摘要
目的探究过表达热休克蛋白60(HSP60)对骨髓间充质干细胞(MSCs)活性的影响及对光气急性肺损伤大鼠的疗效改变。方法利用腺病毒为载体转染HSP60至MSCs。用免疫印迹试验(Western blot)测量细胞转染前后HSP60的表达。利用CCK-8实验检测MSCs活力变化,流式凋亡实验检测MSCs凋亡能力的改变,Transwell实验观察MSCs迁移能力的改变。将60只SPF级雄性SD大鼠随机分为对照组、光气染毒组(吸入光气5min)、MSCs组(光气染毒,MSCs治疗组)和转染MSCs组(光气染毒,过表达HSP60 MSCs治疗组),每组各15只。通过肺部病理切片观察大鼠肺部病理改变,肺湿干比观察肺部水肿程度改变,检测肺泡灌洗液总细胞计数、总蛋白量及肺泡灌洗液和血清肿瘤坏死因子-α(TNF-α)和白细胞介素-6(IL-6)反映炎症变化。用Graphpad Prism 8.0软件对数据进行分析,两两比较采用非配对t检验,组间比较采用单因素方差分析。结果转染HSP60后,与未转染HSP60的MSCs[A=(0.27±0.02)]比较,转染HSP60的MSCs增殖能力[A=(0.69±0.05)]提高,差异有统计学意义(P<0.05)。与光气染毒组比较,MSCs组和转染后MSCs组大鼠肺组织水肿均有所减轻,炎症因子浸润减少。但转染后MSCs组较MSCs组肺部水肿减轻程度改善较为明显、IL-6、TNF-α下降程度更高,肺泡灌洗液总蛋白量及总细胞数更少,差异均有统计学意义(P<0.05)。结论 MSCs转染HSP60后,增强了增殖能力、抗凋亡能力、迁移能力及在光气急性肺损伤大鼠中的疗效。
Objective To investigate the effect of heat shock protein 60(HSP60)overexpression on the ability of bone marrow mesenchymal stem cells(MSCs)and its therapeutic effect on rats with phosgene induced acute lung injury.Methods HSP60 was transfected into MSCs by adenovirus.Western blot was used to measure the expressions of HSP60 before and after transfection.CCK-8 assay was used to detect the activity of MSCs,flow cytometry was used to detect the apoptotic ability of MSCs,and Transwell assay was used to observe the migration ability of MSCs.Sixty SPF grade male SD rats were randomly divided into control group,phosgene exposure group(inhalation of phosgene for 5 min),MSCs group(phosgene exposure,MSCs treatment group)and transfected MSCs group(phosgene exposure,overexpression of HSP60 MSCs treatment group).The pathological changes of lung were observed by lung pathological section,lung wet dry ratio,the degree of pulmonary edema,the total cell count and total protein content of alveolar lavage fluid,the inflammatory changes of tumor necrosis factor-α(TNF-α)and interleukin-6(IL-6)in BALF and serum were observed.The data were analyzed by Graphpad Prism 8.0 software.Paired comparisons were performed by non paired t-test.One way ANOVA was used for comparison between groups.Results The proliferation ability of MSCs transfected with HSP60[A=(0.69±0.05)]was significantly higher than that of MSCs not transfected with HSP60[A=(0.27±0.02)](P<0.05).Compared with the phosgene exposure group,the pulmonary edema and inflammatory factor infiltration of MSCs group and MSCs transfected group were reduced.However,compared with MSCs group,the degree of pulmonary edema in MSCs transfected group was significantly improved,the levels of inflammatory factors IL-6 and TNF-αwere significantly decreased,and the total protein content and total cell count in bronchoalveolar lavage fluid were less(P<0.05).Conclusion MSCs transfected with HSP60 can enhance the ability of proliferation,anti apoptosis,migration and the curative effect in rats with phosgene induced acute lung injury.
作者
金朝远
戴清霞
周芳庆
申捷
张琳
Jin Chaoyuan;Dai Qingxia;Zhou Qingfang;Shen Jie;Zhang Lin(Emergency&Intensive Care Centre for Chemical Accident of Jinshan Hospital Affiliated to Fudan University,Shanghai 201508,China)
出处
《中华劳动卫生职业病杂志》
CAS
CSCD
北大核心
2021年第2期110-113,共4页
Chinese Journal of Industrial Hygiene and Occupational Diseases
基金
国家自然科学基金(81801943)
上海市科学技术委员会项目(18411970200)。
关键词
热休克蛋白
间充质干细胞
光气
急性肺损伤
Heat shock protein
Mesenchymal stem cell
Phosgene
Acute lung injury