摘要
目的观察艾司氯胺酮对脂多糖诱导的心肌细胞内质网应激(ERS)与细胞凋亡的影响。方法2022年1—12月,采用脂多糖处理的H9c2细胞为心肌细胞毒模型,将细胞分为空白组(正常培养,不进行任何处理)、模型组(1 mg/L脂多糖处理H9c2细胞)及艾司氯胺酮组(8 mg/L艾司氯胺酮和1 mg/L脂多糖共同处理H9c2细胞)。处理24 h后,检测并比较各组H9c2细胞凋亡数量,细胞内凋亡蛋白胱天蛋白酶3(caspase-3)、胱天蛋白酶12(caspase-12)及多腺苷二磷酸核糖聚合酶1(PARP1),细胞内活性氧、丙二醛、超氧化物歧化酶(SOD)活性,培养上清中肿瘤坏死因子α(TNF-α)、白细胞介素(IL)-6和IL-1β蛋白水平及细胞内TNF-α、IL-6、IL-1βmRNA水平。检测并比较各组细胞ERS相关分子钙连蛋白、葡萄糖调节蛋白78(GRP78)、C/EBP同源蛋白(CHOP)、磷酸化蛋白激酶R样内质网激酶(p-PERK)、磷酸化真核翻译起始因子2α(p-eIF2α)蛋白水平。结果空白组、模型组及艾司氯胺酮组细胞晚期凋亡率分别为(1.88±0.17)%、(14.03±1.92)%、(5.68±1.43)%,钙连蛋白水平(荧光强度)分别为25.10±5.72、582.71±43.03、107.11±19.89。与空白组相比,模型组细胞凋亡细胞数量,活化PARP1/PARP1、活化caspase-3/caspase-3及活化caspase-12/caspase-12,细胞内活性氧、丙二醛及TNF-α、IL-6、IL-1β的mRNA水平,细胞培养上清TNF-α、IL-6、IL-1β蛋白水平,细胞内钙连蛋白、GRP78、CHOP蛋白水平,p-PERK、p-eIF2α蛋白水平显著增加(P<0.05),而艾司氯胺酮组上述指标水平显著低于模型组(P<0.05)。同时模型组SOD活性显著低于空白组(P<0.05),而艾司氯胺酮组SOD活性显著高于模型组(P<0.05),但低于空白组(P<0.05)。结论在H9c2细胞中,艾司氯胺酮能够抑制细胞凋亡,改善脂多糖诱导的ERS及炎症反应,这可能对减轻心肌细胞损伤有所帮助。
Objective To investigate the effects of esketamine on endoplasmic reticulum stress(ERS)and apoptosis induced by lipopolysaccharide(LPS)in cardiomyocytes.Methods From January to December 2022,an LPS-treated H9c2 cell model of cardiomyocyte injury was used.Cells were divided into three groups:a control group(normal culture without intervention),a model group(treated with 1 mg/L LPS),and an esketamine group(co-treated with 8 mg/L esketamine and 1 mg/L LPS).After 24 hours of treatment,the following parameters were measured and compared among groups:apoptotic cell count;intracellular levels of apoptotic proteins and their activated forms,including caspase-3,caspase-12,and poly(ADP-ribose)polymerase 1(PARP1);intracellular levels of reactive oxygen species(ROS),malondialdehyde(MDA),and superoxide dismutase(SOD)activity;protein levels of tumor necrosis factor-α(TNF-α),interleukin-6(IL-6),and interleukin-1β(IL-1β)in the culture supernatant;and intracellular mRNA levels of TNF-α,IL-6,and IL-1β.Additionally,protein levels of ERS-related molecules-calreticulin,glucose-regulated protein 78(GRP78),C/EBP homologous protein(CHOP),phosphorylated protein kinase R-like endoplasmic reticulum kinase(PERK),and phosphorylated eukaryotic initiation factor 2α(eIF2α)-were assessed.Results The late apoptosis rates in the control,model,and esketamine groups were(1.88±0.17)%,(14.03±1.92)%,and(5.68±1.43)%,respectively.Calreticulin levels(fluorescence intensity)were 25.10±5.72,582.71±43.03,and 107.11±19.89,respectively.Compared with the control group,the model group showed significant increases in apoptotic cell count;the ratios of activated to total PARP1,caspase-3,and caspase-12;intracellular levels of ROS and MDA;mRNA levels of TNF-α,IL-6,and IL-1β;supernatant protein levels of TNF-α,IL-6,and IL-1β;and intracellular protein levels of calreticulin,GRP78,CHOP,p-PERK,and peIF2α(all P<0.05).In the esketamine group,these indicators were significantly lower than those in the model group(P<0.05).Meanwhile,SOD activity in the model group was significantly lower than that in the control group(P<0.05),whereas it was significantly higher in the esketamine group than in the model group(P<0.05),but lower than that in the control group(P<0.05).Conclusion Esketamine inhibits apoptosis,ameliorates LPS-induced ERS,and suppresses inflammatory responses in H9c2 cells,suggesting a potential protective effect against cardiomyocyte injury.
作者
崔栋然
范方雪
武琼
王瑞
侯俊德
王志刚
CUI Dongran;FAN Fangxue;WU Qiong;WANG Rui;HOU Junde;WANG Zhigang(Department of Anesthesiology,HanDan Central Hospital,Handan,Hebei 056000,China;Department of Anesthesiology,HanDan First Hospital,Handan,Hebei 056000,China)
出处
《安徽医药》
2026年第3期486-491,共6页
Anhui Medical and Pharmaceutical Journal
基金
河北省医学科学研究课题计划(20210040)
邯郸市科学技术研究与发展计划项目(22422083044ZC)。
关键词
艾司氯胺酮
肌细胞
心脏
细胞损伤
凋亡
内质网应激
脂多糖
Esketamine
Myocytes,cardiac
Cell injury
Apoptosis
Endoplasmic reticulum stress
Lipopolysaccharide
