基于赫依三穴研究蒙医灸疗对PCPA致失眠模型大鼠镇静催眠作用机理研究

It is a safe and effective method for the mongolian moxibustion therapy to the insomia. Previous studies show that:① It will improve the the daily life quality of the insomnia model rats. ② It will reduce the contents of NO and reduce the activity of NOS which are the important neurotransmitters in the central nervous system (CNS). It has closely correlated with the sleeping quality for the hypothalamus-pituitary-adrenal(HPA) axis. The regulatory factors which is related with the HPA axis can improve the sleep quality by adjusting the HPA axis. The regulatory factors includes: interleukin-1(IL-1)、interleukin-1(IL-6)、tumor necrosis factor-α(TNF-α)、5-hydroxytryptamine (5-HT) 、norepinephrine (NE)、dopamine(DA) and gamma-aminobutyric acid transporter(GAT-1). After the mongolian moxibustion therapy we observe the behavior changes of the insomnia model rats which was induced by parachloroamphetamine acid (PCPA). And then we observe the expressions of the regulatory factors in the brain stem which is related with the HPA axis by methods of Elisa, high efficiency liquid chromatography(HPLC) and PCR. We try to explore the effect and mechanisms of the mongolian moxibustion therapy to the insomia and find the compatibility rules of the He yi three acupoint by these ways. And so we can illuminate the mechanism of the mongolian moxibustion therapy to insomia and provide theoretical basis for clinical therapy of insomia.

蒙医灸疗治疗失眠安全、有效。前期研究表明：①灸疗可改善睡眠紊乱模型大鼠一般生活状态。②可降低中枢神经系统内重要的神经递质NO含量与NOS活性。然而下丘脑-垂体-肾上腺(HPA)轴的功能与睡眠状态密切相关， HPA轴的相关调节因子白介素1（IL-1）、白介素6（IL-6）、肿瘤坏死因子（TNF-α）、5-羟色胺（5-HT）、去甲肾上腺素（NE）、多巴胺（DA）、γ-氨基丁酸转运体（GAT-1）通过调节HPA轴可以改善睡眠。因此，本研究对对氯苯丙胺酸（PCPA）致失眠模型大鼠进行灸法干预，观察失眠大鼠行为学变化，运用Elisa、高效液相色谱、PCR方法，观察大鼠脑干组织中HPA轴相关调节因子的表达，从而探寻灸疗对失眠作用机理及赫依三穴组穴配伍规律，进一步明确灸疗的起效途径，为临床治疗失眠提供理论依据。

【研究背景】本课题组通过前期研究证实，灸疗可改善睡眠紊乱模型大鼠一般生活状态，延长模型大鼠对戊巴比妥钠睡眠时间，可降低中枢神经系统内重要的神经递质NO含量与NOS活性。.【主要研究内容】在前期研究基础上，本研究对对氯苯丙胺酸（PCPA）致失眠模型大鼠进行了灸法干预，以赫依三穴（顶会穴、赫依穴、黑白际穴）为切入点，将实验动物随机分为正常组、模型组、安定组、顶会穴组、赫依穴组、黑白际穴组、顶会穴与赫依穴组、顶会穴与黑白际穴组、赫依穴与黑白际穴组、三穴组等10组，每组10只。运用Elisa、高效液相色谱、PCR方法，检测大鼠脑干组织中HPA轴相关调节因子的表达。探寻蒙医灸疗法镇静催眠机制及赫依三穴组穴配伍规律。.【重要结果】本研究主要观察了与睡眠密切相关的HPA轴相关调节因子白介素1（IL-1）、白介素6（IL-6）、肿瘤坏死因子（TNF-α）、5-羟色胺（5-HT）、去甲肾上腺素（NE）、多巴胺（DA）、γ-氨基丁酸转运体（GAT-1）等七个指标。经统计，发现赫依三穴在单穴治疗和组穴配伍后的治疗中对Elisa法检测的细胞因子和高效液相色谱法检测的神经递质有不同程度的调节作用。但对PCR法检测的GAT-1的表达有着共同的降低调节作用。从RNA水平证实了蒙医灸疗镇静催眠作用机制。证实了施灸赫依三穴在单穴治疗和组穴配伍治疗中均有镇静催眠作用。.【科学意义】本课题拟用蒙医灸疗取有效腧穴—“顶会穴”、 “赫依穴”、“黑白际穴”(统称赫依三穴)，作用于失眠模型大鼠，观察失眠大鼠行为学变化，运用Elisa、高效液相色谱、PCR方法，检测大鼠脑干组织中HPA轴相关调节因子的表达，探寻赫依三穴组穴配伍规律、并对其治疗作用加以验证，进一步揭示了蒙医灸疗治疗失眠作用机制，为临床病证辩治提供了科学依据。