王明明 王磊 窦芳 李韦韦 文爱东 王婧雯

摘 要 目的：研究β-乳香酸对大鼠海马神经元细胞氧糖剥夺损伤的改善作用。方法：将大鼠海马神经元细胞分为正常对照组、模型组和β-乳香酸低、中、高浓度组（1、10、100 μmol/L），除正常對照组外，其余各组细胞加入相应含药培养基，并进行氧糖剥夺培养以复制氧糖剥夺损伤模型。采用MTT法检测细胞的存活率，采用化学比色法检测细胞上清液中乳糖脱氢酶（LDH）活性，采用Hoechst-PI染色法观察细胞形态的变化，采用流式细胞仪检测细胞早期凋亡率，采用Western blot法检测细胞中凋亡相关蛋白[B细胞淋巴瘤2（Bcl-2）、Bcl-2相关X蛋白（Bax）、激活型胱天蛋白酶3（cleaved caspase-3）]的表达。结果：与模型组比较，β-乳香酸中、高浓度组细胞存活率、Bcl-2蛋白表达水平均显著升高（P<0.01），细胞上清液中LDH活性、细胞早期凋亡率和细胞中cleaved caspase-3、Bax蛋白表达水平均显著降低 （P<0.05或P<0.01）;细胞核致密浓染、碎片状现象明显减少。结论：β-乳香酸可改善大鼠海马神经元细胞氧糖剥夺损伤，其作用机制可能与下调cleaved caspase-3、Bax蛋白表达，上调Bcl-2蛋白表达有关。

关键词 β-乳香酸;海马神经元细胞;氧糖剥夺;细胞凋亡

ABSTRACT   OBJECTIVE： To study the improvement effects of β-boswellic acid on hippocampal neurons cells injury of rats induced by oxygen-glucose deprivation. METHODS： The hippocampal neurons cell of rats were divided into normal control group， model group and β-boswellic acid low-concentration， medium-concentration and high-concentration groups （1， 10， 100 μmol/L）. Except for normal control group， other groups were cultured with relevant medium and given oxygen glucose deprivation to induce oxygen-glucose deprivation induced injury model. MTT assay was adopted to detect cell viability. Chemical colorimetry was used to detect LDH activity in cell culture supernatant. Hoechst-PI staining was used to detect the morphology change of cells. Flow cytometry was used to detect early apoptosis rate of cells. The expression of apoptosis-related protein （Bcl-2， Bax and cleaved caspase-3） were detected by Western blot. RESULTS： Compared with model group， the survival rate of cells and protein expression of Bcl-2 were increased significantly in β-boswellic acid medium-concentration and high-concentration groups （P<0.01）， while LDH activity， early apoptosis rate， protein expression of cleaved caspase-3 and Bax were all decreased significantly （P<0.05 or P<0.01）. The densely stained nuclei and fragmentation decreased significantly. CONCLUSIONS： β-boswellic acid can relieve oxygen-glucose deprivation induced injury of hippocampal neurons cells， the mechanism of which may be associated with down-regulating the protein expression of cleaved caspase-3 and Bax and up-regulating the protein expression of Bcl-2.

KEYWORDS   β-boswellic acid; Hippocampal neurons cells; Oxygen-glucose deprivation; Cell apoptosis

脑卒中是引起人类死亡和残疾的第二大病因，具有高发病率、高病死率、高致残率、高复发率及经济负担重的特点，严重影响国民健康;其中，80%的患者为缺血性卒中[1]。缺血性脑卒中是因大脑动脉血流短暂或持久性的闭塞导致脑供血不足或中断，进而诱发一系列级联反应，造成缺血区脑组织不可逆的损伤，导致神经功能缺陷[2]。海马组织是中枢神经系统的重要组成部分，是脑组织中对缺血缺氧最敏感的部位之一[3-4]，因此，抑制海马神经元细胞凋亡可有效减轻缺血性脑损伤的程度，进而防治缺血性脑卒中[5]。

乳香为橄榄科植物乳香树 Boswellia carterii Birdw.及同属B. bhaur dajiana Birdw.树皮渗出的树脂，目前已被欧洲药品管理局列为治疗脑癌水肿的潜在治疗药物[6]，其中β-乳香酸是其重要的药效物质[7]。……