睡眠剝奪病理機轉

第二章文獻探討

第二節睡眠剝奪病理機轉

一個人每天所需的睡眠可能是由基因決定的平均8 小時，如果減少他

的睡眠時間會降低白天的警覺性。警覺度降低或嗜睡情形增加，所顯示出來的狀況是注意力不集中、記憶力減退、以及交通事故的危險增加。

許多臨床及動物實驗研究顯示﹕睡眠剝奪會造成免疫力、內分泌、認知、記憶‥等不良之影響(Nisapel N. Cell Mol Life Sci 2007)，也會影響傷口癒合(Landis et al. Research in Nursing and Health. 1997)，甚至與發炎反應有關，換言之，睡眠剝奪會引起發炎反應(Malik SW. et al. Prim Care Clin Office Pract 2005)。

第三章研究方法第一節研究材料

公的大白鼠(Sprague-Dawley rats )8 隻，年齡約 5-6 週大，體重約 300-400 公克。每隻大白鼠腦部皆植入腦波監視器，植入腦波監視器後休息 2 天使傷口癒合，。大白鼠飼養在長60 (公分) × 寬 20(公分)×高60 (公分) 的籠子，且鄰近有同樣飼養的大白鼠陪伴。

雷赫特夏芬(Rechtschaffen)的大白鼠睡眠剝奪實驗設計(圖 5 及 6) ，不管

是哪一組皆要在轉盤上適應5 天。

當實驗組的老鼠想睡時，其腦波的睡眠波形會啟動動物籠的轉動。控制組

的老鼠則可在不轉動的地板自由的睡覺。睡眠剝奪時間為 24 小時。3. 形

態測定方法

利用電腦色特殊軟體計算血管新生內膜及中膜的表面面積比率(圖 7)。

第三節統計方法

新生內膜及中膜的表面面積比率以平均值及標準誤表示，並以 ANOVA 變異 數分析各組之關係。p<0.05 表示有統計學上之差異。

第四章研究結果

第一節描述性統計分析

C 組有兩隻大白鼠，A 組有兩隻大白鼠，B 組有兩隻大白鼠，AB 組有兩隻大白鼠。

1. 每組大白鼠其施行氣球血管形成術後之右側頸動脈血管皆有新生內膜之

現象，而左側頸動脈血管則無新生內膜之現象(圖8 及 9)。

2.血管新生內膜及中膜的表面面積比率﹕C 組為 0.900±0.010，A 組為 1.195±0.125，B 組為 1.435±0.045，AB 組為 1.725±0.021 (表 1)。

第二節推論性統計分析

A 組、B 組、 AB 組的血管內膜及中膜的表面面積比率皆比 C 組有顯著增加，分別各增加了 31.87%, 58.24%, 及 87.91%。且 A 組、B 組的血管新生

內膜及中膜的表面面積比率皆比C 組有顯著低(圖 10)。由此表示睡眠剝奪

會增加老鼠頸動脈損傷後新生內膜增生。

生更明顯。於是我們下一個結論: 睡眠剝奪會使促使氣球血管形成術後之內膜增生。

第二節其他相關性討論

臨床上遇到之狀況:病患預施行冠狀動脈氣球擴張術(PTCA)來治療冠狀動脈疾病，但可能因緊張焦慮而失眠，或施行之後睡不好‥等。所以可以進一步收集臨床上有睡眠障礙或有睡眠呼吸中止症之病患且曾施行冠狀動脈氣球擴張術者，回溯性分析並探討人類睡眠障礙會對冠狀動脈氣球擴張術之後的影響。探討是否發生再狹窄之機會增加? 或血管平滑肌受損更嚴重? 如何預防?

第三節研究限制

1.此研究所用的大白鼠數目較少，每組只有 2 隻，最好每組可以 6 隻以上。

2. 此研究並無測量大白鼠之賀爾蒙或細胞激素，只做組織切片之染色，或許下個論文可以設計其他實驗來探討賀爾蒙或細胞激素之變化。

3.我們只從形態測定方法計算血管新生內膜及中膜的表面面積比率，分析及評估其比率是否會因睡眠剝奪而增加，並無從病理切片染色或免疫生化染色來觀察新生內膜之細胞變化。

4. 此研究大白鼠皆為公的，不知母的大白鼠是否也有同樣的結果，還是會因懷孕或女性賀爾蒙改變其結果。

第六章結論與建議第一節結論

氣球血管形成術前及後之睡眠剝奪增加老鼠頸動脈損傷後新生內皮增生。

第二節建議

1.研究繼續進行，最好累積每組大白鼠數目達 6 隻以上。

2.將來研究之方向:確定睡眠剝奪會造成氣球擴張後之血管增生及探討促進因子。可從分子細胞、病理切片染色或免疫生化染色、蛋白質或發炎細胞激素層次來設計更進一步之實驗深入探討其原因及機轉，或利用組織螢光染色技術來研究局部血管附近之變化。也可加驗大白鼠之細胞激素，如 IL-6、IFN- 、TNF-α‥等。

3.經由本研究，我們建議在臨床上收集有睡眠障礙或有睡眠呼吸中止症之病患且曾施行冠狀動脈氣球擴張術者，回溯性分析其發生冠狀動脈再狹窄之機會是否比一般正常健康者高。

4. 在施行冠狀動脈氣球擴張術之前及後應給予適當之安眠劑或鎮定劑，以免將來增加冠狀動脈再狹窄之機會。

圖

圖 1、血管受損後造成新生內膜增生之機轉模式圖。

血管受損後活化血小板釋放分裂素，使平滑肌肉細胞分裂增生且移行至

內膜，造成血管新生內膜增生。

圖 3、頸動脈氣球血管形成術卡通圖示。

使用 2F-Fogarty 氣球導管，打氣至 1.3kg/cm²，通過大白鼠右側之頸動脈血管 3 次。

圖 4、血管受傷後造成內膜增生之機轉變化圖示﹕新生內膜之增生在 7-14

天達最明顯之程度。

圖 5、雷赫特夏芬(Rechtschaffen)的大白鼠睡眠剝奪實驗設計卡通圖示﹕

右邊為控制組之大白鼠，左邊為實驗組之大白鼠。

圖 6、本實驗之大白鼠睡眠剝奪實驗圖:

