摘要
目的研究右美托咪定(DEX)对脂多糖(LPS)诱导的大鼠肺泡Ⅱ型上皮(AT-Ⅱ)细胞核转录因子-κB(NF-κB)及水通道蛋白5(AQP-5)的影响。方法原代培养大鼠AT-Ⅱ细胞,建立LPS诱导急性肺损伤细胞(ALI)模型,实验分为对照组、LPS组(1μg·mL^-1)、LPS+DEX(0.1 ng·mL^-1、1 ng·mL^-1、10 ng·mL^-1)组。对照组置于正常条件下培养,LPS+DEX组在加入LPS前1 h分别加入相应浓度的右美托咪定,培养24 h后,MTT检测AT-Ⅱ细胞存活率,采用试剂盒测定各细胞上清液中乳酸脱氢酶(LDH)的含量,Real-time qPCR、Western blotting分别检测各组NF-κB及AQP-5的mRNA和蛋白表达水平。结果LPS可导致AT-Ⅱ细胞损伤,降低细胞存活率,增加LDH释放;右美托咪定可在0.1~10 ng·mL^-1内以浓度依赖性提高损伤细胞的存活率,减少LDH的释放,抑制LPS诱导的AT-Ⅱ细胞中NF-κB的活化,上调AQP-5的表达。结论右美托咪定对LPS所致AT-Ⅱ细胞损伤具有保护作用,其作用机制可能是下调NF-κB表达以及上调AQP-5表达。
Objective To investigate the effects of dexmedetomidine(DEX)pretreatment on the expressions of nuclear factor-κB(NF-κB)and aquaporin-5(AQP-5)in alveolar typeⅡ(AT-Ⅱ)cells induced by lipopolysaccharide(LPS)in rats.Methods The primary AT-Ⅱcells were isolated from rats previously injured with LPS as acute lung injury(ALI)models.The experiment was designed with five groups:the control group,the LPS group(1μg·mL^-1),and LPS+DEX(0.1 ng·mL^-1,1 ng·mL^-1,10 ng·mL^-1)groups.The control group was cultured under normal conditions,while the LPS+DEX groups were added with the corresponding concentration of dexmedetomidine 1 h before the addition of LPS.After 24 hours of culture,the survival rate of AT-Ⅱcells was detected by MTT colorimetry,and the level of lactate dehydrogenase(LDH)in the supernatant was measured by LDH kit.Real-time qPCR and Western blotting were used to detect the mRNA and protein expression of NF-κB and AQP-5 in each group.Results LPS induced injury in AT-Ⅱcells,decreased cell viability,and increased LDH release.Dexmedetomidine significantly increased the cell survival rate,decreased LDH release,inhibited NF-κB expression,and upregulated AQP-5 expression in a dose-dependent manner.Conclusion Dexmedetomidine had protective effects on LPS-induced AT-Ⅱcell injury.It may be related with the inhibition of NF-κB expression and up-regulation of AQP-5 expression.
作者
曾悦翔
文益云
杨金凤
孙辉平
ZENG Yuexiang;WEN Yiyun;YANG Jinfeng;SUN Huiping(Department of Anesthesiology,Hunan Cancer Hospital/the Affiliated Cancer Hospital of Xiangya School of Medicine,CentralSouth University,Changsha,Hunan,410013,China)
出处
《肿瘤药学》
CAS
2020年第2期191-195,213,共6页
Anti-Tumor Pharmacy
基金
湖南省自然科学基金(2019JJ40180)。
