姜黄素对脂多糖诱导的滋养细胞系HTR-8/SVneo中微小RNA-155表达及细胞凋亡、侵袭的影响  

The effect of curcumin on the expression of miR-155 to apoptosis and invasion of extravillus trophoblast cells treated by lipopolysaccharide

作　　者：洪国敏 龚萍[2] 杨沐怿 沈莉[2] 刁振宇[2] 胡娅莉[2] 

机构地区：[1]南京中医药大学中西医结合鼓楼临床医学院,210023 [2]南京大学医学院附属鼓楼医院妇产科

出　　处：《中华妇产科杂志》2016年第11期859-864,共6页Chinese Journal of Obstetrics and Gynecology

基　　金：国家自然科学基金（81370724、81571462）

摘　　要：目的探讨姜黄素对脂多糖（LPS）诱导的滋养细胞系HTR-8/SVneo细胞中微小RNA-155（miR-155）表达、细胞凋亡和侵袭的影响。方法体外培养滋养细胞系HTR-8/SVneo细胞。实验共4组：姜黄素＋LPS组（姜黄素浓度分别为12.5、25、50μmol/L预处理后，加入100 ng/ml LPS）、LPS组（加入100 ng/ml LPS）、重组腺病毒组（加入Ad-miR-155，感染复数为100）和空白对照组。采用实时荧光定量PCR、荧光素酶报告基因活性检测法和蛋白印迹法检测姜黄素＋LPS组、LPS组和空白对照组细胞的miR-155表达水平、荧光素酶转录活性及核因子κB（NF-κB）蛋白的表达水平；细胞凋亡酶联免疫法和细胞侵袭试验检测姜黄素＋LPS组、LPS组、重组腺病毒组和空白对照组细胞凋亡和侵袭能力的情况。结果（1）实时荧光定量PCR技术结果显示，LPS组HTR-8/SVneo细胞中miR-155的表达水平为空白对照组的（2.13±0.22）倍，两组比较，差异具有统计学意义（P〈0.01）；12.5、25、50μmol/L姜黄素＋LPS组HTR-8/SVneo细胞中miR-155的表达水平分别为LPS组的（0.37±0.08）、（0.68±0.14）、（0.49±0.09）倍，分别比较，差异均有统计学意义（P〈0.05）。（2）荧光素酶报告基因活性检测法显示， LPS组荧光素酶的转录活性为空白对照组的（2.25±0.56）倍，12.5、25、50μmol/L姜黄素＋LPS组荧光素酶转录活性分别为LPS组的（0.80±0.07）、（0.74±0.05）、（0.49±0.19）倍，差异均有统计学意义（P均〈0.05）。（3）蛋白印迹法检测显示，LPS组HTR-8/SVneo细胞中p65蛋白的表达水平为1.50±0.22，明显高于空白对照组（0.95±0.25，P〈0.01）。不同浓度姜黄素＋LPS组，姜黄素浓度为12.5μmol/L时，p65蛋白表达水平为0.31±0.07，与LPS组相比，下调最明显（P〈0.01）；浓度为25、50μmol/L时，蛋白表达水平分别为0.75±0.14、0.49±0.08，分别与LPS组比较，差异均有统计学意义（P〈0.Objective To investigate the effect of curcumin on the expression of miR-155, apoptosis and invasion of extravillus trophoblast cells treated by lipopolysaccharide （LPS）. Methods Human trophoblast cells （HTR-8/SVneo cells） were divided into 4 groups, the curcumin ＋ LPS group （pre-treated by curcumin of 12.5, 25, 50 μmol/L, then LPS of 100 ng/ml）, the LPS group （100 ng/ml）, the recombinant adenovirus group（miR-155,multiplicity of infection100）and the control group. The miR-155 level in HTR-8/SVneo cells was measured by real-time PCR, and the expression of NF-κB was analyzed by luciferase gene expression. The apoptosis of HTR-8/SVneo cells was tested by cell death detection ELISA and the level of NF-κB in HTR-8/SVneo cells was measured by western blot. In addition, transwell was used to test the invasive ability of HTR-8/SVneo cells in all the groups. Results （1） The intracellular expression of miR-155 in the LPS group was （2.13 ± 0.22） times of the control group （P〈0.01）; and the expressions of miR-155 in 12.5,25,50μmol/L curcumin＋LPS groups were（0.37±0.08）,（0.68±0.14）,（0.49±0.09）times as the LPS group, with statistically significant difference （P〈0.05）. （2） The expreesion of NF-κB in the LPS group was（2.25 ± 0.56）times of the control group. The expreesion of NF-κB in the 12.5,25,50 μmol/L curcumin＋LPS groups were（0.80 ± 0.07）,（0.74 ± 0.05）,（0.49 ± 0.19） times to the LPS group, with statistically significant difference（P〈0.01）. （3）The p65 protein in the LPS group was（1.50 ± 0.22）, significantly higher than the control group（0.95 ± 0.25,P〈0.01）. In 12.5,25,50 μmol/L curcumin＋LPS groups, the p65 protein were（0.31 ± 0.07）,（0.75 ± 0.14）,（0.49 ± 0.08）. Compared with the LPS group, p65 was down-regulated by curcumin, with statistically significant difference （P〈0.05）. （4） In the cell death detection ELISA, the A value in the control group, the LPS group and the recombined adenovi

关 键 词：先兆子痫 滋养层 姜黄素 细胞凋亡 

分 类 号：R714.244[医药卫生—妇产科学]