慢病毒轉(zhuǎn)染的β3-AR基因?qū)π募》屎竦挠绊?/H1>
發(fā)布時(shí)間：2018-06-28 01:21
本文選題：β腎上腺素能受體 + 心肌細(xì)胞 ； 參考：《臨床心血管病雜志》2017年01期
【摘要】：目的:探討β3腎上腺素能受體(β3-AR)對(duì)SD大鼠乳鼠心肌肥大的影響及其機(jī)制。方法:體外培養(yǎng)SD大鼠乳鼠心肌細(xì)胞,用攜帶β3-AR基因的慢病毒轉(zhuǎn)染細(xì)胞后,用去甲腎上腺素(NE)誘導(dǎo)細(xì)胞48h,建立心肌細(xì)胞肥大模型。實(shí)驗(yàn)分4組:空白對(duì)照組(Control組)、心肌肥厚組(NE組)、β3-AR基因轉(zhuǎn)染+NE組(β3-AR組)、空病毒轉(zhuǎn)染+NE組(空病毒組)。用免疫熒光法鑒定心肌細(xì)胞,倒置熒光顯微鏡觀察病毒轉(zhuǎn)染組綠色熒光蛋白(GFP)表達(dá),免疫印跡法(Western Blot)檢測(cè)β3-AR、絲裂原活化蛋白激酶p38(p38MAPK)、細(xì)胞外信號(hào)調(diào)控激酶(ERK1/2)、磷酸化p38MAPK(p-p38MAPK)和ERK(p-ERK1/2)及原癌基因c-myc、c-fos蛋白水平的表達(dá)。結(jié)果:(1)慢病毒介導(dǎo)β3-AR基因轉(zhuǎn)染心肌細(xì)胞,β3-AR表達(dá)較空白對(duì)照組明顯升高。(2)用Western Blot檢測(cè)各實(shí)驗(yàn)組細(xì)胞原癌基因c-myc、c-fos表達(dá),其中NE組、β3-AR組、空病毒組表達(dá)均高于空白對(duì)照組,其中β3-AR組cmyc、c-fos表達(dá)明顯高于NE組。(3)NE組、β3-AR組、空病毒組p38MAPK及ERK1/2的磷酸化水平較空白對(duì)照組明顯上調(diào),其中β3-AR組表達(dá)最高。結(jié)論:慢病毒介導(dǎo)的β3-AR基因轉(zhuǎn)染使心肌細(xì)胞有效高表達(dá)β3-AR,β3-AR可能通過(guò)MAPK通路促進(jìn)心肌肥厚。
[Abstract]:Aim: to investigate the effect of 尾 3 adrenoceptor (尾 3 AR) on myocardial hypertrophy in neonatal SD rats and its mechanism. Methods: neonatal SD rat cardiomyocytes were cultured in vitro. The cells were transfected with lentivirus carrying 尾 3-AR gene and induced by norepinephrine (NE) for 48h to establish cardiomyocyte hypertrophy model. The experiment was divided into four groups: control group (blank control group), NE group (myocardial hypertrophy group), NE group (尾 3-AR group) transfected with 尾 3-AR gene, and NE group (empty virus group) transfected with empty virus. The expression of green fluorescent protein (GFP) in viral transfection group was observed by reverse fluorescence microscope. Western blot was used to detect the expression of 尾 3-AR, p38 MAPK, ERK1 / 2, phosphorylated p38 MAPK and ERK (p-ERK1 / 2) and proto-oncogene c-mycc-fos. Results: (1) the expression of 尾 _ 3-AR gene was significantly higher than that of the blank control group. (2) the expression of proto-oncogene c-mycnc-fos was detected by Western blot, and the expression of c-mycfs in NE group, 尾 _ 3-AR group and empty virus group was higher than that in blank control group. The expression of c-fos in 尾 3-AR group was significantly higher than that in NE group. (3) the phosphorylation levels of p38 MAPK and ERK1 / 2 in NE group, 尾 3-AR group and empty virus group were significantly higher than those in control group, and the highest expression was found in 尾 3-AR group. Conclusion: lentivirus-mediated 尾 _ 3-AR gene transfection can effectively overexpression 尾 _ 3-AR. 尾 _ 3-AR may promote myocardial hypertrophy through MAPK pathway.
