Liu Ruizhen, Zhong Xinming, Zeng Jing, Huang Zhihua, Li Xiao, Xiao Hai, Chen Qin, Li Dongliang
Gannan Medical College, Ganzhou, Jiangxi 341000, P.R. China.
Key Laboratory of Cerebrovascular Diseases of Jiangxi Province, Ganzhou, Jiangxi 341000, P.R. China.
Int J Mol Med. 2017 Apr;39(4):1021-1028. doi: 10.3892/ijmm.2017.2915. Epub 2017 Mar 10.
This study aimed to observe the effects of 3'-daidzein sulfonate sodium (DSS) on ischemia-reperfusion-induced brain injury and to analyze the mechanisms responsible for neuronal apoptosis. Focal ischemias were induced in male Sprague-Dawley rats using middle cerebral artery occlusion. The rats were divided into 5 groups based on sham surgery or real occlusion, and treatment with different doses of DSS (0.5, 1.0 and 2.0 mg/kg) or normal saline (model group), injected preoperatively into the rats with cerebral occlusion. After 2 h of ischemia and 24 h of reperfusion, neurological deficit scores were evaluated using the Longa grade point standard. The infarct volume was measured using a triphenyl tetrazolium chloride staining technique. Blood-brain barrier (BBB) permeability was measured using the Evans blue (EB) content of brain tissues, while electron microscopy was used to observe ultrastructural changes. The expression levels of Bcl-2, Bax and caspase-3 were detected by an immunohistochemical method and western blot analysis. The neurological deficit in rats pre-treated with DSS at all doses decreased significantly (P<0.05) in comparison with the model group, as did the cerebral infarct volume ratios. The brain EB content was significantly reduced by the injection of DSS. The ultrastructural integrity of the rat BBB was significantly preserved in the DSS-treated groups in comparison with the model group. This was concomitant with the reduced swelling of astrocytes and pericytes in the BBB. The immunohistochemistry results revealed that DSS significantly enhanced the expression of Bcl-2, and inhibited the expression of Bax and caspase-3 in the brain in comparison to the model group. The number of apoptotic cells in the groups treated with DSS was reduced in comparison with similar areas in the model group. These findings suggest that DSS within a dosage range of 0.5-2.0 mg/kg provides significant protection from injury to the BBB induced by cerebral ischemia-reperfusion, as it exerts a neuroprotective effect by inhibiting apoptosis.
本研究旨在观察3'-大豆苷元磺酸钠（DSS）对缺血再灌注诱导的脑损伤的影响，并分析神经元凋亡的相关机制。采用大脑中动脉闭塞法在雄性Sprague-Dawley大鼠中诱导局灶性缺血。根据假手术或实际闭塞情况将大鼠分为5组，术前向脑闭塞大鼠注射不同剂量的DSS（0.5、1.0和2.0mg/kg）或生理盐水（模型组）。缺血2小时和再灌注24小时后，使用Longa评分标准评估神经功能缺损评分。采用氯化三苯基四氮唑染色技术测量梗死体积。使用脑组织的伊文思蓝（EB）含量测量血脑屏障（BBB）通透性，同时使用电子显微镜观察超微结构变化。通过免疫组织化学方法和蛋白质印迹分析检测Bcl-2、Bax和caspase-3的表达水平。与模型组相比，所有剂量的DSS预处理大鼠的神经功能缺损均显著降低（P<0.05），脑梗死体积比也显著降低。注射DSS后，脑EB含量显著降低。与模型组相比，DSS治疗组大鼠BBB的超微结构完整性得到显著保留。这与BBB中星形胶质细胞和周细胞肿胀的减轻相伴。免疫组织化学结果显示，与模型组相比，DSS显著增强了脑中Bcl-2的表达，并抑制了Bax和caspase-3的表达。与模型组的相似区域相比，DSS治疗组的凋亡细胞数量减少。这些发现表明，0.5-2.0mg/kg剂量范围内的DSS可显著保护脑缺血再灌注诱导的BBB损伤，因为它通过抑制凋亡发挥神经保护作用。
