摘要
目的基于p38丝裂原活化蛋白激酶(mitogen-activated protein kinase,MAPK)通路探究艾灸督脉组穴对血管性痴呆(vascular dementia,VD)大鼠的脑保护机制。方法将60只SPF级健康雄性SD大鼠随机分为假手术组(12只)和实验组(48只);实验组大鼠采用双侧颈总动脉永久结扎法复制VD大鼠模型,随后将模型复制成功的大鼠分为模型组、艾灸组、阻滞剂组、艾灸+阻滞剂组,每组12只;艾灸组大鼠取“百会”“大椎”“风府”穴,采用温和灸法治疗,每次30 min,每日1次,治疗6 d,休息1 d,连续治疗4周;阻滞剂组在模型复制前30 min腹腔注射p38 MAPK阻滞剂(10μmol/L SB203580),模型复制成功后隔日注射1次,每次每只大鼠按1 mL/kg注射10μmol/L SB203580,连续注射4周;艾灸+阻滞剂组大鼠在阻滞剂组干预方法基础上进行艾灸治疗。采用Morris水迷宫实验检测大鼠认知功能,TUNEL法检测大鼠海马组织CA1区神经元凋亡率,Western blot法和RT-qPCR法分别检测大鼠海马组织CA1区p38 MAPK、Bcl-2、Caspase-3、Caspase-9蛋白及mRNA表达水平。结果与模型组比较,艾灸组、阻滞剂组、艾灸+阻滞剂组大鼠平均逃避潜伏期均显著缩短(P<0.05),穿越平台次数显著增加(P<0.05),海马组织CA1区神经元凋亡率显著降低(P<0.05),p38 MAPK、Caspase-3、Caspase-9蛋白及mRNA表达水平均显著降低(P<0.05),Bcl-2蛋白及mRNA表达水平均显著升高(P<0.05);艾灸+阻滞剂组大鼠平均逃避潜伏期较艾灸组、阻滞剂组显著缩短(P<0.05),穿越平台次数显著增加(P<0.05),海马组织CA1区神经元凋亡率显著降低(P<0.05),p38 MAPK、Caspase-3、Caspase-9蛋白及mRNA表达水平均显著降低(P<0.05),Bcl-2蛋白及mRNA表达水平均显著升高(P<0.05)。结论艾灸督脉组穴对VD大鼠的认知功能具有一定的改善作用,其作用机制可能与调控p38 MAPK信号通路抑制海马神经元凋亡有关。
Objective To investigate the cerebral protection mechanism of governor vessel moxibustion in rats with vascular dementia(VD)based on the p38 mitogen-activated protein kinase(MAPK)pathway.Methods A total of 60 specific pathogen-free healthy male Sprague-Dawley rats were randomly divided into sham operation group with 12 rats and experimental group with 48 rats.The rats in the experimental group were treated with permanent ligation of both common carotid arteries to establish a rat model of VD,and after successful modeling,the rats were randomly divided into model group,moxibustion group,inhibitor group,and moxibustion+inhibitor group,with 12 rats in each group.The rats in the moxibustion group were given mild-warm moxibustion at Baihui,Dazhui,and Fengfu points for 30 minutes each time,once a day for 6 days,followed by 1 day of rest,for 4 consecutive weeks;the rats in the inhibitor group were given intraperitoneal injection of p38 MAPK inhibitor(10μmol/L SB203580)at 30 minutes before modeling,followed by the injection of 10μmol/L SB203580 every other day after modeling at a dose of 1 mL/kg for 4 consecutive weeks;the rats in the moxibustion+inhibitor group were given moxibustion in addition to the intervention in the inhibitor group.The Morris water maze test was used to observe the cognitive function of rats;the TUNEL method was used to measure the apoptosis rate of neurons in the hippocampal CA1 region;Western blot and RT-qPCR were used to measure the protein and mRNA expression levels of p38 MAPK,B-cell lymphoma-2(Bcl-2),Caspase-3,and Caspase-9 in the hippocampus.Results Compared with the model group,the moxibustion group,the inhibitor group,and the moxibustion+inhibitor group had a significantly shorter mean escape latency(P<0.05),a significant increase in the number of platform crossings(P<0.05),and a significant reduction in the apoptosis rate of neurons in the hippocampal CA1 region(P<0.05),as well as significant reductions in the protein and mRNA expression levels of p38 MAPK,Caspase-3,and Caspase-9(P<0.05)and significant increases in the protein and mRNA expression levels of Bcl-2(P<0.05).Compared with the moxibustion group and the inhibitor group,the moxibustion+inhibitor group had a significantly shorter escape latency(P<0.05),a significant increase in the number of platform crossings(P<0.05),and a significant reduction in the apoptosis rate of neurons in the hippocampal CA1 region(P<0.05),as well as significant reductions in the protein and mRNA expression levels of p38 MAPK,Caspase-3,and Caspase-9(P<0.05)and significant increases in the protein and mRNA expression levels of Bcl-2(P<0.05).Conclusion Governor vessel moxibustion has a certain effect in improving cognitive impairment in rats with VD,possibly by regulating the p38 MAPK signaling pathway and inhibiting the apoptosis of hippocampal neurons.
作者
刘芳
李梦醒
李真
王玉
唐巍
LIU Fang;LI Mengxing;LI Zhen;WANG Yu;TANG Wei(School of Medicine,Anqing Medical and Pharmaceutical College,Anhui Anqing246052,China;College of Acupuncture and Moxibustion,Anhui University of Chinese Medicine,Anhui Hefei230012,China;College of Physical Education,Anqing Normal University,Anhui Anqing246113,China)
出处
《安徽中医药大学学报》
CAS
2024年第2期42-48,共7页
Journal of Anhui University of Chinese Medicine
基金
安徽高校自然科学研究重点项目(2023AH053429,KJ2020A0377)
安徽高校优秀青年人才支持项目(gxyqZD2022109)。