【作者單位】： 新疆石河子大學(xué)醫(yī)學(xué)院第一附屬醫(yī)院心內(nèi)二科;新疆醫(yī)科大學(xué)第一附屬醫(yī)院心臟中心;
【基金】：國(guó)家自然科學(xué)基金-地區(qū)科學(xué)基金項(xiàng)目(No:81260028) 石河子大學(xué)科學(xué)技術(shù)研究發(fā)展計(jì)劃“自然科學(xué)與技術(shù)創(chuàng)新”團(tuán)隊(duì)創(chuàng)新項(xiàng)目(No:2011ZRKXTD-07)
【分類號(hào)】：R541.6
【相似文獻(xiàn)】
相關(guān)期刊論文 前6條
1 樊守剛;吳小濤;王運(yùn)濤;李果;;骨髓間充質(zhì)干細(xì)胞經(jīng)慢病毒轉(zhuǎn)染綠色熒光蛋白后的體內(nèi)觀察[J];東南大學(xué)學(xué)報(bào)(醫(yī)學(xué)版);2010年02期
2 劉君;向川;衛(wèi)小春;;不同轉(zhuǎn)染方法對(duì)逆轉(zhuǎn)錄病毒轉(zhuǎn)染效率影響的研究[J];實(shí)用骨科雜志;2008年12期
3 王新武;劉文革;;人bFGF重組慢病毒轉(zhuǎn)染BMSCs的實(shí)驗(yàn)研究[J];中國(guó)現(xiàn)代醫(yī)生;2010年06期
4 方敏，，王立人，雷學(xué)鋒，畢愛(ài)華;逆轉(zhuǎn)錄病毒轉(zhuǎn)染的小鼠IL－3基因在NIH／3T3細(xì)胞中的表達(dá)[J];同濟(jì)醫(yī)科大學(xué)學(xué)報(bào);1996年06期
5 劉濤,李金蓮;大鼠三叉神經(jīng)脊束核尾側(cè)亞核淺層內(nèi)GABA樣、L-ENK樣、GlyT2樣陽(yáng)性末梢與Sindibis病毒轉(zhuǎn)染表達(dá)GFP神經(jīng)元之間的聯(lián)系[J];中國(guó)神經(jīng)科學(xué)雜志;2002年04期
6 ;[J];;年期
本文編號(hào)：2076083
本文鏈接：http://sikaile.net/kejilunwen/jiyingongcheng/2076083.html
本文選題：β腎上腺素能受體 + 心肌細(xì)胞 ； 參考：《臨床心血管病雜志》2017年01期
【摘要】：目的:探討β3腎上腺素能受體(β3-AR)對(duì)SD大鼠乳鼠心肌肥大的影響及其機(jī)制。方法:體外培養(yǎng)SD大鼠乳鼠心肌細(xì)胞,用攜帶β3-AR基因的慢病毒轉(zhuǎn)染細(xì)胞后,用去甲腎上腺素(NE)誘導(dǎo)細(xì)胞48h,建立心肌細(xì)胞肥大模型。實(shí)驗(yàn)分4組:空白對(duì)照組(Control組)、心肌肥厚組(NE組)、β3-AR基因轉(zhuǎn)染+NE組(β3-AR組)、空病毒轉(zhuǎn)染+NE組(空病毒組)。用免疫熒光法鑒定心肌細(xì)胞,倒置熒光顯微鏡觀察病毒轉(zhuǎn)染組綠色熒光蛋白(GFP)表達(dá),免疫印跡法(Western Blot)檢測(cè)β3-AR、絲裂原活化蛋白激酶p38(p38MAPK)、細(xì)胞外信號(hào)調(diào)控激酶(ERK1/2)、磷酸化p38MAPK(p-p38MAPK)和ERK(p-ERK1/2)及原癌基因c-myc、c-fos蛋白水平的表達(dá)。結(jié)果:(1)慢病毒介導(dǎo)β3-AR基因轉(zhuǎn)染心肌細(xì)胞,β3-AR表達(dá)較空白對(duì)照組明顯升高。(2)用Western Blot檢測(cè)各實(shí)驗(yàn)組細(xì)胞原癌基因c-myc、c-fos表達(dá),其中NE組、β3-AR組、空病毒組表達(dá)均高于空白對(duì)照組,其中β3-AR組cmyc、c-fos表達(dá)明顯高于NE組。(3)NE組、β3-AR組、空病毒組p38MAPK及ERK1/2的磷酸化水平較空白對(duì)照組明顯上調(diào),其中β3-AR組表達(dá)最高。結(jié)論:慢病毒介導(dǎo)的β3-AR基因轉(zhuǎn)染使心肌細(xì)胞有效高表達(dá)β3-AR,β3-AR可能通過(guò)MAPK通路促進(jìn)心肌肥厚。
[Abstract]:Aim: to investigate the effect of 尾 3 adrenoceptor (尾 3 AR) on myocardial hypertrophy in neonatal SD rats and its mechanism. Methods: neonatal SD rat cardiomyocytes were cultured in vitro. The cells were transfected with lentivirus carrying 尾 3-AR gene and induced by norepinephrine (NE) for 48h to establish cardiomyocyte hypertrophy model. The experiment was divided into