首页|CaMKⅡγ和CaMKⅡδ通过PI3K/Akt/Erk信号通路减轻小鼠神经元缺血再灌注损伤

CaMKⅡγ和CaMKⅡδ通过PI3K/Akt/Erk信号通路减轻小鼠神经元缺血再灌注损伤

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  • 国家科技期刊平台
  • NETL
  • NSTL
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目的 观察钙/钙调蛋白依赖性激酶Ⅱ(CaMKⅡ)的同工型CaMKⅡγ和CaMKⅡδ对鼠神经元细胞缺血缺氧再灌注损伤的影响，并探究其作用机制。方法 分离胚胎期第18天胎鼠大脑用于提取原代神经元，在5%CO2、37℃条件下培养，分为常规培养的对照组、空白对照组(si-NT)、CaMKⅡγ敲除组(si-CAMK2G)和CaMKⅡδ敲除组(si-CAMK2D)。研究组神经元转染处理后，更换为无糖培养基，并将其置于缺氧环境中模拟氧糖剥夺(OGD/R)条件，持续1h，随后复原标准培养环境。通过对神经元裂解物行免疫蛋白印迹检测磷脂酰肌醇-3-激酶/细胞外信号调节激酶(PI3K/Akt/Erk)信号通路构件的表达量，并建立小鼠大脑中动脉闭塞(MCAO)模型，通过对比假手术组(Sham)(n=25)和MCAO组(n=25)PI3K/Akt/Erk信号通路表达来进行验证。结果 si-CAMK2G组的胎鼠神经元细胞在OGD/R 12、24、48、72 h的生存率明显低于si-NT组(P<0。01或0。001)，并可逆转OGD/R介导的胎鼠神经元细胞CaMKⅡγ表达上调。si-CAMK2G组和si-CAMK2D组与si-NT组相比，PI3K/Akt/Erk信号通路表达受到明显抑制(P<0。01)。在MCAO模型中，MCAO组小鼠脑CaMKⅡδ和CaMKⅡγ的表达显著增加并激活了PI3K/Akt/Erk信号通路，CaMKⅡδ和CaMKⅡγ，Erk、磷酸化Erk、Akt和磷酸化Akt在MCAO造模成功再灌注后24、48、72和96h的表达显著高于Sham组再灌注24h(P<0。05，0。01或0。0001)。结论 CaMKⅡγ和CaMKⅡδ在神经元细胞发生缺血缺氧损伤时的神经保护作用可能是通过PI3K/Akt/Erk信号通路介导的。
PI3K/Akt/Erk signaling pathway mediates neuroprotection of CaMKⅡγ and CaMKⅡδ against ischemic reperfusion injury in mice
Objective To observe neuroprotective effects of Ca2+/calmodulin-dependent kinase Ⅱ(CaMKⅡ)γ and CaMkⅡ δ against acute neuronal ischemic reperfusion injury in mice and explore the underlying mechanism.Methods Primary cultures of brain neurons isolated from fetal mice(gestational age of 18 days)were transfected with two specific siRNAs(si-CAMK2G and si-CAMK2D)or a control sequence(si-NT).After the transfection,the cells were exposed to oxygen-glucose deprivation/reperfusion(OGD/R)conditions for 1 h followed by routine culture.The expressions of phosphatidylinositol-3-kinase/extracellular signal-regulated kinase(PI3K/Akt/Erk)signaling pathway components in the neurons were detected using immunoblotting.The expressions of the PI3K/Akt/Erk signaling pathway proteins were also detected in the brain tissues of mice receiving middle cerebral artery occlusion(MCAO)or sham operation.Results The neuronal cells transfected with si-CAMK2G showed significantly lower survival rates than those with si-NT transfection at 12,24,48,and 72 h after OGD/R(P<0.01),and si-CAMK2G transfection inhibited OGD/R-induced upregulation of CaMKⅡγ expression.Compared to si-NT,transfection with si-CAMK2G and si-CAMK2D both significantly inhibited the expressions of PI3K/Akt/Erk signaling pathway components(P<0.01).In the mouse models of MCAO,the expressions of CaMKⅡδ and CaMKⅡγ were significantly increased in the brain,where activation of the PI3K/Akt/Erk signaling pathway was detected.The expression levels of CaMKⅡδ,CaMKⅡγ,Erk,phosphorylated Erk,Akt,and phosphorylated Akt were all significantly higher in MCAO mice than in the sham-operated mice at 24,48,72,and 96 h after reperfusion(P<0.05).Conclusion The neuroprotective effects of CaMKⅡδ and CaMKⅡγ against acute neuronal ischemic reperfusion injury are mediated probably by the PI3K/Akt/Erk pathway.

cerebral ischemia/reperfusion injuryCa2+/calmodulin-dependent kinaseⅡneuroprotectionphosphatidylinositol-3-kinaseextracellular signal-regulated kinasesignaling pathway

刘昊铭、林子诗、叶靖

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南方医科大学南方医院麻醉科,广东 广州 510515

佛山市第一人民医院麻醉科,广东 佛山 528000

南方医科大学珠江医院麻醉科,广东 广州 510260

脑缺血再灌注损伤 钙/钙调蛋白依赖性激酶Ⅱ 神经保护 磷脂酰肌醇-3-激酶 细胞外信号调节激酶 信号通路

广东省自然科学基金广东省高等教育教学改革项目

2019A1515011190

2024

10.12122/j.issn.1673-4254.2024.03.18

南方医科大学学报
南方医科大学

南方医科大学学报

CSTPCD北大核心
影响因子：1.654
ISSN：1673-4254
年,卷(期)：2024.44(3)
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