摘要
目的:探讨三焦针法对SAM小鼠血脑屏障(BBB)通透性及RhoA/Rho相关蛋白激酶(ROCK)的影响,并阐明其作用机制。方法:36只SAMP8老年痴呆小鼠随机分为模型组、非穴位组(腹中线平行、过髂嵴尖端直上2 mm处,1.5寸毫针向下斜刺约3 mm,角度90°~180°、频率60~120 min^(-1)、每侧各105 s)、穴位组[血海穴(双)直刺2~3 mm,大幅度(角度>180°)、低频率(频率<60 min^(-1))捻转泻法30 s,膻中、中脘、气海、足三里(双)直刺2~3 mm,小幅度(角度<90°)、高频率(频率>120 min^(-1))捻转泻法30 s],每组12只;12只SAMR1正常衰老小鼠作为对照组。采用Morris水迷宫实验评估各组小鼠学习和记忆功能,伊文思蓝染色法检测各组小鼠脑组织中伊文思蓝含量(代表BBB通透性),电镜下观察各组小鼠BBB超微结构,蛋白免疫印迹检测各组小鼠海马组织中闭合蛋白(Occludin)、密封蛋白5(Claudin-5)、闭锁连接蛋白1(ZO-1)、RhoA和ROCK蛋白表达水平。结果:模型组和非穴位组小鼠血管内皮细胞边界模糊,基底膜增厚,部分出现断裂现象,血管水肿明显;穴位组小鼠血管周围水肿现象明显缓解,内皮细胞基本处于正常状态,BBB情况得以改善。与对照组比较,模型组小鼠逃避潜伏期、脑组织中伊文思蓝含量和海马组织中RhoA和ROCK蛋白表达水平升高(P<0.05),海马组织中Occludin、Claudin-5和ZO-1蛋白表达水平降低(P<0.05),空间探索时间减少(P<0.05);模型组与非穴位组上述指标比较差异无统计学意义(P>0.05);与非穴位组比较,穴位组小鼠逃避潜伏期、脑组织中伊文思蓝含量和海马组织中RhoA和ROCK蛋白表达水平降低(P<0.05),海马组织中Occludin、Claudin-5和ZO-1蛋白表达水平升高(P<0.05),空间探索时间增加(P<0.05)。结论:三焦针法可以改善SAM小鼠BBB通透性,其机制可能与抑制RhoA/ROCK通路有关。
Objective:To investigate the effect of Sanjiao acupuncture on the permeability of the blood brain barrier(BBB)of the SAM mice and RhoA/Rho associated protein kinase(ROCK),and to clarify its mechanism.Methods:A tolal of 36 SAMP8 senile dementia mice were randomly divided into model group,non-acupoint group(parallel to the midline of the abdomen,inserted the needle 2 mm above the tip of the iliac crest,used a 1.5-inch needle to obliquely puncture about 3 mm downward,with angle of 90°-180°,frequency of 60-120 min^(-1),and 105 s on each side),acupoint group[Xuehai point(double)straight puncture 2-3 mm,large amplitude(angle>180°),low frequency(frequency<60 min^(-1))twisting and purging method for 30 s,Tanzhong,Zhongwan,Qihai,Zusanli(Double)Direct puncture 2-3 mm,small amplitude(angle<90°),high frequency(frequency>120 min^(-1))twirling and purging for 30 s],12 mice per group;12 SAMR1 normal aging mice were set as controls group.Morris water maze experiment was used to evaluate the learning and memory function of the mice;Evans blue staining method was used to detect the contents of Evans blue in the brain tissue of the mice in various groups(representing the BBB permeability),and the BBB ultrastructures of the mice in various groups were observed under electron microscope;Western blotting method was performed to detect the expression levels of Occludin,Claudin-5,zonula occludens-1(ZO-1),RhoA,ROCK proteins in hippocampus tissue of the mice in various groups.Results:In model group and non-acupoint group,the boundary of vascular endothelial cells of the mice was blurred,the basement membrane was thickened,some of them were broken,and the vascular edema was obvious;in acupoint group,the perivascular edema of the mice was significantly relieved,the endothelial cells were basically in normal state,and BBB was improved.Compared with control group,the escape latency of the mice,the content of Evans blue in the brain tissue,and the expression levels of RhoA and ROCK proteins in the hippocampus tissue were increased(P<0.05);
作者
王煜
阚伯红
赵岚
史慧妍
贾玉洁
WANG Yu;KAN Bohong;ZHAO Lan;SHI Huiyan;JIA Yujie(Department of Acupuncture,First Teaching Hospital,Tianjin University of Traditional Chinese Medicine,Tianjin 300193,China;National Clinical Research Center for Chinese Medicine Acupuncture and Moxibustion,Tianjin 300193,China;Institute of Acupuncture and Moxibustion,First Teaching Hospital,Tianjin University of Traditional Chinese Medicine,Tianjin 300193,China)
出处
《吉林大学学报（医学版）》
CAS
CSCD
北大核心
2021年第5期1086-1091,共6页
Journal of Jilin University：Medicine Edition
基金
国家自然科学基金项目(81603686)
国家自然科学基金青年科学基金项目(81704148)
天津市教委科研计划项目(2017KJ149)。
