摘要
目的观察电针对荷乳腺癌小鼠视交叉上核(suprachiasmatic nucleus,SCN)内Period基因的2个亚型Per1、Per2mRNA的表达及蛋白含量的影响,探讨荷乳腺癌小鼠电针后自发性活动节律变化的分子调控机制。方法将18只C57BL/6J雄性小鼠随机分为空白组、模型组、电针组3组,每组6只。LD驯化10d后关闭光源,使小鼠处于自由运行状态(DD状态)。DD驯化10d后将含有MA-782小鼠乳腺癌细胞株的混悬液接种于模型组及电针组小鼠左侧腋下,建立荷瘤小鼠模型,成瘤后继续监测节律。7d后电针组小鼠进行电针干预,在CT12时相点针刺百会穴和长强穴,连续电针3d;空白组和模型组小鼠均在相同时相点CT12采用相同的方法捆绑固定,不予电针刺激。第3次电针后2h处死动物,切取脑部SCN组织,采用实时荧光定量RT-PCR法测定各组小鼠SCN内Per1、Per2mRNA表达量,采用Western Blot法测定SCN内PER1、PER2蛋白含量。结果荷瘤小鼠SCN内Per1、Per2mRNA表达水平与空白组相比有明显降低趋势,无统计学意义(P>0.05),电针后,mRNA表达量较模型组有明显升高趋势,尤以Per2的差异明显(P<0.05)。荷瘤小鼠蛋白表达量与空白组及电针组相比均有明显差异,其差异具有统计学意义(P<0.05)。结论在CT12时相点电针百会和长强穴可以升高荷乳腺癌小鼠SCN内Per1、Per2mRNA的表达水平及其蛋白含量,提示电针对SCN内Per1、Per2等基因的作用,可能更多的是在转录后的翻译水平上,因为蛋白是最终执行功能的分子,这可能是电针调整荷瘤动物节律的重要机制之一。
OBJECTIVE To observe the effects of electro-acupuncture on mRNA expression of two subtypes of Period genes -Per1 and Per2 and protein contents in suprachiasmatic nucleus （SCN） of mice bearing breast cancer and explore the regulating mechanism of mice＇s spontaneous activity rhythm after electro-acupuncture. METHODS Eighteen male C57BL/6J mice were randomly divided into 3 groups with 6 in each group： blank control group, model group and electro-acupuncture group. Each group received LD domestication for 10 days. LD was turned off so that the mice were in free movements （DD state） 10d after LD domestication. Then bearing tumor models were established by making mixed suspension of MA-782 breast cancer cell lines inoculate into left armpits of mice in model group and electro-acupuncture group 10d after DD domestication. The spontaneous activity rhythm of mice were monitored upon successful model establishment. Seven days later, electro-acupuncture group were treated with electro-acupuncture by acupuncturing Changqiang （GV1） and Baihui （GV20） for 3d. Meanwhile, blank group and model group were tied up on two acupoints at the same CT12 instead of electro-acupuncture. Mice were sacrificed 2h after the third electro-acupuncture stimulation. The RNA expression levels of Per1 and Per2 in SCN were detected by real-time PCR, and the protein contents were tested by Western Blotting. RESULTS The mRNA levels of Per1 and Per2 in SCN of model group were lower than blank group, but there was no statistical significance （P〈0.05）, while the levels were higher than model group, especially in Per2. There were significant statistical differences （P〈0.05）. The levels of protein in mice bearing breast cancer differed from blank group and electro-acupuncture group, and the difference was statistically significant （P〈0. 05）. CONCLUSION Electro-acupuncture on Changqiang （GV1） and Baihui （GV20） at CT12 may increase mRNA expression levels of Per1 and Per2 and their protein contents in SCN of mice b
出处
《南京中医药大学学报》
CAS
CSCD
北大核心
2014年第3期263-266,共4页
Journal of Nanjing University of Traditional Chinese Medicine
基金
国家自然科学基金(30873297)
