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標題: | 在大鼠模式下探討臂旁核中內生性鴉片調控機制與電針刺激促進睡眠之作用 Endogenous opiates in the parabrachial nucleus mediate the electroacupuncture-induced sleep activities in rats |
作者: | Tzu-Wen Wang 王姿雯 |
指導教授: | 張芳嘉(Fang-Chia Chang) |
關鍵字: | 經絡,穴道,電針刺激,臂旁核,孤獨徑核,內生性鴉片系統,迷走神經, meridian,acupoint,electroacupuncture,parabrachial neucleus,nucleus of solitary tract,endogenous opioidergic system,vagus nerve, |
出版年 : | 2014 |
學位: | 碩士 |
摘要: | 針灸是中醫使用已久的治療方式。中醫學根據古中國哲學思想做為基礎論述,形成一套有別於西方醫學之治療系統。黃帝內經被認為是中醫論述之根本,道出萬物與人體皆具有陰陽兩面向並相輔相成。其中對於維持生命最主要物質為”氣”,用以調節陰陽平衡。若”氣”淤塞,陰陽失衡,人體便容易虛弱生病。筋絡則是”氣”所運行之通路,在筋絡特定位置上有”氣”與外接相互接觸點,稱為”穴道”。針灸便是透過針刺刺激穴道,使得阻塞之”氣”得以疏通,使陰陽平衡,回復健康。電針刺激為針灸之改良方式,利用電流透過金屬灸針此機穴位以達成與針灸相同之治療效果。自1970年代即有科學家開始探討針灸治療與電針刺激用以治療疾病之機制,其中以止痛方面最廣為人知。許多中醫師也常利用電針刺激治療失眠,且效果顯著,但是對於電針刺激治療睡眠之機制仍然無法確定。對於電針刺激所調節之機制,科學家們認為主要與自主神經與中樞神經這兩大神經內分泌系統有關,其會活化迷走神經並抑制交感神經,進而改善身體疾病。血清素/單胺性神經系統與內生性鴉片系統為最早被提出電針刺激所活化之機制,膽鹼性系統也被提出與針刺激治療有關。電針刺激安眠穴以治療失眠之路徑為經由迷走神經將訊息傳入孤獨徑核之路徑與活化膽鹼性機制已被證實,本實驗室之前也證實了內生性鴉片系統也為電針刺激安眠穴所誘發的機制之一。在活化了迷走神經之後,訊息便會傳遞入臂旁核中,並調控大腦皮質睡眠。
本實驗將接續之前實驗,證實孤獨徑核之上游轉接核區-臂旁核是否為電針刺激安眠穴所誘發之內生性鴉片系統的調控路徑之一。本實驗分為麻醉組別與無麻醉組別。在麻醉狀況下,麻醉藥zoletil會抑制大鼠前三小時之睡眠,但並不影響電針刺激刺激之效果。電針刺激安眠穴不管大鼠是否在麻醉狀態下,皆能有效促進大鼠在暗期之睡眠。在麻醉底下,分別微量注射四種opioid receptor antagonists進入臂旁核中,皆能有效抑制電針刺激所增加之非快速動眼睡眠;另一方面,在非麻醉狀態下進行電針刺激,μ-與δ-opioid receptor antagonist (naloxonazine與naltrindole)並沒辦法有效抑制電針刺激所誘發之睡眠,反而κ-opioid receptor antagonist具有最有效之作用。由此可知,在臂旁核中,電針刺激安眠穴最主要利用κ-opioid receptor來傳遞訊息。由電針刺激不含有穴位之腋窩則是證實了電針刺激位置必須在特定作用之穴位上,在任何一不是穴位之位置刺激並不會獲得理想之治療效果。最後,也證實了微量注射四種opioid receptor antagonists入臂旁核中並不會影響大鼠本身之睡眠活動。 Previous studies indicate that electroacupuncture (EAc) of Anmian acupoint (EX 17) enhances vagal activity and changes the synaptic morphology in the nucleus tractus solitarius (NTS) via vagus nerve by different EAc stimulus frequencies. The goal of this study is to investigate the role of parabrachial nucleus (PBN), which receives afferents from NTS, in the EAc-induced sleep alterations. Total of sixty-nine male Sprague-Dawley rats, weighted 270-300 g, was used. We microinjected four types of opioid receptor antagonists: naloxone (broad-spectrum opioid receptor antagonist), naloxonazine (μ-opioid receptor antagonist), naltrindle (δ-opioid receptor antagonist), nor-binaltorphimine (κ-opioid receptor antagonist), into the PBN before the EAc stimulation in dosage of 10 μg/0.5 μl. The total volume used for each microinjection was 0.5 μl. A 30-minute EAc stimulation was performed at the beginning of the dark period in a 12:12h light:dark cycle. The frequency of EAc used in this experiment was 10 Hz, and the EEGs were recorded after EAc stimulation and lasted for 24 hours. Three groups of experiments were designed to confirm this study, including EAc with and without anesthesia group, blocker only group. Un-paired Student T test was used for statistical analysis. Our preliminary results indicated that EAc significantly enhanced non-rapid eye movement (NREM) sleep after the 30-minute EAc stimuli in the EAc with anesthesia group. Microinjection of naloxone, naloxonazine, naltrindole, nor-binaltorphimine into the PBN 30 minutes prior to the EAc stimulation reduced the percentage of NREM sleep. μ- and δ-opioid receptor antagonists hadn’t well function to block the EAc-induced enhancement of NREM sleep in the EAc without anesthesia group. In contrast, κ-opioid receptor did the efficiency work to deliver EAc message. It had another evidence to prove that opioid receptor antagonists were not influencing the normal sleep of rats. Our results indicated that 10 Hz EAc stimulation of Anmian acupoints increased spontaneous NREM sleep during the dark period, suggesting the sleep improvement for EAc stimuli of anmian acupoints. Microinjection of four opioid receptor antagonists directly into the PBN blocked EAc-induced enhancement of NREM sleep, indicating the role of endogenous opioids in the PBN. This observation further demonstrated endogenous opioids in the PBN, the relay nucleus of NTS, mediate EAc-induced sleep alterations. |
URI: | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/57897 |
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