艾司氯胺酮通过上调miR-138-5p表达抑制LPS诱导的星形胶质细胞凋亡和炎症反应的分子机制研究  

Study on the molecular mechanism of esketamine inhibiting LPS-induced astrocyte apoptosis and inflammatory response by up-regulating the expression of miR-138-5p

作　　者：林玉美 符明君 陈基胜 LIN Yumei;FU Mingjun;CHEN Jisheng(Department of Anesthesiology,Haikou Third People's Hospital,Haikou,Hainan 571100,China)

机构地区：[1]海口市第三人民医院麻醉科,海南海口571100

出　　处：《国际检验医学杂志》2023年第8期908-913,共6页International Journal of Laboratory Medicine

基　　金：海南省医药卫生科研项目(1501032021A2001)。

摘　　要：目的探讨艾司氯胺酮对脂多糖(LPS)诱导的星形胶质细胞凋亡和炎症反应的影响及分子机制。方法星形胶质细胞HA1800分为对照(NC)组,0.5、1.0、2.0μg/mL LPS组,低、中、高剂量组,miR-138-5p组,miR-con组,高剂量+anti-miR-138-5p组、高剂量+anti-miR-con组;流式细胞术检测HA1800细胞凋亡情况;蛋白质印迹法检测裂解的半胱氨酸天冬氨酸蛋白酶3(Cleaved-caspase-3)蛋白表达;酶联免疫吸附试验(ELISA)检测肿瘤坏死因子-α(TNF-α)、白细胞介素(IL)-1β、IL-6水平;反转录-实时荧光定量PCR(RT-qPCR)检测微小RNA(miR)-138-5p表达水平。结果浓度越高的LPS诱导的星形胶质细胞HA1800凋亡率、Cleaved-caspase-3表达水平、TNF-α、IL-1β和IL-6水平升高(P<0.05)。剂量越高的艾司氯胺酮处理后,LPS诱导的HA1800细胞的凋亡率、Cleaved-caspase-3表达水平、TNF-α、IL-1β和IL-6水平降低,miR-138-5p表达水平升高(P<0.05)。过表达miR-138-5p后,LPS诱导的HA1800细胞的凋亡率、Cleaved-caspase-3表达水平、TNF-α、IL-1β和IL-6水平降低(P<0.05)。抑制miR-138-5p表达可以逆转艾司氯胺酮对LPS诱导的HA1800细胞凋亡和炎症反应的影响。结论艾司氯胺酮通过上调miR-138-5p表达抑制LPS诱导的星形胶质细胞凋亡和炎症反应。Objective To explore the effect of esketamine on lipopolysaccharide(LPS)-induced astrocyte apoptosis and inflammation and its molecular mechanism.Methods Astrocyte HA1800 was divided into NC group,0.5,1.0,2.0μg/mL LPS group,low,medium,and high dose group,miR-138-5p group,miR-con group,high-dose+anti-miR-138-5p group,high-dose+anti-miR-con group.Flow cytometry was used to detect HA1800 cell apoptosis,Western blot was used to detect the protein expression of Cleavage of cysteine aspartate protease 3(Cleaved-caspase-3),enzyme-linked immunosorbent assay(ELISA)was used to detect tumor necrosis factor-α(TNF-α),interleukin(IL)-1β,and IL-6 levels,and miR-138-5p expression level was detected by reverse transcription real-time fluorescent quantitative PCR(RT-qPCR).Results After induced with increased concentrations of LPS,the apoptosis rate,Cleaved-caspase-3 expression level,and the levels of TNF-α,IL-1βand IL-6 of astrocytes HA1800 were increased(P<0.05).After treatment with increased doses of esketamine,the apoptosis rate,Cleaved-caspase-3 expression level,and the levels of TNF-α,IL-1βand IL-6 of HA1800 cells induced by LPS were decreased,and the expression levels of miR-138-5p was increased(P<0.05).After overexpression of miR-138-5p,the apoptosis rate,Cleaved-caspase-3 expression level,and the levels of TNF-α,IL-1βand IL-6 of HA1800 cells induced by LPS were decreased(P<0.05).Inhibition of miR-138-5p expression could reverse the effect of esketamine on LPS-induced apoptosis and inflammation of HA1800 cells.Conclusion Esketamine inhibits LPS-induced astrocyte apoptosis and inflammation by up-regulating the expression of miR-138-5p.

关 键 词：艾司氯胺酮 微小RNA-138-5p 星形胶质细胞 凋亡 炎症反应 

分 类 号：R36[医药卫生—病理学]