本文選題：5HT1B　切入點(diǎn)：NF-κB通路　出處：《昆明醫(yī)科大學(xué)》2017年碩士論文

【摘要】：[目的]探討重組質(zhì)粒對血管平滑肌細(xì)胞增殖的影響和血管平滑肌細(xì)胞NF-κB通路活化的影響因素。[方法]制作重組的5-HT1B質(zhì)粒,構(gòu)建具有不同5-HT1B受控表達(dá)水平的血管平滑肌細(xì)胞模型。根據(jù)各自不同的干擾水平將細(xì)胞模型分成:完全空白對照組、高劑量脂質(zhì)體組(20ul)、低劑量脂質(zhì)體組(5ul)和5HT1B質(zhì)粒組,待培養(yǎng)條件穩(wěn)定后利用蛋白免疫印跡法測定各組VSMC的p42/44MAPK、PKC、IKK-α蛋白表達(dá)量,采用Tanon GIS圖像分析軟件分析各蛋白的表達(dá)水平。[結(jié)果]血管平滑肌細(xì)胞培養(yǎng)及轉(zhuǎn)染鑒定:用熒光顯微鏡(×400)下觀察5HT1B重組質(zhì)粒轉(zhuǎn)染VSMC48h后,發(fā)現(xiàn)綠色熒光位于胞漿中,說明重組質(zhì)粒已成功轉(zhuǎn)染進(jìn)入人VSMC;高劑量脂質(zhì)體處理的VSMC細(xì)胞內(nèi)IKK-α蛋白的表達(dá)被抑制(p0.001),低劑量處理的VSMC細(xì)胞與未經(jīng)任何處理的VSMC細(xì)胞(完全培養(yǎng)基組)的IKK-α蛋白水平無統(tǒng)計(jì)學(xué)差異(p0.05);未經(jīng)任何處理的VSMC細(xì)胞內(nèi)的PKC蛋白的表達(dá)量與高劑量處理的VSMC細(xì)胞內(nèi)PKC表達(dá)無統(tǒng)計(jì)學(xué)差異(p0.05),但低劑量脂質(zhì)體處理組的PKC表達(dá)顯著增加(p0.01); 5-HT1B質(zhì)粒組、高劑量脂質(zhì)體處理組以及低劑量脂質(zhì)體處理組的p42/44MAPK蛋白的表達(dá)量均明顯低于完全空白對照組(完全培養(yǎng)基組)(p0.001),且5-HT1B質(zhì)粒組的p42/44MAPK蛋白減少程度高于與高劑量脂質(zhì)體組和脂質(zhì)體組。[結(jié)論]發(fā)現(xiàn)血管平滑肌細(xì)胞MAPK通路蛋白轉(zhuǎn)染質(zhì)粒組有明顯低于各組的趨勢,5HTIB受體可能影響血管平滑肌細(xì)胞MAPK通路活化;未發(fā)現(xiàn)血管平滑肌細(xì)胞PKC及IKK-α蛋白的實(shí)驗(yàn)組有低于各組的趨勢,5HTIB受體可能不參與血管平滑肌細(xì)胞NF-κB通路活化;發(fā)現(xiàn)高劑量脂質(zhì)體組的IKKα和MAPK蛋白水平均被顯著抑制,推測脂質(zhì)體濃度可能影響NF-κB通路和MAPK通路的活化。
[Abstract]:[objective] to investigate the effects of recombinant plasmids on the proliferation of vascular smooth muscle cells and the factors affecting the activation of NF- 魏 B. [methods] Recombinant 5-HT1B plasmids were prepared. Vascular smooth muscle cell models with different levels of controlled expression of 5-HT1B were constructed. According to different interference levels, the cell models were divided into three groups: blank control group, high dose liposome group, low dose liposome group and 5HT1B plasmid group. After stable culture conditions, the expression of p42 / 44 MAPK- 偽 protein in VSMC was determined by Western blot. Tanon GIS image analysis software was used to analyze the expression level of each protein. [results] VSMCs were cultured and identified. After transfection of VSMC48h with 5HT1B recombinant plasmid under fluorescence microscope (脳 400), it was found that the green fluorescence was located in the cytoplasm. These results indicate that the recombinant plasmid has been successfully transfected into human VSMC, the expression of IKK- 偽 protein was inhibited in VSMC cells treated with high dose liposome, and IKK- 偽 protein in VSMC cells treated with low dose and in VSMC cells without any treatment (complete medium group) was inhibited. The expression of PKC protein in VSMC cells without any treatment was not significantly different from that in VSMC cells treated with high dose, but the expression of PKC in low dose liposome treatment group was significantly higher than that in 5-HT1B plasmid group. The expression of p42/44MAPK protein in the high dose liposome treatment group and the low dose liposome treatment group was significantly lower than that in the complete blank control group (complete culture medium group), and the decrease of p42/44MAPK protein in the 5-HT1B plasmid group was higher than that in the high dose liposome group. [conclusion] it was found that the MAPK pathway activation of vascular smooth muscle cells in plasmid group was significantly lower than that in plasmid group (P < 0.05). PKC and IKK- 偽 protein in vascular smooth muscle cells in experimental group were lower than those in each group. 5 HTIB receptor may not participate in the activation of NF- 魏 B pathway in vascular smooth muscle cells, and the levels of IKK 偽 and MAPK protein in high dose liposome group were significantly inhibited. It is speculated that the concentration of liposome may affect the activation of NF- 魏 B pathway and MAPK pathway.
【學(xué)位授予單位】：昆明醫(yī)科大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2017
【分類號】：R135

