针刺对VD模型大鼠脑组织细胞凋亡相关蛋白Caspase-3、NFK-βp65及PTEN mRNA表达水平的影响
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摘要
目的:探讨针刺对血管性痴呆(VD)大鼠脑组织中引起细胞凋亡相关蛋白Caspase-3、NFK-βp65及PTEN mRNA表达的影响,为临床治疗血管性痴呆提供理论依据。方法:将水迷宫测试合格的大鼠随机分5组(正常组、假手术组、模型组、手针组、电针组,每组24只)。运用四血管阻断法制作VD大鼠模型,造模成功7天后,针刺大鼠头穴干预。通过Morris水迷宫实验测试大鼠的学习记忆能力;免疫组化检测海马Caspase-3、NFK-βp65阳性表达。结果:相同时刻,针刺组(手针组、电针组)大鼠与模型组大鼠对比,成功穿越平台次数增多,逃避潜伏期缩短,差异有统计学意义(P <0. 05或P <0. 01);同一时间点,电针组大鼠较手针组穿越平台成功次数明显增多,差异有统计学意义(P <0. 05)。手针组和电针组治疗第7天、第14天、第21天3个时间点与同一时点模型组Caspase-3阳性细胞表达相比有所下降,均具有显著性差异(P <0. 05或P <0. 01),说明手针治疗和电针治疗均能降低Caspase-3阳性细胞表达。手针组和电针组治疗第7天、第14天、第21天3个时间点与同一时点模型组NFK-βp65阳性细胞表达相比有所下降,均具有显著性差异(P <0. 05或P <0. 01),说明手针治疗和电针治疗均能降低NFK-βp65阳性细胞表达。手针组和电针组治疗第7天、第14天、第21天3个时间点与同一时点模型组PTEN mRNA表达相比有所下降,均具有显著性差异(P <0. 01),说明手针治疗和电针治疗均能降低PTEN mRNA表达。结论:针刺具有明显抑制血管性痴呆(VD)大鼠脑组织细胞凋亡相关蛋白Caspase-3、NFK-βp65及PTEN mRNA表达的影响,具有脑保护作用。
Objective: To investigate the effect of acupuncture on the expressions of apoptosis-related proteins( caspase-3 and NFK-βp65) and PTEN mRNA in brain tissue of the rats with vascular dementia( VD),thus to provide theoretical basis for the clinical treatment of VD. Methods: 168 rats that passed the water maze test were randomly divided into five groups( the normal group,the sham operation group,the model group,the manual needling group,the electro-acupuncture group,24 rats in each group). The model of VD rats was made by four-blood vessel block method. 7 days after successful modeling,scalp needling intervention was applied. The learning ability and memory ability of the rats were measured by Morris water maze. The expressions of Caspase-3 and NFK-βp65 in hippocampus were detected by immunohistochemistry. Results: At the same time point,evasive incubation period was shortened,and the number of crossing platform was increased in the manual needling group and in the electro-acupuncture group compared to those in the model group( P < 0. 05,P < 0. 01); the number of crossing platform was significantly increased in the electro-acupuncture group compared to that in the the manual needling group( P < 0. 05). On 7 th day,14 th day,and 21 st day of the treatment,the expressions of Caspase-3 positive cells,NFK-βp65 positive cells,and PTEN mRNA were significantly reduced in the manual needling group and in the electro-acupuncture group compared to those in the model group( P < 0. 05,P < 0. 01). Conclusion: Acupuncture can effectively inhibit the expressions of caspase-3,NFK-βp65 and PTEN mRNA in brain tissue of rats with VD. It has brain protection.
