摘要
目的 分析抑制Toll样受体4(TLR4)基因对TORCH感染的人胎盘滋养细胞生物学行为的影响及机制。方法 将人胎盘滋养细胞培养后分为对照组、TORCH感染组、TLR4过表达组、TLR4沉默组四组。对照组不做任何处理,TORCH感染组为弓形虫病毒(Tox)感染人胎盘滋养细胞,TLR4过表达组为TORCH感染人胎盘滋养细胞+TLR4过表达质粒转染,TLR4沉默组为TORCH感染人胎盘滋养细胞+TLR4沉默质粒转染。分析TLR4基因对TORCH感染人胎盘滋养细胞增殖抑制、凋亡、细胞周期分布的影响。采用免疫印迹法(WB)测定细胞Wnt/β-连环蛋白通路蛋白表达量。结果 与TLR4过表达组比较,TLR4沉默组细胞增殖抑制率及细胞凋亡率均降低(P<0.05);G1期细胞数降低,S期、G2/M期细胞数升高(P<0.05);Wnt、β-catenin、Bcl-2、CyclinD1蛋白表达量升高,Bax蛋白表达量降低(P<0.05)。结论 抑制TLR4基因表达可抑制TORCH感染的人胎盘滋养细胞凋亡,促进增殖,调控周期分布,其作用机制可能与激活Wnt/β-catenin通路相关。
OBJECTIVE To observe the influence of inhibition of Toll like receptor 4(TLR4) gene on biological behavior of human placental trophoblast cells of patients with TORCH infection and its mechanisms. METHODS The human placental trophoblast cells were divided into the control group, the TORCH infection group, the TLR4 overexpression group and the TLR4 silence group after the culture. The human placental trophoblast cells of the control group were not given any treatment, the human placental trophoblast cells of the TORCH infection group were infected with Toxoplasma(Toxoplasma, Tox), the human placental trophoblast cells of the TLR4 overexpression group were treated with TORCH infection plus transfection of TLR4 overexpression plasmid, and the human placental trophoblast cells of the TLR4 silence group were treated with TORCH infection plus transfection of TLR4 silencing plasmid. The influence of TLR4 gene on inhibition of proliferation, apoptosis and cell cycles of the human placental trophoblast cells with TORCH infection was observed. The expression level of Wnt/β-catenin pathway protein in the cells was determined by using Western Blot method. RESULTS The rates of inhibition of proliferation and apoptosis of the cells were significantly lower in the TLR4 silence group than in the TLR4 overexpression group(P<0.05);the counts of cells were reduced in G1 phase, while the counts of cells were significantly increased in S phase, G2/M phase(P<0.05). The expression levels of Wnt, β-catenin, B cell lymphoma-2(Bcl-2) and Cyclin D1 proteins were significantly elevated, while the expression level of BCL2-associated X protein(Bax) was significantly reduced(P<0.05). CONCLUSION The inhibition of TLR4 gene may inhibit the apoptosis of the human placental trophoblast cells with TORCH infection, promote the proliferation and regulate the distribution of cycle, and the mechanisms may be associated with the activation of Wnt/β-catenin pathway.
作者
陈秀芸
赵洁
亓淑芳
时美
巩雯雯
CHEN Xiu-yun;ZHAO Jie;QI Shu-fang;SHI Mei;GONG Wen-wen(Jinan City Second Maternal and Child Health Hospital,Jinan,Shandong 271199,China)
出处
《中华医院感染学杂志》
CAS
CSCD
北大核心
2022年第1期115-119,共5页
Chinese Journal of Nosocomiology
基金
山东省医药卫生科技发展计划项目(2017WSSA15107)。
