摘要
目的探讨在脓毒症急性肺损伤(SA-ALI)中Sirt3对心磷脂合成酶1(CLS1)及心磷脂(CL)含量的影响。方法A549细胞随机分为对照组:细胞无处理;载体组:细胞转染空质粒;模型组:细胞经LPS(5μg/mL)刺激6 h建立细胞模型;Sirt3过表达组:细胞转染Sirt3质粒并LPS刺激建模。荧光素-光素酶法检测线粒体ATP、CL试剂盒检测CL、CCK-8检测细胞活力、Western blot检测CLS1表达。C57BL/6小鼠经气管内滴注LPS(5 mg/kg)建立SA-ALI模型,并随机分为对照组:小鼠气管内滴注生理盐水建立假模型;溶剂组:小鼠建立假模型后接受溶剂处理;模型组:小鼠建立SA-ALI模型接受溶剂处理;Sirt3抑制剂组:小鼠建立SA-ALI模型后接受Sirt3抑制剂处理。肺组织HE染色并行肺损伤评分;测定肺组织湿/干比重;Western blot检测肺组织CLS1表达。结果与对照组比较,模型组CLS1表达、CL含量、细胞内ATP含量及细胞活力显著下降,差异有统计学意义(P<0.05);与模型组比较,Sirt3过表达组细胞内CLS1、CL含量、ATP含量及细胞活力显著增加,差异有统计学意义(P<0.05)。与对照组比较,模型组小鼠肺组织内CLS1表达明显降低,肺损伤评分及湿/干比重显著增加,差异有统计学意义(P<0.05);与模型组比较,Sirt3抑制剂组小鼠肺组织内CLS1表达进一步下降,肺损伤评分及湿/干比重进一步增加,差异均有统计学意义(P<0.05)。结论Sirt3可能通过上调CLS1表达及CL合成从而减轻SA-ALI;针对CL合成的调控可能成为SAALI的潜在治疗靶点。
Objective To explore the effect of Sirt3 on cardiolipin synthase-1(CLS1)in the sepsis-associated acute lung injury(SA-ALI).Methods A549 cells were randomly divided into four groups:cells in control group received notreatment;cells in vector group were transfected with empty plasmid;cells in model group were stimulated with LPS(5μg/m L)for 6 h;cells in overexpression group were transfected with Sirt3 plasmid and received LPS stimulation.The ATP content was detected by a fluorescein-luciferase kit.The cardiolipin(CL)content was detected by a commercial kit.The cell viability was detected by cell counting kit-8(CCK-8)method.The CLS1 expression was detected by western blot.C57 BL/6 mice were intratracheal instilled with LPS(5 mg/kg)to establish the model of SA-ALI and randomly divided into four groups:mice in control groups were intratracheal instilled with saline to establish a sham-model.Mice in vehicle group were received a sham-model and treated with solvent;Mice in model group received SA-ALI model and treated with solvent;Mice in inhibitor group received SA-ALI model and treated with Sirt3 inhibitor.The pathological changes and lung injury score in lung were detected by HE staining.The lung edema was detected by the ratio of wet/dry of lung tissue.The expression of CLS1 was detected by western blot.Results In vitro,compared with the control group,the CLS1 expression,CL content,ATP content and cell viability of the model group were significantly decreased(P<0.05);compared with the model group,the the CLS1 expression,CL content,ATP content and cell viability of the Sirt3 overexpression group were significantly increased(P<0.05).In vivo,compared with the control group,the expression of CLS1 in the lung tissue of themodel group was significantly reduced and the lung injury score and wet/dry weight ratio was significantly increased(P<0.05);compared with the model group,the expression of CLS1 in the lung tissue of the inhibitor group was significantly reduced and the lung injury score and wet/dry weight ratio was si
作者
李志旺
蔡淑敏
李涛
陈仲清
LI Zhi-wang;CAI Shu-min;LI Tao;CHEN Zhong-qing(Department of Critical Care Medicine,Nanfang Hospital,Southern Medical University,Guangzhou,Guangdong 510515;Department of Critical Care Medicine,the First People's Hospital of Chenzhou,Chenzhou,Hunan 423000,China)
出处
《热带医学杂志》
CAS
2021年第1期1-5,F0004,共6页
Journal of Tropical Medicine
基金
国家自然科学基金(81500066)
湖南省自然科学基金(2018JJ3015)
南方医院院长基金(2017B004)。