【參考文獻(xiàn)】

相關(guān)期刊論文 前10條

1 鄭創(chuàng)亮;許丹;曹獻(xiàn)磅;馮秀瓊;;某高爾夫球桿生產(chǎn)企業(yè)93例職業(yè)性手臂振動病調(diào)查報(bào)告[J];職業(yè)衛(wèi)生與應(yīng)急救援;2015年04期

2 魏鈞伯;馬健;陸志剛;馬士新;趙鋼;蔡忷繩;魏盟;;血管緊張素Ⅱ?qū)ρ芷交〖?xì)胞血管緊張素Ⅱ受體1、2mRNA表達(dá)的影響[J];臨床心血管病雜志;2015年02期

3 黃永順;金佳純;溫賢忠;吳勁松;伍津;胡世杰;;2006—2012年廣東省物理因素所致職業(yè)病發(fā)病特點(diǎn)分析和對策探討[J];中國職業(yè)醫(yī)學(xué);2013年06期

4 陳青松;李文;郎麗;肖斌;李宏玲;徐國勇;;5-羥色胺受體1B基因多態(tài)性及其與振動性白指關(guān)聯(lián)分析[J];中國職業(yè)醫(yī)學(xué);2013年01期

5 熊瑋;董少紅;袁建輝;李江華;劉建軍;徐新云;;MiRNA-146a通過NF-κB依賴的途徑促進(jìn)血管平滑肌細(xì)胞增殖[J];南方醫(yī)科大學(xué)學(xué)報(bào);2012年02期

6 鄭倩玲;楊愛初;陳嘉斌;夏麗華;曾子芳;李斌;;84例職業(yè)性手臂振動病病人手部末梢循環(huán)障礙臨床分析[J];中國職業(yè)醫(yī)學(xué);2010年04期

7 段超;陳鑫;邱志兵;許歡;;大鼠胸主動脈血管平滑肌細(xì)胞的原代培養(yǎng)和鑒定[J];臨床肺科雜志;2010年04期

8 鐘華;何芳;胡清華;王振煥;鄧峰美;孫志萍;李增春;;血管平滑肌細(xì)胞中ERK通路與TGF-β_1/Smad通路的相互作用[J];中國應(yīng)用生理學(xué)雜志;2010年01期

9 繆榮明;;中西醫(yī)聯(lián)合治療手臂振動病31例臨床觀察[J];中國實(shí)用醫(yī)藥;2010年05期

10 李冰冰;姜楨;;5-羥色胺在肺動脈平滑肌細(xì)胞增殖/肺血管重構(gòu)中的作用[J];國際麻醉學(xué)與復(fù)蘇雜志;2007年05期

，

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