不同针刺手法时长对脑出血模型大鼠血脑屏障保护机制的研究

Protective Mechanism of Blood-brain Barrier of Cerebral Hemorrhage Model Rats with Different Acupuncture Manipulation Duration

目的:探究不同针刺手法时长对脑出血模型大鼠脑组织血脑屏障紧密连接蛋白的影响。方法:将20只SD雄性大鼠按照随机数字表法分为假手术组、模型组、针刺5s组、针刺60s组,每组5只。脑出血模型采用尾状核自体血缓慢注入法,造模成功后,针刺5s组和针刺60s组针刺水沟穴及双侧内关穴,分别行针5s和60s,每天1次,共干预5d;假手术组及模型组不予针刺。5d后比较光镜下(400倍)大鼠脑血肿区紧密连接蛋白Occludin、Claudin-5和JAM-1免疫荧光阳性细胞表达的情况。结果:①各组大鼠血肿区域Occludin免疫荧光阳性细胞表达比较,假手术组出现生理性表达;与假手术组比较,模型组、针刺5s组、针刺60s组Occludin表达均显著降低,差异具有统计学意义(P<0.05);与模型组比较,针刺5s组、针刺60s组Occludin均升高,且针刺60s组升高更显著,差异具有统计学意义(P<0.05)。②各组大鼠血肿区域Claudin-5免疫荧光阳性细胞表达比较,假手术组Claudin-5出现生理性表达;与假手术组比较,模型组、针刺5s组、针刺60s组Claudin-5表达均显著降低,差异具有统计学意义(P<0.05);与模型组比较,针刺5s组、针刺60s组Claudin-5均升高(P<0.05)。③各组大鼠血肿区域JAM-1免疫荧光阳性细胞表达比较,假手术组JAM-1呈生理性表达;与假手术组比较,模型组、针刺5s组、针刺60s组JAM-1表达均显著降低,差异具有统计学意义(P<0.05);与模型组比较,针刺5s组表达不显著,差异无统计学意义(P>0.05);与模型组比较,针刺60s组JAM-1升高显著,差异具有统计学意义(P<0.05)。结论:较长时间针刺手法对脑出血后血脑屏障的保护有正向作用,可增加血脑屏障的通透性。

Objective: To explore the effect of different acupuncture manipulation duration on the blood-brain barrier tight junction protein in rats with cerebral hemorrhage. Methods: A total of twenty SD male rats were divided into sham-operated group, model group, acupuncture 5 s group and acupuncture 60 s group by random number table method, with 5 rats in each group. Caudate nucleus autologous blood injection was used in cerebral hemorrhage model. After the establishment of cerebral hemorrhage model, the acupuncture groups received acupuncture on "shuigou" point and bilateral "neiguan" points, and received acupuncture on 5 s or 60 s every day. The sham-operated and model groups did not receive acupuncture. Five days later, the expressions of Occludin, Claudin-5 and JAM-1 immunofluorescence positive cells in the cerebral hematoma area of rats under light microscope(×400) were compared. Results:(1) Expression of Occludin immunofluorescence positive cells: Compared with the sham-operated group, the Occludin expression in the model, acupuncture 5 s and 60 s groups decreased significantly(P<0.05). Compared with the model group, the Occludin increased both in the acupuncture 5 s and 60 s groups. The increase was more significant in the 60 s acupuncture group(P<0.05);(2) Expression of Claudin-5 immunofluorescence positive cells: Claudin-5 physiological expression was in the sham-operated group. Compared with the sham-operated group, the Claudin-5 expression in the model group, acupuncture 5 s and 60 s groups were significantly reduced(P<0.05). Compared with the model group, Claudin-5 in the 5 s and 60 s acupuncture groups increased(P<0.05);(3) Expression of JAM-1 immunofluorescence positive cells: JAM-1 was physiologically expressed in the sham-operated group. Compared with the sham-operated group, the expression of JAM-1 in the model, acupuncture 5 s and 60 s groups were significantly decreased( P<0.05). Compared with the model group, the expression of the acupuncture 5 s group was not significant(P>0.05). The JAM-1 level in the acupuncture 60 s group increased significantly(P<0.05). Conclusion: Long-time acupuncture has a positive effect on the protection of the blood-brain barrier after cerebral hemorrhage and can increase the permeability of the blood-brain barrier.