人羊膜间充质干细胞对松木屑烟雾溶液染毒大鼠肺泡Ⅱ型上皮细胞增殖、凋亡及炎症因子影响的研究  

Effects of human amniotic mesenchymal stem cells on proliferation,apoptosis and inflammatory factors of alveolar typeⅡepithelial cells in rats exposed to pine sawdust smoke solution

作　　者：朱秀连 崔培 詹球 李榕生 蒙凤姬 朱富军 杨福旺 童亚林 辛海明 ZHU Xiulian;CUI Pei;ZHAN Qiu;LI Rongsheng;MENG Fengji;ZHU Fujun;YANG Fuwang;TONG Yalin;XIN Haiming(Animal Laboratory,The 924th Hospital of the Joint Logistics Support Force of Chinese PLA,Guilin 541002,Guangxi Zhuang Autonomous Region,China;Burn Plastic Skin Beauty Center,The 924th Hospital of the Joint Logistics Support Force of Chinese PLA,Guilin 541002,Guangxi Zhuang Autonomous Region,China)

机构地区：[1]联勤保障部队第924医院动物实验室,广西桂林541002 [2]联勤保障部队第924医院烧伤整形皮肤美容中心,广西桂林541002

出　　处：《解放军医学院学报》2024年第5期528-534,共7页Academic Journal of Chinese PLA Medical School

基　　金：广西壮族自治区临床重点专科建设项目经费资助;广西科技基地与人才专项(桂科AD18126016);桂林市自筹经费科技项目(20210102z)。

摘　　要：背景烟雾吸入性肺损伤在烧伤患者中较为常见,吸入的有毒气体、颗粒等易引起急性肺损伤(acute lung injury,ALI),目前无特异性治疗方法。目的研究人羊膜间充质干细胞(human amniotic mesenchymal stem cells,hAMSCs)对松木屑烟雾溶液染毒大鼠肺泡Ⅱ型上皮细胞(alveolar typeⅡepithelial cells,AT-Ⅱ)增殖、凋亡及炎症因子的影响。方法采用酶消化法分离培养hAMSCs和AT-Ⅱ,采用流式细胞术和免疫荧光分别鉴定hAMSCs和AT-Ⅱ;松木屑烟雾溶液染毒AT-Ⅱ复制烟雾诱导的细胞损伤。实验细胞分为对照组(无血清DMEM/F12)、致伤组(0.75‰松木屑烟雾溶液染毒)和治疗组(0.75‰松木屑烟雾溶液染毒,hAMSCs与AT-Ⅱ共培养)。采用CCK-8检测细胞的增殖活性;Annexin V-FITC/PI染色检测细胞凋亡;Elisa检测炎症因子肿瘤坏死因子α(tumor necrosis factor-α,TNF-α)、白细胞介素(interleukin,IL)-6和IL-10的表达。结果与对照组比较,致伤组AT-Ⅱ的活性降低(P<0.01),经hAMSCs治疗后,与致伤组比较,AT-Ⅱ细胞活性增加(P<0.01);致伤组细胞的凋亡率在12 h和24 h均增加(P<0.01),hAMSCs治疗后细胞凋亡率降低(P<0.01);细胞致伤后炎症因子TNF-α、IL-6和IL-10的含量增加(P<0.05),经hAMSCs共培养12 h和24 h后,促炎因子TNF-α和IL-6的表达均下调(P<0.05),IL-10含量增加(P<0.05)。结论hAMSCs可以促进松木屑烟雾溶液诱导的AT-Ⅱ增殖和抑制细胞凋亡,可能与调节炎症因子的表达有关,为探究hAMSCs治疗烟雾引起的急性肺损伤提供理论依据。Background Smoke inhalation lung injury is common in burn patients.Inhaled toxic gases and particles are easy to cause acute lung injury(ALI),and there is no specific treatment at present.Objective To investigate the effects of human amniotic mesenchymal stem cells(hAMSCs)on proliferation,apoptosis and inflammatory factors of alveolar typeⅡepithelial cells(AT-Ⅱ)in rats exposed to pine sawdust smoke solution.Methods hAMSCs and AT-Ⅱwere isolated and cultured by enzyme digestion,and identified by flow cytometry and immunofluorescence,respectively;Cell damage induced by AT-Ⅱreplication smoke was exposed to pine sawdust smoke solution.The cells were divided into control group(serum-free DMEM/F12),injury group(exposure to 0.75‰pine sawdust smoke solution)and treatment group(exposure to 0.75‰pine sawdust smoke solution and hAMSCs cocultured with AT-Ⅱ).CCK-8 assay was used to detect the proliferation of cells;Annexin V-FITC/PI staining was used to detect apoptosis;Elisa was used to analyze the expression of TNF-α,IL-6 and IL-10.Results Compared with the control group,the activity of AT-Ⅱin the injured group decreased(P<0.01),and after hAMSCs treatment,the activity of AT-Ⅱcells increased compared with the injury group(P<0.01);The apoptosis rate in the injured group increased at 12 h and 24 h(P<0.01),while decreased after hAMSCs treatment(P<0.01);The contents of inflammatory factors TNF-α,IL-6 and IL-10 increased after cell injury(P<0.05),and the expression levels of pro-inflammatory factors TNF-αand IL-6 decreased(P<0.05)after 12 h and 24 h of co-culture with hAMSCs,and the content of IL-10 increased after co-cultured for 12 h and 24 h(P<0.05).Conclusion hAMSCs can promote the proliferation and inhibit apoptosis of AT-Ⅱinduced by pine sawdust smoke solution,which may be related to regulating the expression of inflammatory factors,and provide theoretical reference for exploring the treatment of acute lung injury caused by smoke with hAMSCs.

关 键 词：人羊膜间充质干细胞 肺泡Ⅱ型上皮细胞 增殖 凋亡 炎症因子 

分 类 号：R563[医药卫生—呼吸系统]