本文選題：結核分枝桿菌 + Mcl-基因　； 參考：《中國病理生理雜志》2015年12期

【摘要】：目的:通過RNA干擾技術特異性沉默Mcl-1基因,探討下調Mcl-1基因對感染不同毒力結核桿菌小鼠腹腔巨噬細胞凋亡的影響。方法:分別用制備好的新疆地區(qū)流行的優(yōu)勢強毒結核分枝桿菌臨床分離株(簡稱強毒株)、結核分枝桿菌國際標準強毒株H37Rv(簡稱H37Rv)、結核分枝桿菌國際標準無毒株H37Ra(簡稱H37Ra)和卡介苗(BCG)菌懸液感染BALB/c小鼠,再以篩選并構建好的Mcl-1-shRNA作用于感染的小鼠模型,同時設立相應對照組,于作用后1 d、3 d、5 d和7 d提取小鼠腹腔巨噬細胞,應用實時熒光定量PCR和Western blot檢測各組小鼠腹腔巨噬細胞中Mcl-1 mRNA和蛋白的表達;應用流式細胞術檢測各組巨噬細胞的凋亡水平。結果:小鼠被不同毒力的結核桿菌感染后其腹腔巨噬細胞中Mcl-1 mRNA和蛋白的表達水平均有不同程度的升高,其中以感染了強毒株和H37Rv的腹腔巨噬細胞升高最為明顯(P0.05);應用RNA干擾技術沉默Mcl-1基因后,Mcl-1mRNA和蛋白的表達水平明顯低于對照組(P0.05);流式細胞術分析顯示,下調Mcl-1基因的表達可誘導小鼠腹腔巨噬細胞凋亡。結論:應用Mcl-1-shRNA可有效沉默Mcl-1在感染了不同毒力結核桿菌小鼠腹腔巨噬細胞中的表達,并能上調巨噬細胞的凋亡水平。
[Abstract]:Aim: to study the effect of down-regulating Mcl-1 gene on peritoneal macrophage apoptosis in mice infected with different virulence Mycobacterium tuberculosis by RNA interference technique.Methods: the clinical isolates of dominant virulent Mycobacterium tuberculosis (virulent strain), virulent strain H37Rv1 (International Standard strain H37Rvv) and International Standard strain H37Ra( no strain of Mycobacterium tuberculosis) were prepared in Xinjiang, respectively.H37Raand Bacillus Calmette-Guerin Bacillus Calmette-Guerin (BCG) suspension infected BALB/c mice,Then Mcl-1-shRNA was used to screen and construct the infected mouse model, and the corresponding control group was set up. The peritoneal macrophages were extracted from mice on the 1st day, 3rd day, 5th day, and 7th day, respectively.The expression of Mcl-1 mRNA and protein in peritoneal macrophages was detected by real-time fluorescence quantitative PCR and Western blot, and the apoptotic level of macrophages in each group was detected by flow cytometry.Results: the expression levels of Mcl-1 mRNA and protein in peritoneal macrophages of mice infected with different virulence Mycobacterium tuberculosis were increased in varying degrees.Among them, the peritoneal macrophages infected with virulent strain and H37Rv increased the most significantly (P 0.05), the expression level of mcl-1 mRNA and protein in Mcl-1 gene silenced by RNA interference technique was significantly lower than that in control group (P 0.05). Flow cytometry analysis showed that the expression level of Mcl-1 mRNA and protein was significantly lower than that of control group.Down-regulating the expression of Mcl-1 gene can induce mouse peritoneal macrophage apoptosis.Conclusion: Mcl-1-shRNA can effectively silence the expression of Mcl-1 in the peritoneal macrophages of mice infected with different virulent Mycobacterium tuberculosis and can up-regulate the apoptosis level of macrophages.
【作者單位】： 石河子大學醫(yī)學院病理生理學教研室;石河子大學醫(yī)學院新疆地方與民族高發(fā)病教育部重點實驗室;石河子大學醫(yī)學院第一附屬醫(yī)院泌尿外科;石河子大學醫(yī)學院病原生物學與免疫學教研室;
【基金】：國家自然科學基金資助項目(No.81260241)
【分類號】：R378.911

【參考文獻】

相關期刊論文 前2條

1 董江濤;徐芳;田璽擇;姚楠;吳芳;章樂;王遠志;曹旭東;張萬江;;不同毒力結核分枝桿菌感染小鼠對肺泡巨噬細胞的凋亡率及其時相性變化的影響[J];中國免疫學雜志;2012年05期

2 張榮環(huán);趙菊;;髓細胞白血病因子-1研究進展[J];現代中西醫(yī)結合雜志;2014年06期

【共引文獻】

相關期刊論文 前10條

1 楊德芳;姚美玉;孫可豐;王欣欣;吉海榮;;Bcl-2、Bax和Mcl-1在異位子宮內膜中的表達[J];黑龍江醫(yī)藥科學;2014年04期

2 朱坤生;易敏;周向東;李升錦;;結核分枝桿菌eis基因表達對人THP-1細胞細胞因子分泌的影響[J];免疫學雜志;2015年08期

3 庹清章;董江濤;田璽擇;劉云霞;董偉杰;劉丹霞;李微;吳芳;章樂;張萬江;;結核分枝桿菌小分子熱休克蛋白Hsp16.3基因缺失突變菌株對感染小鼠肺泡巨噬細胞凋亡率的影響及其時相性變化[J];石河子大學學報(自然科學版);2013年01期

4 劉云霞;董偉杰;庹清章;劉丹霞;李微;張春軍;張輝;吳芳;章樂;張萬江;;結核分枝桿菌Hsp16.3對感染巨噬細胞凋亡的影響[J];石河子大學學報(自然科學版);2013年05期

5 于翔;陸佳嘉;王秦;于曉Y，

本文編號：1771780