臺灣博碩士論文加值系統

血清素在酸棗仁湯之睡眠調控機制中所扮演的角色
(The involvement of sertonin in Suanzaorentang-induced sleep regulation)

研究生 林峻邦

指導教授 張芳嘉博士

中國醫藥大學 中國醫學研究所


酸棗仁湯是臨床上中醫常應用在睡眠障礙的方劑，首載於東漢時期張仲景所著的金匱要略中，主治為虛勞虛煩不得眠。臨床上用於治療因肝陰不足，心血虧虛，肝虛血燥而導致的虛性亢奮，療效顯著。本研究主要探討中國醫學傳統方劑酸棗仁湯對睡眠調控中血清素(serotonin)所扮演的角色。在大鼠腦部手術植入記錄電極及側腦室注射導管，利用腦波圖(electroencephalogram，EEG)及locomotor activity來觀察酸棗仁湯是否影響大鼠睡眠。並透過餵食大鼠酸棗仁湯後，再以血清素(serotonin)的拮抗劑，包括5-HT1A之antagonist NAN-190、5-HT2之antagonist ketanserin及5-HT3之antagonist打入大鼠的側腦室(lateral ventricle)內來觀察大鼠腦波圖(EEG)之變化，來探討酸棗仁湯影響大鼠睡眠的機轉是否透過血清素這個路徑。並從酸棗仁湯中依序抽取出川芎、茯苓、知母、甘草、酸棗仁一味藥物，形成酸棗仁湯Ⅱ、Ⅲ、Ⅳ、Ⅴ、Ⅵ五個方劑，觀察這五個方劑是否和原方劑有一樣的效果，試著找出酸棗仁湯中主要影響大鼠睡眠的藥物。

本研究結果顯示：
酸棗仁湯4 g/kg組明顯增加大鼠NREMS(非快速動眼期睡眠; non-
rapid-eye-movement sleep)的量達33.02 ± 2.35 %，比對照組25.12 ± 2.15 % 多將近8 % 的量(p < 0.05)。而酸棗仁湯0.5 g/kg, 1 g/kg, 2.0 g/kg組，其NREMS的量，無法和對照組有明顯的區分。至於REMS (快速動眼期睡眠; rapid-eye-movement sleep)方面，並無明顯增加，無統計學上的意義。NAN-190 (5-HT1A antagonist) , ketanserin (5-HT2 antagonist) 及5-HT3之antagonist 皆能使餵食酸棗仁湯大鼠的NREMS的量降低(p < 0.005)。但三個5-HT的拮抗劑之間並無明顯的區別。在REMS方面，並無統計學上的意義。

酸棗仁湯抽取任任一藥味後，五個方劑的大鼠NREMS及REMS並無明顯變化，統計學上無明顯差異。

不同劑量 (1 nmole, 5 nmole, 10 nmole)的ketanserin及NAN-190投予大鼠後，皆可以使餵食酸棗仁湯4 g/kg的大鼠之NREMS的量降低(p < 0.05)，而不同劑量間所降低的差異並不明顯，在統計學上並無明顯差異。而REMS量則無明顯降低。

綜合以上結果，酸棗仁湯有增加大鼠NREMS的量，可推測酸棗仁湯有促進大鼠睡眠的作用。血清素的拮抗劑可以使餵食酸棗仁湯大鼠所增加的NREMS量降低，由此可以推測酸棗仁湯所增加大鼠NREMS的量，是經由血清素所媒介。抽掉酸棗仁湯中任一味藥物後，大鼠的NREMS量並無明顯改變，並未如預期可以找到酸棗仁湯中主要作用的藥物。但是酸棗仁單味藥，在藥理學研究上是可以促進睡眠。酸棗仁湯五種藥物的組合促使睡眠增加，和中醫方劑學中的君、臣、佐、使配伍理論有密切相關，各藥物之間因配伍不同而產生不同的作用，但酸棗仁湯五種藥物因互相調節而使其發揮促進睡眠的作用是確定的。

關鍵詞：酸棗仁湯、血清素、睡眠

The involvement of serotonin in Suanzaorentang-induced sleep regulation

Chun-Pang Lin

Major professor：Fang-Chia Chang, Ph.D.

Institute of Chinese Medical Science, China Medical University, Taiwan, ROC

ABSTRACT

Suanzaorentang (SZR Tang) is a compound of traditional Chinese medicine, which is widely used for treatment in insomnia. It was first demonstrated in the “Jin Kui Yao Lve”written by Chong-Jing Chang. In this study, we investigated the hypnotic effect of SZR Tang in rats. Furthermore, we examined the role of serotonin (5-hydroxytryptamine, 5-HT) in SZR Tang-induced sleep alterations by microinjection of 5-HT receptor antagonists into the lateral ventricle of the rat. We also drew out anyone of the ingredients from SZR Tang to determine whether withdrawing anyone ingredient from Suanzaorentang affects its hypnotic action.

