针刺太冲、太溪对自发性高血压大鼠下丘脑病理形态的影响

Effect of Acupuncture at Taichong and Taixi on Pathological Morphology of Hypothalamus in Spontaneously Hypertensive Rats

目的:通过观察针刺太冲、太溪后自发性高血压大鼠的下丘脑HE染色、尼氏染色及两者的半定量等指标。探讨课题前期功能磁共振结果在分子生物学层面的机制。最终,尝试寻求针刺对自发性高血压大鼠的降压机制,为阐明针灸降低血压的原理提供理论和实验依据。方法:采用STATA 14.0统计软件以原始血压为分层条件按照随机区组设计将SHRs随机分成3组:模型组、冲溪组、非穴组,每组12只。WKY大鼠不分组。针灸处方:太冲(双)、太溪(双)、非穴(双)。操作方法:在大鼠清醒状态下行针刺捻转手法,平补平泻,于每天上午8∶30开始进行,左右两侧穴位同时进行针刺,直刺,先左侧穴位行针2.5 min,然后右侧穴位行针2.5 min,每次持续总共时间5 min,针刺每日1次,逢周日休息1 d,连续针刺4周,共针刺24次。捻转频率为(80±5)次/min,捻转幅度(180±5)°。所有针刺操作由固定一人进行。正常组与模型组不给予针刺,抓取固定5 min。采用HE染色和尼氏染色观察大鼠下丘脑的一般病理形态结构;采用ImageJ对细胞数量进行半定量计算。结果:各组大鼠下丘脑HE染色及尼氏染色后所得病理结果与同脑区正常组相比较,未观察到明显的病理变化。但相较于正常组,其余各组细胞总数及尼氏体数量均有不同程度减少。通过半定量方式进行证实。细胞总数及尼氏体数量从多到少依次是:正常组、冲溪组、非穴组、模型组。结论:针刺太冲、太溪对SHRs降压作用在病理形态学上的可能机制在于减少或阻止下丘脑的细胞减少和凋亡,解除细胞增殖能力的抑制,对N-甲基-D-天冬氨酸(NMDA)受体信号传导的调节,调整下丘脑室旁核肾上腺髓质素含量水平、调节Toll样受体4(Toll-like receptor 4,TLR4)所参与的促炎细胞因子生成及脑源性神经营养因子(BDNF)的含量水平而实现下丘脑的保护。最终通过上述多种机制,而使得一系列与下丘脑相关的血压调控途径得到调整,实现降压目的。

Objective:To observe the hypothalamic HE staining and Nissl staining of spontaneously hypertensive rats after acupuncture at Taichong(LR3)and Taixi(KI3).To discuss topics preliminary results of functional magnetic resonance mechanism in molecular biology level and try to seek the antihypertensive mechanism of acupuncture on spontaneously hypertensive ratsand provide the theoretical and experimental basis for elucidating the principle of acupuncture to lower blood pressure.Methods:By STAT 14.0,the SHRs were randomly divided into three groups according to the randomized block design using the original blood pressure as the stratification condition:Model group,Chongxi group and Sham acupuncture group,12 in each group.WKY rats were not grouped.Acupuncture prescriptions:Tai Chong(double,LR3),Taixi double,KI3),non-acupoint(double).Method of operation:Acupuncture in the awake state of the rats by gentle tonifying and purging manipulation,starting at 8∶30 everyday.Both sides were needled at the same time.First the left side was needled for 2.5 mins and then the the right side was needled for 2.5 mins,totally lasting for 5 mins,once a day.There was a rest on Sunday.The acupuncture was lasted for 4 weeks,a total of 24 times.The twisting frequency was(80±5)times/min,and the twisting amplitude was(180±5)°.All acupuncture operations were performed by the same person.The normal group and the model group were not given acupuncture,and the grasping was fixed for 5 min.The general pathological morphology of the rat hypothalamus was observed by HE staining and Nissl staining.The number of cells was semi-quantitatively calculated using ImageJ.Results:HE staining and hypothalamus of rats after Nissl pathology results obtained the same brain regions as compared to the normal group and no significant pathological changes were observed.However,compared with the normal group,the total number of cells and the number of Nissl bodies in the other groups were reduced to some extent.Confirmed by semi-quantitative means,the total number of cells and the number of Nissl bodies from high to low were normal group,Chongxi group,Sham acupuncture group and model group.Conclusion:The possible mechanism of acupuncture at Taichong(LR3)and Taixi(KI3)on the pathological morphology of antihypertensive effect of SHRs is to reduce or prevent cell reduction and apoptosis of hypothalamus,and abolish the inhibition of cell proliferation,for N-methyl-D-Regulation of aspartate(NMDA)receptor signaling,regulation of hypothalamic paraventricular nucleus adrenomedullin levels,regulation of pro-inflammatory cytokines involved in Toll-like receptor 4(TLR4).The level of brain-derived neurotrophic factor(BDNF)is produced to achieve protection of the hypothalamus.Finally,through a variety of mechanisms described above,a series of blood pressure regulation pathways related to the hypothalamus are adjusted to achieve the purpose of blood pressure reduction.