医学分子生物学杂志 ›› 2024, Vol. 21 ›› Issue (6): 568-575.doi: 10.3870/j.issn.1672-8009.2024.06.011

• 论著 • 上一篇    下一篇

过表达 lincRNA-p21 的间充质干细胞对小鼠急性肺损伤的修复作用研究 #br#

  

  1. 新疆医科大学第二附属医院1重症医学一科,2重症医学二科,3感染科 乌鲁木齐市, 830054
  • 出版日期:2024-11-30 发布日期:2024-12-09
  • 基金资助:
    新疆神经系统疾病研究重点实验室 (No. XJDX1711-2250)

Reparative Effect of Mesenchymal Stem Cells Overexpressing LincRNA-p21 on Acute Lung Injury in Mice #br#

  1. 1First Department of Intensive Care Medicine,2Second Department of Intensive Care Medicine,3Department of Infection, Second Affiliated Hospital of Xinjiang Medical University, Urumqi, 830054, China
  • Online:2024-11-30 Published:2024-12-09

摘要: 目的 探讨基因间长链非编码 RNA ( long intergenic non-coding RNA, lincRNA) -p21 过表达的间充质干细胞 (mesenchymal stem cells, MSCs) 对小鼠急性肺损伤 (acute lung injury, ALI) 的修复作用方法 70 只雄性 C57BL / 6 小鼠随机分为 7 , 每组 10 , 分别为假手术组、 ALI 、 MSC + ALI 、 p21- MSC + ALI 空载体-MSC + ALI 、 p21 + ALI 空载体 + ALI 实时荧光定量 PCR ( qRT-PCR) 法检测各组肺组织中 lincRNA-p21 的表达。 ELISA 法检测肺泡灌洗液中 TNF-α、 IL-1β、 IL-8、 IL-6、 IL-10、 IL-13、 IL-1Ra、 caspase3 的表达检测动脉血氧分压 ( PaO2 )、 肺系数 ( lung index, LI)、 肺通透指数 ( lungpermeability index, LPI)、 氧合指数 (oxygenation index, OI) 的影响蛋白质印迹法检测肺组织中 Bcl-2、 Bax、caspase3、 cleaved-caspase3 的表达结果 与假手术组比较, ALI lincRNA-p21、 IL-10、 IL-13、 IL-1Ra、 Bcl-2、PaO2OI 的水平都下调, LI、 LPI、 TNF-α、 IL-1β、 IL-8、 IL-6、 Bax、 caspase3、 cleaved-caspase3 的水平都上调,凋亡细胞比例增加 (P<0. 05)。 ALI 组比较, MSC + ALI IL-10、 IL-13、 IL-1Ra、 Bcl-2、 PaO2OI 的水平都上调, LI、 TNF-α、 IL-1β、 IL-8、 IL-6、 Bax、 caspase3、 cleaved-caspase3 的水平都下调, 凋亡细胞比例减少(P<0. 05), lincRNA-p21 的表达和 LI 的水平变化不明显 (P >0. 05)。 与空载体-MSC + ALI 组比较, p21-MSC+ ALI 组中 lincRNA-p21、 IL-10、 IL-13、 IL-1Ra、 Bcl-2、 PaO2OI 的水平都上调, LI、 LPI、 TNF-α、 IL-1β、 IL-8、 IL-6、 Bax、 caspase3、 cleaved-caspase3 的水平都下调, 凋亡细胞比例减少 (P< 0. 05)。 与空载体 + ALI 组比较, p21 + ALI 组中 lincRNA-p21、 IL-10、 IL-13、 IL-1Ra、 Bcl-2、 PaO2OI 的水平都上调 (P< 0. 05), LI、 LPI、TNF-α、 IL-1β、 IL-8、 IL-6、 Bax、 caspase3、 cleaved-caspase3 的水平都下调, 凋亡细胞比例减少 (P< 0. 05)。 结论 过表达 lincRNA-p21 MSCs ALI 小鼠的炎症损伤和肺部细胞凋亡具有抑制作用

关键词:

基因间长链非编码 RNA-p21, 间充质干细胞, 急性肺损伤, 小鼠, 炎症, 凋亡

Abstract: Objective To investigate the reparative effect of mesenchymal stem cells (MSCs)overexpressing long intergenic non-coding RNA ( lincRNA) -p21 on acute lung injury ( ALI) inmice. Methods Seventy male C57BL / 6 mice were randomly divided into 7 groups, with n = 10 ineach group. The 7 groups were as follows: sham group, ALI group, MSC + ALI group, p21-MSC + ALI group, empty vector-MSC + ALI group, p21 + ALI group, and empty vector + ALI group. Real-time fluorescence quantitative PCR (qRT-PCR) was used to detect the expression level of lincRNA-p21 in lung tissues of each group. ELISA was used to detect the levels of TNF-α, IL- 1β, IL-8, IL-6, IL-10, IL-13, IL-1Ra, and caspase3 in bronchoalveolar lavage fluid. The effects on arterial blood oxygen pressure ( PaO2 ), lung index ( LI), lung permeability index (LPI), and oxygenation index (OI) were evaluated. Flow cytometry was used to detect and ana
lyze the proportion of apoptotic cells in lung tissues. Western blotting was used to detect the expression levels of Bcl-2, Bax, caspase3, and cleaved-caspase3 in lung tissues. Results The levels of lincRNA-p21, IL-10, IL-13, IL-1Ra, Bcl-2, and the values of PaO2 and OI in the ALI group were decreased, while LI, LPI, TNF-α, IL-1β, IL-8, IL-6, Bax, caspase3, cleaved-caspase3 were upregulated, and the proportion of apoptotic cells was increased when compared with those inthe sham group ( all P < 0. 05). The levels of IL-10, IL-13, IL-1Ra, Bcl-2, and the values ofPaO 2and OI were all increased in the MSC + ALI group, and the levels of TNF-α, IL-1β, IL-8, IL-6, Bax, caspase3 and cleaved caspase3, and the values of LI were all decreased when compared with those in the ALI group, the proportion of apoptotic cells was decreased (all P< 0. 05).The expression level of lincRNA-p21 and the value of LPI had no significant change between abovegroups (P > 0. 05). The levels of lincRNA-p21, IL-10, IL-13, IL-1Ra, Bcl-2, and the valuesof PaO 2and OI were increased, while values of LI, LPI, and levels of TNF-α, IL-1β, IL-8, IL- 6, Bax, caspase3, cleaved-caspase3 were decreased, and the proportion of apoptotic cells was decreased in the p21-MSC + ALI group when compared with those in the empty vector-MSC + ALIgroup (all P< 0. 05). The levels of lincRNA-p21, IL-10, IL-13, IL-1Ra, Bcl-2, and the values of PaO 2and OI in the p21 + ALI group were increased (all P < 0. 05), and the levels of TNF-α,IL-1β, IL-8, IL-6, Bax, caspase3, and cleaved caspase3, and the values of LI, LPI were decreased, and the proportion of apoptotic cells was decreased when compared with those in the emptyvector + ALI group (all P< 0. 05). Conclusion MSCs overexpressing lincRNA-p21 have an inhibitory effect on inflammatory injury and lung cell apoptosis in ALI mice.

Key words:

long non-coding RNA-p21, mesenchymal stem cells, acute lung injury, mice, inflammation, apoptosis

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