Our results indicate that the high dose (4.0 g/kg) of SZR Tang increased the NREM sleep to from 16.79±1.87 % obtained after baseline to 33.02±2.35 %. However, starch placebo (4.0 g/kg) also increased NREMS to 25.12±2.15 % . REM sleep has no change apparently after both placebo and SZR Tang. All three subtypes of 5-HT antagonists decreased SZR Tang-induced increase in NREM sleep. In addition, our results also demonstrate that no statistical change in both NREM sleep and REM sleep comparing with those from baseline recording when one ingredient was withdrew.

These results suggest that the SZR Tang exhibits the hypnotic effect in rats. The SZR Tang-induced increase in NREM sleep was block by the 5-HT antagonists, suggesting the involvement of serotonin in this mechanism.
When anyone of the ingredient withdraws from SRZ Tang, it exhibits no change in both NREM sleep and REM sleep. This indicates that the total five ingredients of SZR Tang possess the hypnotic effect rather than the prescription lacking of anyone ingredient.

Keywords：Suanzaorentang, serotonin, sleep.

目錄

頁碼

第一章 前言 1
第二章 文獻探討 4
第三章 材料與方法 12
第四章 結果 18
第五章 討論 41
第六章 結論 45
參考文獻 46
英文摘要 52
謝辭 54

圖目錄
頁碼
圖3.1：紀錄電極(·)及anchor (o)植入位置。 14

圖4.1：酸棗仁湯, placebo, baseline在dark-period 中 NREMS的變化 19

圖4.2：酸棗仁湯, placebo, baseline在dark-period 中REMS的變化 20

圖4.3：酸棗仁湯不同劑量(0.5g, 1, 2, 4 g/kg), placebo, baseline在 21
dark-period中NREMS量的比較。

圖4.4：酸棗仁湯不同劑量(0.5 g, 1, 2, 4 g/kg), placebo, baseline在 22
dark-period中REMS量之比較

圖4.5：注射NAN-190, ketanserin, anti-5-HT3 (10 nmole)對餵食 24
大鼠SZR Tang (4 g/kg) NREMS量的變化。

圖4.6：注射NAN-190, ketanserin, anti-5-HT3 (10 nmole) 對餵食 25
大鼠SZR Tang (4 g/kg) REMS量的變化

圖4.7：注射NAN-190, ketanserin, anti-5-HT3 (10 nmole) 對餵食 26
大鼠SZR Tang (4 g/kg) NREMS量的比較

圖4.8：注射NAN-190, ketanserin, anti-5-HT3 (10 nmole) 對餵食 27
大鼠SZR Tang (4 g/kg) REMS量的比較

圖4.9：酸棗仁湯Ⅱ, Ⅲ, Ⅳ, Ⅴ, Ⅵ與baseline, placebo在 31
dark-period中NREMS量的比較

圖4.10：酸棗仁湯Ⅱ, Ⅲ, Ⅳ, Ⅴ, Ⅵ與baseline, placebo在 32
dark-period中REMS量的比較

圖4.11：酸棗仁湯4 g/kg 和不同劑量ketanserin (1,5,10nmole)在 35
dark-period中NREMS的變化

圖4.12：酸棗仁湯4 g/kg 和不同劑量NAN-190 (1,5,10nmole) 在 36
dark-period 中NREMS的變化

圖4.13：酸棗仁湯4 g/kg和不同劑量ketanserin (1,5,10 nmole)在 37
dark-period中REMS的變化

圖4.14：酸棗仁湯4 g/kg和不同劑量NAN-190 (1,5,10 nmole)在 38
dark-period中REMS的變化

圖4.15：酸棗仁湯4 g/kg和不同劑量NAN-190 (1,5,10 nmole)在 39
dark-period中NREMS量的比較

圖4.16：酸棗仁湯4 g/kg和不同劑量ketanserin (1,5,10 nmole)在 40
dark-period中NREMS量的比較

表目錄

頁碼

表4.1：不同劑量酸棗仁湯(0.5, 1.0, 2.0, 4.0 g/kg), baseline, placebo 18
在dark-period中NREMS及REMS的量

表4.2：餵食酸棗仁湯(SZR Tang 4 g/kg)及注射NAN-190, ketanserin 23
, anti-5-HT3 對大鼠NREMS及REMS的量

表4.3：酸棗仁湯Ⅱ, baseline, placebo 在NREMS及REMS 28
量的比較

表4.4：酸棗仁湯Ⅲ, baseline, placebo 在NREMS及REMS 29
量的比較

表4.5：酸棗仁湯Ⅳ, baseline, placebo 在NREMS及REMS 29
量的比較

表4.6：酸棗仁湯Ⅴ, baseline, placebo 在NREMS及REMS 30
量的比較

表4.7：酸棗仁湯Ⅵ, baseline, placebo 在NREMS及REMS 30
量的比較

表4.8：酸棗仁湯4 g/kg與餵食酸棗仁湯後注射不同ketanserin劑量 33
(1, 5, 10 nmole) 之NREMS及REMS量之比較

表4.9：酸棗仁湯4 g/kg與餵食酸棗仁湯後注射不同NAN-190劑量 34
(1, 5, 10 nmole) 之NREMS及REMS量之比較

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