“百会”透“曲鬓”针刺法对脑出血大鼠脑组织中PERK蛋白表达影响的实验研究

摘要

目的:探讨“百会”透“曲鬓”针刺法对脑出血大鼠脑组织细胞内质网应激PERK通路关键蛋白PERK表达的影响.方法:将Wistar雄性大鼠随机分为SHAM组、模型组、针刺组、3-MA组、3-MA+针刺组,再将各组随机分为4个时间亚组,每亚组6只.采用自体血注射的方法建立脑出血大鼠模型,3-MA组于造模前15 min注射3-MA自噬抑制剂,针刺组和3-MA+针刺组于造模成功后以“百会”透“曲鬓”针刺法进行干预,于相应时间节点取材.根据改良版神经缺损评分系统(mNSS)对大鼠神经功能进行评价,并用Western Blot法分析PERK蛋白相对表达量的变化.结果:神经功能评价方面,造模前大鼠无神经功能缺损体征.造模后SHAM组大鼠各时间点评分无显著差异,其余各组神经功能缺损体征于72 h时各项缺损体征达到最高峰,后有所缓解,7d时针刺组神经功能缺损体征明显改善(P＜0.05),3-MA组于各时间点神经功能缺损最重,3-MA+针刺.组神经缺损情况较3-MA组轻.West-ern Blot法检测结果显示,除SHAM组外,其余各组PERK蛋白相对表达量均有随时间变化的趋势,分别于6h最低,后逐渐升高,至7d最高;相同时间点,针刺组相对表达量最高(P＜0.01),3-MA组蛋白相对表达量明显低于其他各组(P＜0.05),3-MA+针刺组表达量略高于3-MA组.结论:“百会”透“曲鬓”针刺法对脑出血大鼠神经功能恢复具有促进作用,可能促进内质网应激PERK通路的激活.%Objective:To investigate the effect of Baihui-Qubin acupuncture on PERK expression of PERK pathway key proteins in intracerebral hemorrhage (ICH) rat models.Methods:Wistar rats were randomly divided into five groups:sham group,ICH model group,inhibitor group,acupuncture group and inhibitor + acupuncture group (24 rats in each group);4 sub-groups were divided based on different time points (12hrs,24hrs,72hrs,7days),with 6 rats in each subgroup.Establish intracerebral hemorrhage rat models by autologous blood injection.Brain tissues were harvested at altered time points in sham group and ICH model group.3-MA inhibitor was injected before 15 minutes of autologous blood injection.Baihui-Qubin acupuncture was implemented in acupuncture group and inhibitor + acupuncture group from 6hrs after ICH (once per 24hrs).Brain tissues were also harvested at altered time points.Evaluate the neurological deficit at 6hrs after ICH and before tissues being harvested.Western Blot studies were performed to analyze the expression of PERK in brain tissues.Results:Neurological deficit evaluation:the neurological behave in acupuncture group was improved,which was associated with the acupuncture involvement.Western Blot results showed that the expression of PERK was increased in acupuncture group and reduced in inhibitor group as well as the inhibitor + acupuncture group which was slightly higher than that in the inhibitor group.Conclusion:Baihui-Qubin acupuncture therapy can alleviate the neurological deficits and also has an effect on PERK pathway activation.