目的 探讨右美托咪定(Dex)对皮质神经元氧糖剥夺/复氧(OGD/R)损伤的神经保护作用及其可能机制.方法 培养孕18 d的SD大鼠胎鼠皮质神经元细胞,随机分为对照组(Sham组,加入有糖细胞外液置于正常培养箱中培养)、模型组(OGD/R组,加入无糖细胞外液,置于37 ℃厌氧箱中培养制备OGD/R细胞模型)、阳性药物对照组(OGD/R+LW6组,OGD/R模型细胞加入1 mmol/L的LW6处理1 h)、OGD/R+低浓度Dex组(OGD/R模型细胞加入0.1 μmol/L Dex处理1 h)、OGD/R+高浓度Dex组(OGD/R模型细胞加入1.0 μmol/L Dex处理1 h).应用CCK-8比色法、免疫荧光方法检测各组细胞的存活率,Western blot方法检测各组低氧诱导因子1α(HIF-1α)、激活转录因子-6(ATF-6)及下游靶点C/EBP同源蛋白(CHOP)的表达水平.结果 CCK-8比色及免疫荧光染色结果显示,各组神经元存活率差异有显著性(F=63.46,P＜0.05);两两比较显示,O GD/R组神经元存活率较Sham组显著降低(P＜0.05),OGD/R+高浓度Dex组神经元存活率较OGD/R组显著提高(P＜0.05).Western blot结果显示,各组神经元HIF-1α、ATF-6、CHOP蛋白表达差异有显著性(F=80.30～155.36,P＜0.05);两两比较显示,OGD/R组HIF-1α、ATF-6、CHOP蛋白表达较Sham组升高(P＜0.05),OGD/R+高浓度Dex组HIF-1α、ATF-6、CHOP蛋白表达较OGD/R组降低(P＜0.05).结论 Dex通过抑制HIF-1α表达对OGD/R损伤的神经元模型发挥保护作用.
Objective To investigate the neuroprotective effects of dexmedetomidine(Dex)against oxygen-glucose depriva-tion/reoxygenation(OGD/R)injury of cortical neurons and its possible mechanism of action.Methods The cortical neurons of fetal Sprague-Dawley rats(gestational age,18 d)were cultured and then were randomly divided into sham group(cultured in a sugar-containing medium in normal incubator),OGD/R group(cultured in a sugar-free medium in an anaerobic incubator at 37 ℃to prepare a neuronal OGD/R injury model),OGD/R+LW6 group(OGD/R neurons treated with 1 mmol/L LW6 for 1 h as a po-sitive control),OGD/R+low-concentration Dex group(OGD/R neurons treated with 0.1 μmol/L Dex for 1 h),and OGD/R+high-concentration Dex group(OGD/R neurons treated with 1.0 μmol/L Dex for 1 h).The survival rate of cells was determined by co-lorimetric assay with Cell Counting Kit-8(CCK-8)and immunofluorescence assay.The expression levels of hypoxia-inducible factor-1α(HIF-1α),activating transcription factor 6(ATF-6),and the downstream target C/EBP homologous protein(CHOP)were mea-sured by Western blot.Results According to CCK-8 and immunofluorescence assay results,there was a significant diffe-rence in the survival rate of neurons between the groups(F=63.46,P＜0.05).Pairwise comparison showed that the survival rate in the OGD/R group was significantly lower than that in the sham group(P＜0.05);and the survival rate of the OGD/R+high-con-centration Dex group was significantly increased compared with that of the OGD/R group(P＜0.05).Western blot results showed significant differences in the expression of HIF-1α,ATF-6,and CHOP proteins(F=80.30-155.36,P＜0.05).Pairwise compari-son showed that the expression of HIF-1α,ATF-6,and CHOP was significantly higher in the OGD/R group than in the sham group(P＜0.05);and the OGD/R+high-concentration Dex group showed significantly lower expression of HIF-1α,ATF-6,and CHOP compared with the OGD/R group,the differences are statistically significant(P＜0.05).Conclusion Dex inhibits HIF-1α expression to exert protective effects in the neuronal model of OGD/R injury.
张雅瑞;侯庆明
青岛大学神经再生与康复研究院,山东青岛 266071
基础医学
右美托咪定;低氧诱导因子1α;氧糖剥夺/复氧损伤;激活转录因子6;神经保护
dexmedetomidine;hypoxia-inducible factor-1alpha;oxygen glucose deprivation/reoxygenation injury;activating transcription factor 6;neuroprotection
《青岛大学学报（医学版）》 2024 (001)
12-16 / 5
国家自然科学基金资助项目(8217051911)
10.11712/jms.2096-5532.2024.60.004
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