張珍祥,倪望,陳仕新;核因子-κB抑制劑在蛋白激酶C對(duì)低氧性肺動(dòng)脈高壓大鼠肺動(dòng)脈環(huán)反應(yīng)性影響中的作用[J];中國(guó)病理生理雜志;2004年02期

7 萬(wàn)有，，王迪潯;過(guò)氧化氫和次黃嘌呤＋黃嘌呤氧化酶對(duì)家兔離體肺、腦動(dòng)脈環(huán)缺氧反應(yīng)的影響[J];中國(guó)應(yīng)用生理學(xué)雜志;1994年04期

8 劉琰,徐永健,張珍祥;蛋白激酶C對(duì)大鼠離體肺動(dòng)脈環(huán)張力及其反應(yīng)性的調(diào)節(jié)作用[J];中國(guó)應(yīng)用生理學(xué)雜志;1999年03期

9 李玢琳;張錦;李娜;佟冬怡;;不同濃度依托咪酯對(duì)內(nèi)毒素孵育前后兔離體肺、體動(dòng)脈環(huán)張力的影響[J];實(shí)用藥物與臨床;2012年02期

10 ;動(dòng)脈環(huán)與單側(cè)聽覺(jué)癥狀之間有關(guān)聯(lián)嗎?[J];國(guó)外醫(yī)學(xué)(耳鼻咽喉科學(xué)分冊(cè));1999年06期

相關(guān)會(huì)議論文 前3條

1 張海波;董衛(wèi);徐志偉;蘇肇伉;丁文祥;;肺動(dòng)脈環(huán)束術(shù)治療復(fù)雜先天性心臟病[A];中華醫(yī)學(xué)會(huì)第七次全國(guó)胸心血管外科學(xué)術(shù)會(huì)議暨2007中華醫(yī)學(xué)會(huì)胸心血管外科青年醫(yī)師論壇論文集心血管外科分冊(cè)[C];2007年

2 佟冬怡;張錦;;去甲腎上腺素、多巴胺對(duì)內(nèi)毒素孵育離體肺、體動(dòng)脈環(huán)張力影響的比較[A];2009年全國(guó)危重病急救醫(yī)學(xué)學(xué)術(shù)會(huì)議論文匯編[C];2009年

3 張玉青;黃萍;陳醒言;龔永生;毛孫忠;胡良岡;洪林;范小芳;;Apelin對(duì)正常與低氧大鼠離體肺動(dòng)脈環(huán)的舒張作用及其機(jī)制[A];浙江省生理科學(xué)會(huì)2008年學(xué)術(shù)年會(huì)論文匯編[C];2008年

相關(guān)碩士學(xué)位論文 前2條

1 李波;CD137-CD137L信號(hào)通過(guò)激活自噬影響內(nèi)皮細(xì)胞管腔形成以及動(dòng)脈環(huán)血管新生[D];江蘇大學(xué);2017年

2 李靜;活性氧介導(dǎo)自發(fā)性高血壓大鼠離體血管的血管緊張素Ⅱ的縮血管效應(yīng)的研究[D];南京醫(yī)科大學(xué);2009年

本文編號(hào)：1664391