穿心莲内酯对慢性低灌注所致脑损伤的保护作用及其机制  

Protective effect of andrographolide on brain injury caused by chronic hypoperfusion and its mechanism

作　　者：宋糖 覃振敏 陈永欣 陈耀 周静 SONG Tang;QIN Zhenmin;CHEN Yongxin;CHEN Yao;ZHOU Jing(Teaching and Research Section of Physiology,School of Basic Medicine,Guangxi Medical University,Nanning 530021,Guangxi,China;Key Laboratory of Basic Research on Regional Diseases,Department of Education of Guangxi Zhuang Autonomous Region,Nanning 530021,Guangxi,China)

机构地区：[1]广西医科大学基础医学院生理学教研室,广西南宁市530021 [2]广西壮族自治区教育厅区域疾病基础研究重点实验室(广西医科大学),广西南宁市530021

出　　处：《广西医学》2024年第11期1705-1715,共11页Guangxi Medical Journal

基　　金：国家自然科学基金(32160160)。

摘　　要：目的探讨穿心莲内酯对慢性低灌注所致脑损伤的治疗效果及其作用机制。方法(1)动物实验。将40只SD大鼠按随机数字表法分为假手术组、造模组、低剂量组、中剂量组和高剂量组,每组8只。对造模组和3个不同剂量组大鼠采用双侧颈总动脉结扎法永久结扎双侧颈总动脉,假手术组仅行假手术。造模后28 d,给予低剂量组大鼠灌胃20 mg/kg穿心莲内酯,中剂量组大鼠灌胃40 mg/kg穿心莲内酯,高剂量组大鼠灌胃80 mg/kg穿心莲内酯,假手术组和造模组灌胃等体积生理盐水,1次/d,持续14 d。(2)细胞实验。将细胞分为对照组、造模组、1μmol/L穿心莲内酯组、2μmol/L穿心莲内酯组和4μmol/L穿心莲内酯组。各穿心莲内酯干预组给予相应浓度的穿心莲内酯干预,对照组、造模组加入等体积完全培养基。干预24 h后,除对照组外,将其余4组细胞置入密闭的缺氧小室中进行氧糖剥夺法造模6 h。(3)通过水迷宫和旷场实验进行动物行为学检测评估大鼠的学习记忆功能;取大鼠脑组织进行HE染色和Nissl染色观察脑组织神经细胞的病理状态;采用ELISA试剂盒检测大鼠脑组织炎症因子表达量;使用CCK⁃8和Hoechst染色检测并观察HT22细胞活性状态;采用Western blot检测细胞和大鼠脑组织核因子κB(NF⁃κB)通路相关蛋白的表达。结果(1)动物实验。与假手术组相比,造模组逃避潜伏期和第1次穿越平台时间增加,穿越平台次数、直立次数和穿越格子数减少;与造模组相比,3个剂量组大鼠逃避潜伏期、第1次穿越平台时间缩短,穿越平台次数增加(P<0.05)。造模组大鼠海马区和皮质区神经元排列紊乱,细胞质萎缩,核仁碎裂;3个剂量组大鼠海马区和皮质区神经元损伤程度较造模组减轻。与假手术组相比,造模组大鼠海马区和皮质区神经元内Nissl小体明显减少,神经元受损;3个剂量组大鼠神经元内Nissl小体数量较造模组增加,损伤�Objective To investigate the therapeutic effect of andrographolide on brain injury caused by chronic hypoperfusion and its mechanism.Methods(1)Animal experiment.A total of 40 SD rats were assigned to sham operation group,modeling group,or low⁃,medium⁃,and high⁃dose groups according to the random number table method,with 8 rats in each group.Rats of the modeling and 3 different doses groups received permanent bilateral common carotid artery occlusion by bilateral common carotid artery occlusion method,while the sham operation group only underwent sham operation.After 28 days of modeling,the low⁃,medium⁃,and high⁃dose groups received intragastric administrations of andrographolide in 20 mg/kg,40 mg/kg and 80 mg/kg,respectively,whereas the sham operation and modeling groups received intragastric administration of normal saline in equivalent volume,once a day,for 14 days continuously.(2)Cell experiment.Cells were assigned to control group,modeling group,1μmol/L andrographolide group,2μmol/L andrographolide group,or 4μmol/L andrographolide group.Various andrographolide intervention groups received andrographolide in corresponding concentrations for intervention,while equivalent volume complete medium was added to the control and modeling groups.After 24⁃hour intervention,except for the control group,cells of the remaining 4 groups were placed in a closed anoxic chamber and the model was established by oxygen⁃glucose deprivation for 6 hours.(3)Ethology was detected and learning memory function of rats was evaluated by water maze and open field test.The pathological states of nerve cells in brain tissues were observed by performing the HE and Nissl staining on brain tissues of rats.The ELISA kit was employed to detect expressions of inflammatory factors in brain tissues.The CCK⁃8 and Hoechst staining were used to detect and observe HT22 cell active state.The Western blot was employed to detect expressions of cell and proteins related to nuclear factor kappa B(NF⁃κB)pathway in brain tissues of rat

关 键 词：穿心莲内酯 慢性脑低灌注 血管性认知障碍 核因子ΚB 炎症因子 大鼠 

分 类 号：R285.5[医药卫生—中药学]