右邊為控制組之大白鼠，左邊為實驗組之大白鼠。

EEG = 連線至腦波監視器。

EEG

圖 7、形態測定分析方法:

左邊為 H&E stain，右邊為電腦繪圖指示:紅色部份為新生內膜及黃色部份為中膜。

圖 8、各組左側與右側頸動脈血管橫切圖:

上圖為左側頸動脈血管橫切圖片(a 是大白鼠 C1, c 是大白鼠 B1, e 是大白鼠 B1, d 是大白鼠 AB1,)，下圖為右側頸動脈血管橫切圖片(b 是大白鼠 C1, d 是大白鼠 B1, f 是大白鼠 B1, g 是大白鼠 AB1,)。(H&E stain，放大倍率為 100 倍)

圖 9、頸動脈血管橫切圖片:

a 圖為左側頸動脈血管橫切圖片， b 圖為右側頸動脈血管橫切圖片。(A = adventitia, M= media, N= neointima)

(H&E stain，放大倍率為 400 倍)

GroupC Group A Group B Group AB

Neo/Media

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附錄

附錄一英文部份 SUMMARY

Coronary artery disease can be treated with percutaneous transluminal coronary angioplasty (PTCA). Although this procedure is efficacious in opening stenotic arteries it may also cause injury and inflammation to the vessel producing neointima formation and subsequently restenosis. Sleep deprivation has major effects on early inflammatory response and may produce numerous untoward effects on cardiovascular disease and wound healing. However, whether sleep deprivation may affect injury-induced neointimal proliferation of the vessel is unknown. This preliminary study is to investigate if sleep deprivation will augment balloon angioplasty induced neointimal proliferation in carotid arteries of rats. Rats were randomly assigned to the following four groups: Group C (control group): balloon angioplasty without sleep deprivation. Group A:

balloon angioplasty after 24-hours sleep deprivation. Group B: balloon angioplasty before 24-hours sleep deprivation. Group AB: 24-hours sleep deprivation before and after balloon angioplasty. Twenty four hours sleep

deprivation was performed by the disc-on-water method for the rats in Group A, Group B and Group AB. Balloon injury was performed with all rats anesthetized and afterwards subjected to an injury of the right carotid artery with a

2F-Fogarty balloon catheter. The untouched left carotid artery was used as another (self) control. Thirteen days after the balloon injury, all of the rats were

sacrificed and both carotid arteries were removed. The cross sections were later stained with hematoxylin and eosin (H&E) for morphometric analysis. There were 2 rats in each group. The post injury neointima-to-media area ratio in Group C, Group A, Group B and Group AB were 0.90±0.01, 1.2±0.13,

1.44±0.05 and 1.73±0.02, respectively. There were 31.87%, 58.24%, and 87.91 increase in post-injury neointima-to-media area ratio in Group A, Group B and Group AB, respectively, compared with Group C (p<0.05). In addition, the post injury neointima-to-media area ratios in Group A and Group B were lower than that in Group AB (p<0.05). There were no neointimal proliferations in the left carotid artery for all groups. This preliminary study shows that neointimal proliferation induced by balloon angioplasty is significantly increased whether the 24-hours sleep deprivation was before and/or after balloon angioplasty of rats. In addition, sleep deprivation before and after balloon angioplasty had significantly more neointimal proliferation than sleep deprivation before or after angioplasty alone. We concluded that sleep deprivation significantly augments post-injury neointimal proliferation in carotid artery angioplasty of rats.

KEYWORDS: sleep deprivation, balloon angioplasty, neointimal proliferation

INTRODUCTION

Coronary artery disease develops when a combination of fatty material, calcium and plaque builds up in the arteries that supply the heart with blood and is one of the major causes of death and disability. The disease can be treated with percutaneous transluminal coronary artery angioplasty (PTCA) by passing a Fogarty balloon catheter into the lumen of the narrowing artery.

However, this procedure can cause the medial smooth muscle cells (SMCs)

proliferation within two days and SMCs migration into the intima after four days, where they continue to proliferate for up to two weeks (Olson et al., 2000).

These neointimal proliferations and geometric arterial remodeling subsequently result in restenosis, which develops in 30-50% of angioplasty receivers within six months (Stigwart, 1999; Kimura T et al., 1997; Faxon et al., 1984).

Many candidate molecules that regulate neointimal proliferation following vessel injury have been studied including the rennin-angiotensin system, catecholamines, endothelin-1 (ET-1), natriuretic peptides, thrombin, platelet-derived growth factor (PDGF), transforming growth factor-ß (TGF-ß), fibroblast growth factor, nitric oxide and the estrogen receptor (Berk, 2001;

Pawlowski et al., 1997; Gong et al., 1996; Konneh et al., 1995; Rudic et al., 1998; Iafrati et al., 1997). A numbers of studies carried out in the last decade

在文檔中氣球血管形成術前及後之睡眠剝奪增加老鼠頸動脈損傷後新生內膜增生-初步研究; Sleep Deprivation Before and After Balloon Angioplasty Significantly Augments Post-injury Neointimal Proliferation in Carotid Arteries of Rats – Preliminary Study (頁 17-79)

第二章 文獻探討

第二節 睡眠剝奪病理機轉

第二章文獻探討

第二節睡眠剝奪病理機轉