Biphalin激活PI3K/Akt通路减轻氧糖剥夺/复氧诱导的神经元损伤  

Biphalin alleviated oxygen-glucose deprivation/reoxygenation-induced neuronal injury by activating PI3K/Akt signaling pathway

作　　者：陈玲 张锦佳 黄宇钧 陆媛 闵加威 CHEN Ling;ZHANG Jinjia;HUANG Yujun;LU Yuan;MIN Jiawei(College of Biomedical Engineering,South-Central Minzu University,Wuhan 430074,China)

机构地区：[1]中南民族大学生物医学工程学院,武汉430074

出　　处：《中南民族大学学报(自然科学版)》2025年第3期357-364,共8页Journal of South-Central Minzu University(Natural Science Edition)

基　　金：国家自然科学基金资助项目(82301574);湖北省自然科学基金资助项目(2024AFB875);中央高校基本科研业务费专项资金资助项目(CZQ23030);中南民族大学科学研究基金资助项目(YZZ19018)。

摘　　要：为了探究biphalin对氧糖剥夺/复氧(oxygen-glucose deprivation/reoxygenation,OGD/R)诱导的原代皮层神经元细胞损伤的影响及其机制,将神经元细胞分5组处理:对照组常规培养;OGD/R组仅构建OGD/R损伤模型;OGD/R+biphalin组细胞于造模过程中加入不同浓度biphalin(0.01、0.1、1、10 nmol/L);OGD/R+biphalin+纳洛酮组细胞在造模过程中用0.1 nmol/L biphalin和0.1 mmol/L纳洛酮处理;OGD/R+biphalin+LY294002组细胞在造模过程中用0.1 nmol/L biphalin和20μmol/L LY294002处理.采用CCK-8法、乳酸脱氢酶(LDH)释放法、碘化丙啶(PI)染色法测定biphalin对神经元细胞的影响.采用蛋白质印迹法检测Bcl-2、Bax、cleaved-caspase-3、p-PI3K、PI3K、p-Akt和Akt的蛋白表达情况.经CCK-8法检测细胞活力,选定0.1 nmol/L biphalin为最佳给药剂量.与对照组相比,模型组细胞活力明显下降,细胞凋亡率和LDH释放量增加,cleaved-caspase-3和Bax表达量升高.Biphalin处理组细胞较模型组存活率显著提高,LDH释放量减少,cleaved-caspase-3和Bax表达量减少,Bcl-2水平和p-PI3K/PI3K、p-Akt/Akt比值升高.而阿片拮抗剂纳洛酮和PI3K抑制剂LY294002可以显著逆转biphalin的上述作用效果.这表明biphalin通过阿片受体激活PI3K/Akt通路从而减轻了OGD/R介导的神经元损伤.To investigate the effects and mechanism of biphalin on primary cortical neuronal injury induced by oxygenglucose deprivation/reoxygenation(OGD/R),neurons were randomly divided into five groups,namely the control group(without intervention),the OGD/R group(underwent OGD/R only),the OGD/R+biphalin group(treated with 0.01,0.1,1,and 10 nmol/L biphalin during OGD/R),the OGD/R+biphalin+naloxone group(treated with 0.1 nmol/L biphalin and 0.1 mmol/L naloxone during OGD/R),the OGD/R+biphalin+LY294002 group(treated with 0.1 nmol/L biphalin and 20μmol/L LY294002 during OGD/R).Cell counting kit-8(CCK-8),lactate dehydrogenase(LDH)assays and propidium iodide(PI)staining were performed to analyse the effects of biphalin on primary cortical neurons.The protein expression levels of Bcl-2,Bax,cleaved-caspase-3,p-PI3K,PI3K,p-Akt and Akt were measured by Western blotting.Due to the greatest neuroprotection of biphalin was observed at 0.1 nmol/L,this concentration was chosen for subsequent studies.The decreased cell viability and increased cell death and LDH release were observed after OGD/R compared to the control group.The levels of cleaved-caspase-3 and Bax significantly increased in OGD/R group compared with the control group.Compared with the OGD/R group,biphalin significantly increased the survival rate and the level of Bcl-2 and the ratios of p-PI3K/PI3K and p-Akt/Akt,reduced LDH release and the protein levels of cleaved-caspase-3 and Bax.However,both of them were mostly suppressed by the opioid antagonist naloxone and the PI3K inhibitor LY294002 respectively.These results suggest that biphalin protected against OGD/R injury in primary cortical neurons by activating PI3K/Akt signaling pathway via opioid receptor.

关 键 词：biphalin肽 新生儿缺氧缺血性脑损伤 氧糖剥夺/复氧(OGD/R) PI3K/AKT通路 原代皮层神经元细胞 

分 类 号：R965.1[医药卫生—药理学]