绵羊肺炎支原体对人肺泡Ⅱ型上皮细胞活性及炎症反应的影响

Effect of Mycoplasma ovipneumoniae on the activity and inflammatory response of human alveolar typeⅡepithelial cells

为探究绵羊肺炎支原体(MO)是否对人体造成潜在影响,本研究以人肺泡Ⅱ型上皮细胞为细胞模型,将MO Y98株荧光染色后感染人肺泡Ⅱ型上皮细胞,于激光共聚焦显微镜下可见MO存在于细胞中;分别利用CCK-8及CellTiter-LumiTM试剂盒检测细胞的增殖及活力,结果显示MO对人肺泡Ⅱ型上皮细胞的增殖及活性均无显著影响;扫描电镜检测细胞形态,结果显示肺泡上皮细胞伪足明显增多;基因芯片分析筛选出3个mRNA及5个lncRNA,荧光定量PCR验证后确定SCNN1G、LOX、ST6GALNAC1及lncRNA RP11-29G8.3-001在MO感染后的转录水平发生显著变化;荧光定量PCR检测肺泡上皮细胞中IL-1β、TNF-α、IFN、IRF7等炎症因子,结果显示这些因子基因的转录水平无明显变化;采用将lncRNA RP11-29G8.3-001过表达的方法对其功能进行研究,结果显示少量lncRNA RP11-29G8.3-001表达增高不会调控细胞免疫相关基因的表达。本研究结果表明MO并不降低人肺泡Ⅱ型上皮细胞的增殖及活性,且MO对人肺泡Ⅱ型上皮细胞炎症因子的表达无显著影响,提示MO对人肺泡Ⅱ型上皮细胞影响较小。本研究为防范MO对人呼吸系统存在的潜在影响提供了实验依据。

To explore whether Mycoplasma ovipneumoniae(MO)could cause potential harm to the human body,human alveolar typeⅡepithelial cells were selected as an infection model.Human alveolar typeⅡepithelial cells infected with MO Y98 were stained and MO can be observed under laser confocal microscopy.Cell viability detection results revealed that Mycoplasma ovipneumoniae had no significant effect on the proliferation and activity of human alveolar epithelial cells.Scanning electron microscopy detection of cell morphology showed that pseudopodia of the cells increased significantly.Microarray assay of human alveolar epithelial cells infected with Mycoplasma ovipneumoniae indicated that SCNN1G,LOX,ST6GALNAC1 and lncRNA RP11-29G8.3-001 changed significantly,but no significant change was observed of inflammatory factors,including IL-1β,TNF-α,IFN and IRF7.Moreover,overexpression of the lncRNA RP11-29G8.3-001 showed that slightly increased expression of the lncRNA could not regulate cell immune-related genes.In conclusion,we found that Mycoplasma ovipneumoniae could not affect the proliferation and activity of human alveolar epithelial cells.Also,infection of Mycoplasma ovipneumoniae had no significant effect on the expression of inflammatory factors,suggesting that Mycoplasma ovipneumoniae could not cause damage to human alveolar typeⅡepithelial cells.This study provides an experimental basis for preventing the potential harm of Mycoplasma ovipneumoniae to human respiratory system.