Jin C Y, Dai Q X, Zhou F Q, Shen J, Zhang L
Emergency & Intensive Care Centre for Chemical Accident of Jinshan Hospital Affiliated to Fudan University, Shanghai 201508, China.
Zhonghua Lao Dong Wei Sheng Zhi Ye Bing Za Zhi. 2021 Feb 20;39(2):110-113. doi: 10.3760/cma.j.cn.121094-20200104-00025.
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. 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 test. One way ANOVA was used for comparison between groups. The proliferation ability of MSCs transfected with HSP60[= (0.69±0.05) ] was significantly higher than that of MSCs not transfected with HSP60[= (0.27±0.02) ] (<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-α were significantly decreased, and the total protein content and total cell count in bronchoalveolar lavage fluid were less (<0.05) . 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.
探讨热休克蛋白60(HSP60)过表达对骨髓间充质干细胞(MSCs)能力的影响及其对光气诱导的急性肺损伤大鼠的治疗作用。通过腺病毒将HSP60转染到MSCs中。采用蛋白质免疫印迹法检测转染前后HSP60的表达。采用CCK-8法检测MSCs的活性,流式细胞术检测MSCs的凋亡能力,Transwell法观察MSCs的迁移能力。将60只SPF级雄性SD大鼠随机分为对照组、光气暴露组(吸入光气5分钟)、MSCs组(光气暴露,MSCs治疗组)和转染MSCs组(光气暴露,HSP60过表达MSCs治疗组)。通过肺病理切片观察肺的病理变化,检测肺湿干比、肺水肿程度、肺泡灌洗液的总细胞数和总蛋白含量,观察支气管肺泡灌洗液(BALF)和血清中肿瘤坏死因子-α(TNF-α)和白细胞介素-6(IL-6)的炎症变化。数据采用Graphpad Prism 8.0软件进行分析。配对比较采用非配对检验。组间比较采用单因素方差分析。转染HSP60的MSCs增殖能力[=(0.69±0.05)]显著高于未转染HSP60的MSCs[=(0.27±0.02)](<0.05)。与光气暴露组相比,MSCs组和转染MSCs组的肺水肿和炎症因子浸润减少。然而,与MSCs组相比,转染MSCs组的肺水肿程度明显改善,炎症因子IL-6和TNF-α水平明显降低,支气管肺泡灌洗液中的总蛋白含量和总细胞数较少(<0.05)。转染HSP60的MSCs可增强光气诱导的急性肺损伤大鼠的增殖、抗凋亡、迁移能力及治疗效果。