右美托咪定通过lncRNA H19介导香烟提取物诱导的肺泡巨噬细胞效应研究  

The cellular effects of dexmedetomidine in cigarette smoke extract-induced macrophages via lncRNA H19

作　　者：夏延贞 王龙珍 莫靓 商雄跃 XIA Yan-zhen;WANG Long-zhen;MO Jing;SHANG Xiong-yue(Department of critical medicine,Pukou branch,Jiangsu Peopled Hospital,Nanjing Jiangsu 211800,China)

机构地区：[1]江苏省人民医院浦口分院重症医学科,江苏南京211800

出　　处：《毒理学杂志》2021年第1期56-60,66,共6页Journal of Toxicology

摘　　要：目的探索右美托咪定(Dexmedetomidine,DEX)通过长链非编码RNA(long noncoding RNA,lncRNA)H19对香烟烟雾提取物(cigarette smoke extract,CSE)诱导的RAW264.7巨噬细胞活力、凋亡和自噬的影响。方法制备CSE及CSE干预巨噬细胞模型,运用体外细胞培养,DEX以不同浓度(2、4和8μmol/L)作用于CSE诱导的巨噬细胞,采用MTT法检测细胞存活率,流式细胞术检测模型细胞的凋亡率,采用蛋白质印迹法(Western blot)检测细胞中凋亡相关蛋白Bax和Bcl-2及自噬相关蛋白LC3Ⅱ和ATG7表达情况,实时荧光定量PCR(qRT-PCR)检测H19表达水平;为了进一步验证DEX对CSE诱导的巨噬细胞增殖、凋亡和自噬影响,运用双酶切法构建pcDNA3.1-H19表达载体。结果一定剂量的DEX可明显拮抗CSE诱导的巨噬细胞活力下降作用(P<0.05),具有浓度依赖性;DEX抑制CSE诱导的巨噬细胞凋亡,且细胞中凋亡蛋白Bax明显下调(P<0.05),抗凋亡蛋白Bcl-2上调(P<0.05),自噬相关蛋白LC3Ⅱ和ATG7表达上调(P<0.05),H19表达下调(P<0.05);成功构建pcDNA3.1-H19表达载体,DEX作用于CSE诱导的巨噬细胞,细胞活力明显降低(P<0.05),且细胞中凋亡蛋白Bax上调(P<0.05),抗凋亡蛋白Bcl-2下调(P<0.05),自噬相关蛋白LC3Ⅱ和ATG7表达下调(P<0.05)。结论DEX抑制H19表达,增强CSE诱导的巨噬细胞活力,抑制细胞凋亡和自噬。Objective To investigate the effects of dexmedetomidine(DEX)on the viability,apoptosis and autophagy of PAW264.7 macrophages induced by cigarette smoke extract(CSE)through long noncoding RNA(lncRNA)H19.Methods CSE and CSE intervention model of macrophages were prepared.Cells were cultured in vitro,and DEX was applied to CSE-induced macrophages at different concentrations(2,4 and 8μmol/L).MTT method was used to detected the cell survival rate,and the apoptosis rate of model cells was detected by flow cytometry.Western blot was used to detected the expressions of apoptosis-related proteins Bax,Bcl-2,autophagy-related proteins LC3Ⅱand ATG7 in cells.Real-time fluorescence quantitative PCR(qRT-PCR)was used to detect the expression level of H19;in order to further verify the effect of DEX on CSE-induced macrophage proliferation and apoptosis,a double enzyme digestion method was used to construct the pcDNA3.1-H19 expression vector.Results DEX can significantly promote the proliferation of CSE-induced macrophages(P<0.05),with concentration-dependent mannar;the apoptosis of CSE-induced macrophages was inhibited by DEX,and the apoptosis protein Bax in the cells was significantly down-regulated(P<0.05),anti-apoptosis protein Bcl-2 was significantly up-regulated(P<0.05),the expressions of autophagy-related proteins LC3Ⅱand ATG7 were significantly up-regulated(P<0.05),and the expression of H19 was significantly down-regulated(P<0.05);the pcDNA3.1-H19 expression vector was successfully constructed,while CSE-induced macrophages were treated by DEX,the cell viability of the cells was significantly reduced(P<0.05),and the apoptosis protein Bax was significantly up-regulated(P<0.05),and the anti-apoptotic protein Bcl-2 was significantly down-regulated(P<0.05).The expressions of phage-related proteins LC3Ⅱand ATG7 were significantly down-regulated(P<0.05).Conclusion DEX can inhibit expression of H19,enhance CSE-induced macrophage viability,and inhibit its apoptosis and autophagy.

关 键 词：右美托咪定 lncRNA H19 香烟烟雾提取物 凋亡 自噬 

分 类 号：R614[医药卫生—麻醉学] R99[医药卫生—外科学]