井穴刺络放血对脑缺血再灌注小鼠线粒体生物合成的影响

Effects of Bloodletting at Jing-Well Points on Mitochondrial Biogenesis in Mice with Cerebral Ischemia Reperfusion

目的:探讨井穴刺络放血对脑缺血再灌注小鼠脑水肿及线粒体生物合成的影响。方法:将36只C57BL/6雌性小鼠随机均分对照组、脑缺血再灌注组和井穴刺络放血组,脑缺血再灌注组和井穴刺络放血组采用线栓法构建大脑中动脉栓塞后再灌注模型,造模后井穴刺络放血组用1 mL注射器针头刺破小鼠双侧前肢趾端井穴位放血0. 5μL,1次/12 h,共6次;于术后第72 h时对3组小鼠进行改良神经功能损伤评分(mNSS)后处死小鼠,留取5 mg同一部位脑组织;用real-time PCR法检测留取脑组织中PGC-1α、NRF1、PPARδ、TFAM基因表达水平和线粒体DNA拷贝数,剩余的脑组织用于测定脑组织含水量。结果:井穴刺络放血组和脑缺血再灌注组神经功能损伤评分及脑水肿程度高于对照组,且脑缺血再灌注组高于井穴刺络放血组,差异有统计学意义(P <0. 05);井穴刺络放血组和脑缺血再灌注组小鼠脑组织中PGC-1α、TFAM和PPARδ基因表达以及线粒体DNA拷贝数均高于对照组,且井穴刺络放血组高于脑缺血再灌注组,差异有统计学意义(P <0. 05)。结论:井穴刺络放血可能通过促进线粒体生物合成,使脑水肿程度减轻并发挥脑保护作用。

Objective：To investigate the impact of bloodletting from well points on brain edema and mitochondrial biosynthesis in mice with cerebral ischemia-reperfusion. Methods： 36 C57BL/6 female mice were randomly divided into three groups, including control group, cerebral ischemia-reperfusion group and blood-letting puncture in ＂Twelve-Well Points＂ group. The latter two groups used suture-occluded method to build a model of reperfusion after the first construct of middle cerebral artery （MCA） embolism. After the model was established, the blood-letting group used a l mL syringe needle to puncture 0.5 μL of blood from the acupuncture points of the forelimbs of mice once every 12 hours and for a total of 6 times. 72 hours after operation, the mice were sacrificed after modified neurological severity score （mNSS）, and 5 mg of brain tissue was taken from the same site. Real-time PCR was used to detect PGC-1α, NRF1, PPARδ and TFAM gene expression levels in brain tissue and mitochondrial DNA copy number and the remaining brain tissue was used to determine brain water content. Methods： The neurological impairment score and water content of brain tissue in bloodletting group and reperfusion group was significantly increased when compared with control group, and reperfusion group was higher than that in bloodletting group, the difference was statistically significant （ P 〈0.05）. PGC-1α, TFAM and PPARδ gene expression and mitochondrial DNA copy number in bloodletting group and reperfusion group were significantly increased than that of control group, and bloodletting group increased more than that in reperfusion group （ P 〈0.05）.