four groups: control group (blank control group), NE group (myocardial hypertrophy group), NE group (尾 3-AR group) transfected with 尾 3-AR gene, and NE group (empty virus group) transfected with empty virus. The expression of green fluorescent protein (GFP) in viral transfection group was observed by reverse fluorescence microscope. Western blot was used to detect the expression of 尾 3-AR, p38 MAPK, ERK1 / 2, phosphorylated p38 MAPK and ERK (p-ERK1 / 2) and proto-oncogene c-mycc-fos. Results: (1) the expression of 尾 _ 3-AR gene was significantly higher than that of the blank control group. (2) the expression of proto-oncogene c-mycnc-fos was detected by Western blot, and the expression of c-mycfs in NE group, 尾 _ 3-AR group and empty virus group was higher than that in blank control group. The expression of c-fos in 尾 3-AR group was significantly higher than that in NE group. (3) the phosphorylation levels of p38 MAPK and ERK1 / 2 in NE group, 尾 3-AR group and empty virus group were significantly higher than those in control group, and the highest expression was found in 尾 3-AR group. Conclusion: lentivirus-mediated 尾 _ 3-AR gene transfection can effectively overexpression 尾 _ 3-AR. 尾 _ 3-AR may promote myocardial hypertrophy through MAPK pathway.
【作者單位】： 新疆石河子大學(xué)醫(yī)學(xué)院第一附屬醫(yī)院心內(nèi)二科;新疆醫(yī)科大學(xué)第一附屬醫(yī)院心臟中心;
【基金】：國(guó)家自然科學(xué)基金-地區(qū)科學(xué)基金項(xiàng)目(No:81260028) 石河子大學(xué)科學(xué)技術(shù)研究發(fā)展計(jì)劃“自然科學(xué)與技術(shù)創(chuàng)新”團(tuán)隊(duì)創(chuàng)新項(xiàng)目(No:2011ZRKXTD-07)
【分類號(hào)】：R541.6
【相似文獻(xiàn)】
相關(guān)期刊論文 前6條
1 樊守剛;吳小濤;王運(yùn)濤;李果;;骨髓間充質(zhì)干細(xì)胞經(jīng)慢病毒轉(zhuǎn)染綠色熒光蛋白后的體內(nèi)觀察[J];東南大學(xué)學(xué)報(bào)(醫(yī)學(xué)版);2010年02期
2 劉君;向川;衛(wèi)小春;;不同轉(zhuǎn)染方法對(duì)逆轉(zhuǎn)錄病毒轉(zhuǎn)染效率影響的研究[J];實(shí)用骨科雜志;2008年12期
3 王新武;劉文革;;人bFGF重組慢病毒轉(zhuǎn)染BMSCs的實(shí)驗(yàn)研究[J];中國(guó)現(xiàn)代醫(yī)生;2010年06期
4 方敏，，王立人，雷學(xué)鋒，畢愛(ài)華;逆轉(zhuǎn)錄病毒轉(zhuǎn)染的小鼠IL－3基因在NIH／3T3細(xì)胞中的表達(dá)[J];同濟(jì)醫(yī)科大學(xué)學(xué)報(bào);1996年06期
5 劉濤,李金蓮;大鼠三叉神經(jīng)脊束核尾側(cè)亞核淺層內(nèi)GABA樣、L-ENK樣、GlyT2樣陽(yáng)性末梢與Sindibis病毒轉(zhuǎn)染表達(dá)GFP神經(jīng)元之間的聯(lián)系[J];中國(guó)神經(jīng)科學(xué)雜志;2002年04期
6 ;[J];;年期
本文編號(hào)：2076083
本文鏈接：http://sikaile.net/kejilunwen/jiyingongcheng/2076083.html
