PDF(Pubmed)
Abstract：
Cellular stress associated with static-cold storage (SCS) and warm reperfusion of donor lungs can contribute to ischemia-reperfusion (IR) injury during transplantation. Adding cytoprotective agents to the preservation solution may be conducive to reducing graft deterioration and improving post-transplant outcomes.
SCS and warm reperfusion were simulated in human lung epithelial cells (BEAS-2B) by exposing cells to low potassium dextran glucose solution at 4 °C for different periods and then switching back to serum-containing culture medium at 37 °C. Transcriptomic analysis was used to explore potential cytoprotective agents. Based on its results, cell viability, caspase activity, cell morphology, mitochondrial function, and inflammatory gene expression were examined under simulated IR conditions with or without thyroid hormones (THs).
After 18 h SCS followed by 2 h warm reperfusion, genes related to inflammation and cell death were upregulated, and genes related to protein synthesis and metabolism were downregulated in BEAS-2B cells, which closely mirrored gene profiles found in thyroid glands of mice with congenital hypothyroidism. The addition of THs (T3 or T4) to the preservation solution increases cell viability, inhibits activation of caspase 3, 8 and 9, preserves cell morphology, enhances mitochondrial membrane potential, reduces mitochondrial superoxide production, and suppresses inflammatory gene expression.
Adding THs to lung preservation solutions may protect lung cells during SCS by promoting mitochondrial function, reducing apoptosis, and inhibiting pro-inflammatory pathways. Further in vivo testing is warranted to determine the potential clinical application of adding THs as therapeutics in lung preservation solutions.
SCS and warm reperfusion were simulated in human lung epithelial cells (BEAS-2B) by exposing cells to low potassium dextran glucose solution at 4 °C for different periods and then switching back to serum-containing culture medium at 37 °C. Transcriptomic analysis was used to explore potential cytoprotective agents. Based on its results, cell viability, caspase activity, cell morphology, mitochondrial function, and inflammatory gene expression were examined under simulated IR conditions with or without thyroid hormones (THs).
After 18 h SCS followed by 2 h warm reperfusion, genes related to inflammation and cell death were upregulated, and genes related to protein synthesis and metabolism were downregulated in BEAS-2B cells, which closely mirrored gene profiles found in thyroid glands of mice with congenital hypothyroidism. The addition of THs (T3 or T4) to the preservation solution increases cell viability, inhibits activation of caspase 3, 8 and 9, preserves cell morphology, enhances mitochondrial membrane potential, reduces mitochondrial superoxide production, and suppresses inflammatory gene expression.
Adding THs to lung preservation solutions may protect lung cells during SCS by promoting mitochondrial function, reducing apoptosis, and inhibiting pro-inflammatory pathways. Further in vivo testing is warranted to determine the potential clinical application of adding THs as therapeutics in lung preservation solutions.
摘要：
背景：与供肺的静冷储存（SCS）和热再灌注相关的细胞应激可导致移植过程中的缺血再灌注（IR）损伤。向保存溶液中添加细胞保护剂可能有助于减少移植物退化并改善移植后结果。
方法：在人肺上皮细胞（BEAS-2B）中模拟SCS和热再灌注，方法是将细胞暴露于4°C的低钾葡聚糖葡萄糖溶液中不同时间，然后在37°C下切换回含有血清的培养基。转录组学分析用于探索潜在的细胞保护剂。根据其结果,细胞活力，caspase活性，细胞形态学,线粒体功能，在有或没有甲状腺激素（THs）的模拟IR条件下检查炎症基因表达。
结果：经过18小时SCS，然后经过2小时的热再灌注，与炎症和细胞死亡相关的基因上调，BEAS-2B细胞中与蛋白质合成和代谢相关的基因下调，这与先天性甲状腺功能减退症小鼠甲状腺中发现的基因谱密切相关。添加TH（T3或T4）到保存溶液增加细胞活力，抑制caspase3，8和9的激活，保留细胞形态，增强线粒体膜电位，减少线粒体超氧化物的产生，并抑制炎症基因表达。
结论：在肺保存液中加入THs可能通过促进线粒体功能来保护SCS期间的肺细胞，减少细胞凋亡，并抑制促炎途径。需要进一步的体内测试以确定在肺保存溶液中添加THs作为治疗剂的潜在临床应用。
方法：在人肺上皮细胞（BEAS-2B）中模拟SCS和热再灌注，方法是将细胞暴露于4°C的低钾葡聚糖葡萄糖溶液中不同时间，然后在37°C下切换回含有血清的培养基。转录组学分析用于探索潜在的细胞保护剂。根据其结果,细胞活力，caspase活性，细胞形态学,线粒体功能，在有或没有甲状腺激素（THs）的模拟IR条件下检查炎症基因表达。
结果：经过18小时SCS，然后经过2小时的热再灌注，与炎症和细胞死亡相关的基因上调，BEAS-2B细胞中与蛋白质合成和代谢相关的基因下调，这与先天性甲状腺功能减退症小鼠甲状腺中发现的基因谱密切相关。添加TH（T3或T4）到保存溶液增加细胞活力，抑制caspase3，8和9的激活，保留细胞形态，增强线粒体膜电位，减少线粒体超氧化物的产生，并抑制炎症基因表达。
结论：在肺保存液中加入THs可能通过促进线粒体功能来保护SCS期间的肺细胞，减少细胞凋亡，并抑制促炎途径。需要进一步的体内测试以确定在肺保存溶液中添加THs作为治疗剂的潜在临床应用。