Objective: To investigate the effect of acupuncture on the expressions of apoptosis-related proteins( caspase-3 and NFK-βp65) and PTEN mRNA in brain tissue of the rats with vascular dementia( VD),thus to provide theoretical basis for the clinical treatment of VD. Methods: 168 rats that passed the water maze test were randomly divided into five groups( the normal group,the sham operation group,the model group,the manual needling group,the electro-acupuncture group,24 rats in each group). The model of VD rats was made by four-blood vessel block method. 7 days after successful modeling,scalp needling intervention was applied. The learning ability and memory ability of the rats were measured by Morris water maze. The expressions of Caspase-3 and NFK-βp65 in hippocampus were detected by immunohistochemistry. Results: At the same time point,evasive incubation period was shortened,and the number of crossing platform was increased in the manual needling group and in the electro-acupuncture group compared to those in the model group( P < 0. 05,P < 0. 01); the number of crossing platform was significantly increased in the electro-acupuncture group compared to that in the the manual needling group( P < 0. 05). On 7 th day,14 th day,and 21 st day of the treatment,the expressions of Caspase-3 positive cells,NFK-βp65 positive cells,and PTEN mRNA were significantly reduced in the manual needling group and in the electro-acupuncture group compared to those in the model group( P < 0. 05,P < 0. 01). Conclusion: Acupuncture can effectively inhibit the expressions of caspase-3,NFK-βp65 and PTEN mRNA in brain tissue of rats with VD. It has brain protection.
引文
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[11]马春林,张宣,刘佳楠,等.归芪聪志汤对VD模型大鼠海马神经元Claudin-1,TGF-β表达的影响[J].中国实验方剂学杂志,2018,24(6):102-107.
[12]张鑫,李敏,彭晓燕,等.百会穴、大椎穴在血管性痴呆治疗中的作用机制探讨[J].辽宁中医杂志,2017,44(4):835-838.
[13]欧阳钢,田淳.四神聪穴的临床应用[J].针灸临床杂志,1998,14(5):41-42.
[14]裴蕾,戴晓矞.风池穴临床应用综述[J].新疆中医药,2014,32(2):84.
[2]郭明冬,李秋艳,韦云,等.戟天健脑方对血管性痴呆患者认知功能及胆碱能神经递质的影响[J].中国中医基础医学杂志,2018,24(9):1253-1255.
[3]陆连第,段伟松,赵玉,等.黄精多糖对血管性痴呆模型大鼠干预作用的实验研究[J].中药材,2018,41(9):1959-1962.
[4]张淑芳,吕光耀.中医药治疗血管性痴呆的研究进展[J].新疆中医药,2017,35(5):116-119.
[5] Schmidt-Kastner R,Paschen W,Ophoff BG,et al. A modified fourvesselocclusion model for inducing in complete forebrain ischemia inrats[J]. Stroke,1989,20(7):936.
[6]李忠仁.实验针灸学[M].北京:中国中医药出版社,2007:219-253.
[7]王绪平,呙登俊,陈宇,等. Morris水迷宫实验评价血管性痴呆大鼠模型的学习记忆能力[J].中华中医药学刊,2013,31(1):99-100.
[8]李欧,沙中玮,吴双,等.血管性认知功能障碍动物模型研究进展[J].中国中医基础医学杂志,2018,24(4):567-570.
[9]李君,姚刚,吴杰.血管性痴呆动物模型研究概况[J].中国医药导报,2014,11(14):157-160.
[10]曹乐,毕炳添,宾娟,等.乌参醒脑方对大鼠脑缺血致血管性痴呆的神经保护作用[J].中国新药杂志,2012,21(19):2306-2310.
[11]马春林,张宣,刘佳楠,等.归芪聪志汤对VD模型大鼠海马神经元Claudin-1,TGF-β表达的影响[J].中国实验方剂学杂志,2018,24(6):102-107.
[12]张鑫,李敏,彭晓燕,等.百会穴、大椎穴在血管性痴呆治疗中的作用机制探讨[J].辽宁中医杂志,2017,44(4):835-838.
[13]欧阳钢,田淳.四神聪穴的临床应用[J].针灸临床杂志,1998,14(5):41-42.
[14]裴蕾,戴晓矞.风池穴临床应用综述[J].新疆中医药,2014,32(2):84.