摘要
目的探讨灯盏花素(Bre)通过调控NLRP3炎症小体活化抑制慢性脑缺血(CCI)大鼠神经元细胞焦亡与凋亡作用。方法雄性SD大鼠随机分为假手术组、2VO结扎组、2VO结扎+灯盏花素(2.5、5、10 mg/kg)组。大鼠双侧颈总动脉永久结扎(2VO)构建CCI动物模型,Morris水迷宫检测大鼠学习记忆能力;苏木精-伊红染色(HE)和TUNEL法分别检测大鼠脑组织病理学变化与细胞凋亡,Western blot法检测大鼠海马组织中NLRP3、Caspase 1、白细胞介素(IL)-1β、IL-6、cleaved Caspase-3等蛋白表达。结果与假手术组相比,2VO结扎组大鼠认知功能下降、主要表现为潜伏期延长、目标象限停留时间缩短、穿越次数减少(P<0.01),皮层与海马神经元凋亡增加、变性与坏死加重,同时海马组织NLRP3炎症小体激活,Caspase 1、IL-6、IL-1β蛋白表达上调,Caspase-3蛋白激活,进一步促进神经元凋亡。与2VO结扎组相比,Bre各剂量组大鼠发现平台的潜伏期缩短,目标象限停留时间长、穿越次数增多;病理学检测发现CCI大鼠皮层与海马神经元损伤减轻,神经元数量增多,核固缩缓解,凋亡减少,细胞形态改善。另外灯盏花素可显著抑制海马组织NLRP3炎症小体活化,下调Caspase 1、IL-6和IL-1β蛋白表达,抑制Caspase-3蛋白的活化,抑制神经元凋亡。结论灯盏花素可显著改善CCI大鼠的认知功能,减轻缺血神经元的病理性损伤,其机制可能与抑制脑组织NLRP3炎症小体活化与细胞焦亡途径有关。
Objective To investigate the inhibitory effect of breviscapine(Bre)on pyroptosis and apoptosis of neu-ronal cells in chronic cerebral ischemia(CCI)rats by regulating the activation of NLRP3 inflammasome.Methods Male SD rats were randomly divided into sham group,2VO group,2VO+breviscapine(2.5,5,10 mg/kg)groups.The model of CCI was established by permanently ligating the bilateral common carotid arteries(2VO)in rats.The learning and memory ability of rats were detected by Morris water maze test.The pathological changes and apoptosis of brain tissue were detected by HE and Tunel staining respectively.NLRP3,Caspase 1,interleukin(IL)-1β,IL-6 and cleaved Caspase-3 proteins expression levels were detected by Western blot.Results Compared with the sham group,the spatial learning and memory ability was disordered,the latency was significantly pro-longed,the target quadrant dwell time and the number of crossing target quadrants significantly reduced in the 2VO group rats(P<0.01).The apoptosis,degeneration and necrosis have been increased in cortical and hippocampal neurons in the 2VO group rats.Also,the NLRP3 inflammasome in the hippocampus was activated,the expressions of Caspase 1,IL-6 and IL-1βprotein were up-regulated,and Caspase-3 protein was activated,which further pro-moted neuronal apoptosis in the 2VO group rats.Compared with the 2VO group,Bre significantly reduced the la-tency of the platform found,increased the target quadrant residence time and the number of crossing target quad-rants in rats.Bre reduced apoptosisand the pathological damage of cortical and hippocampal neurons,increased the number of neuronal cells,and improved neuronal cell morphology and alleviated nuclear pyknosis in CCI rats.In addition,Bre significantly inhibited the activation of NLRP3 inflammasome in hippocampus,down-regulated the ex-pression of Caspase 1,IL-6 and IL-1β,inhibited neuronal apoptosis and the activation of Caspase-3 proteinand.Conclusion Bre can significantly improve the cognitive function of CCI rats,and reduce the pathological damage of ischemic neurons.The mechanism may be related to inhibiting the activation of NLRP3 and pyroptosis pathway.
作者
王少华
孙凡凡
薛威
江勤
董六一
Wang Shaohua;Sun Fanfan;Xue Wei(Dept of Neurosurgery,Affiliated Fuyang Hospital of Anhui Medical University,Fuyang 236000;Dept of Pharmacology,Anhui Medical University,Hefei 230032)
出处
《安徽医科大学学报》
CAS
北大核心
2020年第9期1321-1326,共6页
Acta Universitatis Medicinalis Anhui
基金
安徽省自然科学基金(编号:1408085MH174)。
关键词
灯盏花素
慢性脑缺血
炎症小体
焦亡
凋亡
breviscapine
chronic cerebral ischemia
inflammasome
pyroptosis
apoptosis
作者简介
王少华,男,硕士,副主任医师;董六一,男,教授,博士生导师,责任作者,E-mail:dongly@ahmu.edu.cn。
