铜绿假单胞菌感染后人脐带间充质干细胞对A549细胞凋亡的保护作用研究
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摘要
目的观察感染状态下应用人脐带间充质干细胞(hUCMSCs)对A549细胞凋亡起到的保护作用。方法无菌条件下从胎儿脐带和壁蜕膜组织中采集hUCMSCs标本,接种铜绿假单胞菌(PA)于A549细胞以建立A549细胞损伤模型,分析PA对A549细胞凋亡的影响及hUCMSCs对细胞凋亡的保护。结果经PA感染后,导致A549细胞生长缓慢、老化,细胞空泡化;感染状态下A549细胞SP-A表达水平下降,与未感染A549细胞相比差异明显(t=7.745,P=0.001);感染A549细胞接种hUCMSCs后,SP-A表达水平上调,与感染组相比差异明显(t=3.872,P=0.017);PA感染后A549细胞凋亡率加快,存活率降低,接种hUCMSCs后A549细胞凋亡率下降,存活率上调。结论 A549细胞在PA感染状态下通过接种hUCMSCs,可起到修复细胞损伤,抑制细胞凋亡的作用。
OBJECTIVE To observe the protective effect of human umbilical cord mesenchymal stem cells(hUCMSCs)on apoptosis of A549 cells of patients with infections.METHODS The hUCMSCs specimens were collected from fetal umbilical cord and decidua tissues under aseptic condition,the A549 cells were inoculated with Pseudomonas aeruginosa so as to establish A549 cell damage model,and the impact of P.aeruginosa on the apoptosis of the A549 cells and the protective effect of hUCMSCs on the apoptosis were observed.RESULTS The P.aeruginosa infection resulted in the slow growth,aging and vacuolation of the A549 cells,the expression of SP-A in the A549 cells was reduced under the infection state,as compared with the A549 cells that were not infected,there was significant difference(t=7.745,P=0.001).The expression of SP-A in the A549 cells was up-regulated after the inoculation of hUCMSCs,as compared with infection group,there was significant difference(t=3.872,P=0.017).The rate of apoptosis of the A549 cells accelerated after the P.aeruginosainfection,and the survival rate was reduced;the rate of apoptosis of the A549 cells was reduced after the inoculation of hUCMSCs,and the survival rate was up-regulated.CONCLUSIONThe inoculation of hUCMSCs may repair the damage of the A549 cells infected with P.aeruginosainfection and inhibit the apoptosis.
OBJECTIVE To observe the protective effect of human umbilical cord mesenchymal stem cells(hUCMSCs)on apoptosis of A549 cells of patients with infections.METHODS The hUCMSCs specimens were collected from fetal umbilical cord and decidua tissues under aseptic condition,the A549 cells were inoculated with Pseudomonas aeruginosa so as to establish A549 cell damage model,and the impact of P.aeruginosa on the apoptosis of the A549 cells and the protective effect of hUCMSCs on the apoptosis were observed.RESULTS The P.aeruginosa infection resulted in the slow growth,aging and vacuolation of the A549 cells,the expression of SP-A in the A549 cells was reduced under the infection state,as compared with the A549 cells that were not infected,there was significant difference(t=7.745,P=0.001).The expression of SP-A in the A549 cells was up-regulated after the inoculation of hUCMSCs,as compared with infection group,there was significant difference(t=3.872,P=0.017).The rate of apoptosis of the A549 cells accelerated after the P.aeruginosainfection,and the survival rate was reduced;the rate of apoptosis of the A549 cells was reduced after the inoculation of hUCMSCs,and the survival rate was up-regulated.CONCLUSIONThe inoculation of hUCMSCs may repair the damage of the A549 cells infected with P.aeruginosainfection and inhibit the apoptosis.
引文
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[13]刘晓阳,任雁宏,詹庆元.机械牵张诱发肺泡Ⅱ型上皮细胞黏附相关的抗损伤机制研究[J].山西医药杂志,2015(1):3-6.
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[15]Yang LM,Liu Y,Zhao J,et al.Characterization of human umbilical cord mesenchymal stem cells following tissue mass culture[J].Cell Mol Biol,2014,60(1):12-18.
[2]周志刚,李志忠,林永新,等.人脐带间充质干细胞定向诱导分化成类神经元的实验研究[J].中国病理生理杂志,2015,31(2):229-233.
[3]Lim JY,Chang HJ,Jin AJ,et al.Therapeutic effects of human umbilical cord blood-derived mesenchymal stem cells after intrathecal administration by lumbar puncture in a rat model of cerebral ischemia[J].Stem Cell Res Ther,2011,2(5):38.
[4]Shaer A,Azarpira N,Aghdaie MH,et al.Isolation and characterization of human mesenchymal stromal cells derived from placental decidua basalis;umbilical cord Wharton's jelly and amniotic membrane[J].Pak J Med Sci,2014,30(5):1022-1026.
[5]惠玲,刘毅,宋枚.绿色荧光蛋白和胰岛素基因共表达重组腺病毒载体的构建及感染人脐带间充质干细胞的实验研究[J].解放军医药杂志,2013,25(10):6-10.
[6]Seo DK,Kim JH,Min J,et al.Enhanced axonal regeneration by transplanted Wnt3a-secreting human mesenchymal stem cells in a rat model of spinal cord injury[J].Acta Neurochir(Wien),2017,159(5):947-957.
[7]肖恒,赵成岭,陈诗军,等.脐带间充质干细胞恶性分化并促进肺癌A549细胞移植瘤生长及转移[J].中国组织化学与细胞化学杂志,2017,26(4):342-348.
[8]Stanko P,Kaiserova K,Altanerova V,et al.Comparison of human mesenchymal stem cells derived from dental pulp,bone marrow,adipose tissue,and umbilical cord tissue by gene expression[J].Biomed Pap Med Fac Univ Palacky Olomouc Czech Repub,2014,158(3):373-377.
[9]裴宇,刘晓程,姜亦瑶,等.不同低氧浓度预处理对人脐带间充质干细胞抗凋亡能力的影响[J].中华全科医学,2015,13(8):1247-1249.
[10]段辉.人脐带间充质干细胞的条件培养基对炎症因子致人脐静脉内皮细胞凋亡的保护作用[J].实用医学杂志,2016,32(22):3645-3649.
[11]黄一,王婷婷,游凯,等.NIX介导的细胞自噬在低氧诱导ATⅡ凋亡中的作用[J].西南国防医药,2015,25(8):847-850.
[12]张春春,轩亚茹,王亚华,等.紫花牡荆素对肺癌A549细胞增殖及凋亡的影响[J].山东医药,2016,56(36):13-16.
[13]刘晓阳,任雁宏,詹庆元.机械牵张诱发肺泡Ⅱ型上皮细胞黏附相关的抗损伤机制研究[J].山西医药杂志,2015(1):3-6.
[14]Dapat C,Saito R,Suzuki H,et al.Quantitative phosphoproteomic analysis of host responses in human lung epithelial(A549)cells during influenza virus infection[J].Virus Res,2014,179:53-63.
[15]Yang LM,Liu Y,Zhao J,et al.Characterization of human umbilical cord mesenchymal stem cells following tissue mass culture[J].Cell Mol Biol,2014,60(1):12-18.