人脐带间充质干细胞对感染状态下人肺泡Ⅱ型上皮细胞的影响

发布时间：2018-06-25 07:03

本文选题：人脐带间充质干细胞 + 人肺泡Ⅱ型上皮细胞(A)　；参考：《中山大学学报(医学科学版)》2017年06期

【摘要】：【目的】探讨人脐带间充质干细胞(h UCMSC)对感染铜绿假单胞菌(PA)状态下人肺泡Ⅱ型上皮细胞(A549)的影响。【方法】人肺泡Ⅱ型上皮细胞A549(1×105/m L)2 m L与PA(3×104CFU/m L)2 m L共培育6 h后,加入h UCMSC(1×106/m L)2 m L为实验组,加入等量磷酸缓冲液(PBS)为感染组,A549与PBS及培养基共培育为对照组。比较组间A549细胞形态学改变(透射电镜),A549细胞存活率(CCK-8新型细胞增殖检测试剂盒),A549细胞凋亡率(Annexin V-FITC/PI双染色流式细胞仪)以及A549肺表面活性物质A(SP-A)表达量(Western Blot)。【结果】电镜细胞形态学观察显示,感染组A549细胞受损明显,胞质出现空泡变性,染色质高度凝集,可见凋亡小体;实验组细胞包膜结构完整,核膜完整,核仁明显,核染色质电子密度低,染色质均一,未见凋亡小体;对照组A549细胞结构完整,细胞膜表面微绒毛丰富,核膜完整,核周间隙结构正常,染色质均一;感染组与对照组相比,感染组A549细胞细胞存活率显著降低[(70.35±2.89)%与(97.37±2.07)%,n=3,P0.01],凋亡率显著增加[(8.63%±0.16)%与(2.55±0.11)%,n=3,P0.01],SP-A表达显著降低[(0.105±0.01)与(0.232±0.015),n=3,P0.01];实验组与感染组相比,实验组细胞存活率显著增加[(85.69±3.07)%与(70.35±2.89)%,n=3,P0.01],凋亡率显著下降[(6.34±0.12)%与(8.63±0.16)%,n=3,P0.01],感染组中,PA可能损伤A549细胞,使其SP-A表达量著减少(n=5,P0.05),实验组中,h UCMSC可能对感染后A549细胞的保护作用,其SP-A表达量增加(n=5,P0.05)。【结论】h UCMSC能抑制感染状态下的A549细胞凋亡,保护A549细胞分泌SP-A。
[Abstract]:[Objective] to investigate the effect of human umbilical cord mesenchymal stem cells (H UCMSC) on human alveolar type II epithelial cells (A549) in human infected Pseudomonas aeruginosa (PA). [Methods] the A549 (1 * 105/m L) 2 m L and PA (3 x 104CFU/m L) of human alveolar type II epithelial cells were incubated in 2, and 2 were added to the experimental group, adding equal amount of phosphoric acid. The buffer solution (PBS) was the infection group, and the A549 and PBS and the culture medium were co cultured as the control group. The morphological changes of A549 cells (transmission electron microscopy), the survival rate of A549 cells (CCK-8 new cell proliferation detection kit), the apoptosis rate of A549 cells (Annexin V-FITC/PI double dyed flow cytometry) and A (SP-A) expression of A549 lung surfactant were compared. N Blot). [results] the morphological observation of electron microscope showed that the A549 cells in the infected group were damaged obviously, the cytoplasm appeared vacuolated degeneration, the chromatin was highly agglutinated, and the apoptotic body was visible. The cell membrane structure of the experimental group was complete, the nuclear membrane was complete, the nucleolus was obvious, the nucleus chromatin electron density was low, the chromatin homogeneous, no apoptotic body was found, and the control group A549 cell junction was not found. The cell membrane surface microvilli was rich, the nuclear membrane was intact, the structure of the nuclear perinuclear space was normal, and the chromatin was uniform. Compared with the control group, the survival rate of A549 cells in the infection group was significantly lower [(70.35 + 2.89)% and (97.37 + 2.07)%, n=3, P0.01], and the apoptosis rate was significantly increased [(8.63% + 0.16)% and (2.55 + 0.11)%, n=3, P0.01], SP-A expression was significantly reduced. [(0.105 + 0.01) and (0.232 + 0.015), n=3, P0.01]; compared with the infection group, the cell survival rate of the experimental group was significantly increased [(85.69 + 3.07)% and (70.35 + 2.89)%, n=3, P0.01], and the apoptosis rate decreased significantly [(6.34 + 0.12)% and 8.63 + 0.16), n=3, P0.01], PA may damage A549 cells, making the SP-A expression reduction (n=5, P0.05), experiments In the group, H UCMSC may protect the infected A549 cells and increase the expression of SP-A (n=5, P0.05). [Conclusion] h UCMSC can inhibit the apoptosis of A549 cells in the infected state and protect the SP-A. secretion of A549 cells.
【作者单位】：广东省妇幼保健院新生儿科;广东省妇幼保健院检验科;
【基金】：广州市科技计划项目(2013J4100008)
【分类号】：R722.1

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