摘要
目的:探讨好忘方对血管性痴呆(VD)大鼠学习记忆障碍的影响及机制。方法:70只雄性SD大鼠适应性喂养1周后,随机抽选10只作为假手术组,仅分离颈总动脉不结扎,剩余大鼠将采用颈总动脉分次结扎法制作VD大鼠模型。剔除死亡和造模不成功的大鼠后,将造模成功的大鼠随机分为模型组、盐酸多奈哌齐组(0.45 mg/kg)、好忘方低剂量组(3.96 g/kg)、好忘方中剂量组(7.92 g/kg)、好忘方高剂量组(15.84 g/kg),每组10只,各组均给予相应药物灌胃治疗4周后取材。Morris水迷宫实验检测各组逃避潜伏期、跨越平台次数、苏木素-伊红(HE)染色检测海马神经元损伤状况;TUNEL染色法观察细胞凋亡;ELISA检测大鼠海马组织炎症因子含量;生化试剂盒检测海马组织中超氧化物歧化酶(SOD)、谷胱甘肽过氧化物酶(GSH-Px)和丙二醛(MDA)含量;RT-PCR测定各组大鼠海马组织中Caspase-3、Caspase-9、Bcl-2、Bax mRNA水平;Western blot测定相关蛋白的表达。结果:与假手术组比较,模型组大鼠逃避潜伏期增加(P<0.01),穿越平台次数下降(P<0.01),海马区神经元层次减少,数量骤减,结构排列不齐,IL-1β、IL-6和TNF-α含量升高(P<0.01),神经元细胞凋亡率显著升高(P<0.01),海马组织SOD、GSH-Px活性降低,MDA含量升高(P<0.01),海马组织Caspase-3、Caspase-9、Bax mRNA和蛋白表达升高,Bcl-2 mRNA和蛋白表达降低(P<0.01);与模型组比较,好忘方各剂量组、盐酸多奈哌齐组大鼠逃避潜伏期降低(P<0.05,P<0.01),穿越平台次数增加(P<0.05,P<0.01),HE染色显示,海马神经元细胞数量增多,形态正常,排列整齐,IL-1β、IL-6和TNF-α含量降低(P<0.05,P<0.01),神经元细胞凋亡率明显减少(P<0.05,P<0.01),海马组织SOD、GSH-Px活性上升,MDA含量下降(P<0.05,P<0.01),Caspase-3、Caspase-9、Bax mRNA和蛋白表达显著下降(P<0.05,P<0.01),Bcl-2 mRNA和蛋白表达明显上升(P<0.05,P<0.01)。结论:好忘方可通过抑制神经元凋亡、神经炎症和氧化应激改善VD大鼠的学习记忆障碍。
Objective:To investigate the effect and mechanism of Haowang Formula on learning and memory impairment in rats with vascular dementia(VD).Methods:Seventy male SD rats underwent adaptive feeding for one week before the study.Ten rats were randomly assigned to the sham surgery group,in which only the common carotid artery was isolated without ligation.The remaining rats were subjected to sequential ligation of the common carotid artery for the modeling of VD.Rats that died or underwent unsuccessful modeling were excluded.The successfully modeled rats were randomly assigned into the following groups(n=10 per group):model group,donepezil hydrochloride group(0.45 mg/kg),low-dose Haowang Formula group(3.96 g/kg),medium-dose Haowang Formula group(7.92 g/kg),high-dose Haowang Formula group(15.84 g/kg).Each group was given corresponding medication by gavage once daily for four weeks,after which the outcome analyses were conducted.The escape latency and cross-platform times in each group were detected by Morris water maze test.Hematoxylin-eosin(HE)staining was employed to detect the damage of hippocampal neurons.TUNEL staining was used to observe cell apoptosis;The levels of hippocampus inflammatory factors in rats were measured by ELISA.Biochemical kits were used to measure the levels of superoxide dismutase(SOD),glutathione peroxidase(GSH-Px)and malondialdehyde(MDA)in the hippocampal tissue.RTPCR was utilized to analyze Caspase-3,Caspase-9,Bcl-2,and Bax mRNA levels in hippocampal tissue.By Western blot assay,the expression of relevant protein was detected.Results:Compared with the sham surgery group,rats in the model group exhibited increase in escape latency(P<0.01),a decrease in the number of platform crossings(P<0.01).Furthermore,hippocampal neuronal layers in the model group were disorganized and reduced in number(P<0.01).In addition,the contents of IL-1β,IL-6 and TNF-αincreased(P<0.01),neuronal apoptosis rate increased(P<0.01),levels of SOD and GSH-Px in the hippocampal tissue decreased(P<0.01),contents of MDA increases,the expression of Caspase-3,Caspase-9,Bax mRNA and protein in hippocampus was increased,the expression of Bcl-2 mRNA and protein in hippocampus was decreased(P<0.01).Compared with the model group,rats in all Haowang Formula dose groups and donepezil hydrochloride shortened the escape latency(P<0.05,P<0.01)and increased the number of platform crossings(P<0.05,P<0.01).According to the HE staining,the number of hippocampal neuron cells increased with normal morphology and well-arranged alignment,the contents of IL-1β,IL-6 and TNF-αdecreased(P<0.05,P<0.01),the number of neuronal apoptosis decreased remarkably(P<0.05,P<0.01),the activities of SOD and GSH-Px in the hippocampal tissue increased,the content of MDA decreased(P<0.05,P<0.01).Moreover,it showed significantly reductions in Caspase-3,Caspase-9,Bax mRNA and protein expression(P<0.05,P<0.01),and significant increases in Bcl-2 mRNA and protein expression(P<0.05,P<0.01).Conclusion:Haowang Formula can improve learning and memory impairments in VD rats by inhibiting neuronal apoptosis,neuroinflammation and oxidative stress.
作者
张运辉
杨梦琳
李勇华
伍大华
杨昆
刘霞
程妍
ZHANG Yunhui;YANG Menglin;LI Yonghua;WU Dahua;YANG Kun;LIU Xia;CHENG Yan(Chongqing Three Gorges Medical College,Chongqing 404120,China;The Affiliated Hospital of Hunan Academy of Traditional Chinese Medicine,Changsha 410006,China)
出处
《中医药信息》
2026年第1期1-7,共7页
Information on Traditional Chinese Medicine
基金
国家自然科学基金项目(81874462)
重庆市中医药传承创新团队-三峡库区道地药材保护与利用多学科交叉创新团队(CXTD202303)
重庆市级乡村振兴对口帮扶科技项目(2023TIAD-ZXX0010)
重庆三峡医药高等专科学校呼吸和老年疾病中医药防治创新研究中心(2024年)
重庆三峡医药高等专科学校中医药防治呼吸和老年病团队项目(TIG202302)。
关键词
好忘方
血管性痴呆
学习记忆障碍
神经炎症
神经元凋亡
氧化应激
Haowang Formula
Vascular dementia
Learning and memory impairment
Neuroinflammation
Neuronal apoptosis
Oxidative stress
