記錄
編號
12114
狀態
G0498866159
助教
查核
建檔完成
索書
號
查核完成
學校
名稱
輔仁大學
系所
名稱
營養科學系
舊系
所名
稱
學號
498866159
研究
邱郁雯
生(中)
研究
Chiu, Yu -Wen
生(英)
論文
名稱
(中)
論文
名稱
(英)
口服麩醯胺酸、瓜胺酸及抗氧化維生素改善小腸缺血再灌流大鼠之腸道
損傷及發炎反應
Oral administration of glutamine, citrulline and antioxidative vitamins improve
intestinal damage and inflammatory response in rats with intestinal ischemia and
reperfusion
其他
題名
指導
教授
(中)
羅慧珍
指導
教授
(英)
Lo, Hui-Chen
校內
全文
開放
日期
2017.2.21
校外
全文
2017.2.21
開放
日期
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不開
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由
電子
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送交
國圖.
國圖
全文 2017.2.21
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01
學位
類別
碩士
畢業
學年
度
100
出版
年
101
語文
別
英文
關鍵
小腸缺血再灌流 麩醯胺酸 瓜胺酸 維生素 C 維生素 E 小腸損傷 發炎反應
字(中)
關鍵 intestinal ischemia-reperfusion glutamine citrulline vitamin C vitamin E intestinal
字(英) damage inflammatory response
摘要
(中)
小腸缺血再灌流(ischemia and reperfusion, I/R)損傷常發生於急性腸繫膜缺
血、腸阻塞、器官移植、大量失血及敗血性休克患者。研究指出，小腸
I/R 引發之腸黏膜局部損傷易導致全身性發炎及多重器官衰竭，甚至造成
死亡。我們先前的研究證實，口服麩醯胺酸(Gln)及瓜胺酸(Cit)可顯著改
善小腸 I/R 大鼠的腸道與全身性發炎反應。亦有研究顯示，小腸 I/R 前給
予抗氧化劑，如維生素(vitamin) C 及 E，可降低脂質過氧化反應及氧化壓
力，進而減緩小腸損傷。故本研究進一步探討，合併補充 Gln、Cit 及
vitamin C、E 對小腸 I/R 誘導腸道損傷及發炎反應之改善，是否具有加成
作用。雄性 Wistar 大鼠分為正常對照組(NC 組)及施予上腸繫膜動脈阻塞
手術，誘導小腸缺血 60 分鐘後再予以灌流之實驗組。大鼠於灌流前 15
分鐘及灌流後 3、9 及 21 小時，分別灌餵安慰劑(IR 組)、Gln 與 Cit (AA
組)、vitamin C 與 E (CE 組)、或 Gln 與 Cit 合併 vitamin C 與 E (AV 組)，
並於灌流後 24 小時犧牲。結果顯示，IR 組血清白蛋白與葡萄糖濃度及
血漿精胺酸(Arg)、腫瘤壞死因子(TNF)-r與干擾素(IFN)-r含量顯著低於
NC 組，且小腸絨毛長度與 DNA 含量及小腸 Gln、脯胺酸、鳥胺酸與蛋
白質氧化物含量顯著低於 NC 組。此外，IR 組血液白血球數目、血清肝
臟與腎臟功能指標及血漿間白素(IL)-6 與ㄧ氧化氮代謝物(NOx)濃度顯著
高於 NC 組，且小腸 IL-2、IL-6、TNF-r、IFN-r與脂質過氧化物
(TBARS)含量、小腸骨髓過氧化酶(MPO)活性及小腸細胞凋亡顯著高於
NC 組。AA、CE 及 AV 組血清白蛋白顯著高於 IR 組，且血漿 IL-6 及小
腸 TBARS 含量顯著低於 IR 組。AA 及 AV 組血漿 Arg 及 Cit 濃度及小腸
Cit 含量顯著增加；CE 及 AV 組肝功能指標及小腸 MPO 活性顯著降低，
且血漿與小腸 vitamin E 含量及小腸 vitamin C 含量顯著增加。此外，AA
組小腸絨毛長度顯著增加，且血漿 NOx 濃度顯著降低；CE 組血漿 NOx
濃度顯著降低；AV 組小腸肌肉層 DNA 和蛋白質含量顯著增加，且血漿
TNF-r、IFN-r、TBARS 濃度、小腸 TNF-r和 IL-6 含量及小腸細胞凋
亡顯著降低。二因子變異數分析之結果顯示，小腸 I/R 大鼠合併補充
Gln、Cit、vitamin C 及 E，於血漿與小腸 vitamin E 含量的增加及血漿
TNF-r與 IFN-r濃度與腸道細胞凋亡現象的降低具有加成作用。本實驗
結果建議，口服麩醯胺酸、瓜胺酸及抗氧化劑可顯著改善小腸缺血再灌
流大鼠之腸道損傷及發炎反應，且合併使用具有減緩腸道損傷及全身性
發炎反應之加成作用。
摘要
(英)
Intestinal ischemia and reperfusion (I/R)-induced damage is a common event in
patients with acute mesenteric ischemia, intestinal obstruction, organ
transplantation, and hemorrhagic and septic shock. Several studies indicated that
intestinal I/R-induced intestinal mucosa damage may cause systemic inflammation
and multiple organ failure and further lead to death. Our previous study
demonstrated that oral administration of glutamine (Gln) and citrulline (Cit)
significantly improved the intestinal and systemic inflammation in rat with
intestinal I/R. Recently, it has been revealed that pre-administration of
antioxidants, such as vitamin C and E, may attenuate the intestinal I/R-induced
damage by reducing lipid peroxidation and oxidative stress and further alleviate
the intestinal damage. Therefore, the aim of this study was to further investigate
that whether the combination treatment of oral Gln, Cit, vitamin C and E has an
additive effect on improving intestinal I/R-induced intestinal damage and
inflammatory response. Male Wistar rats were divided into a normal control (NC)
group and experimental groups suffered with occlusion of superior mesenteric
artery for 60 min to induce intestinal ischemia and with subsequent reperfusion.
Fifteen minutes before and 3, 9 and 21 hours after reperfusion, rats with intestinal
I/R were gavaged with placebo (IR group), Gln plus Cit (AA group), vitamin C
plus E (CE group), or combination of Gln, Cit, vitamin C and E (AV group). Rats
were executed 24 hours after reperfusion. Our results showed that the IR group
had significantly decreased serum albumin and glucose concentrations in the
serum and arginine, tumor necrosis factor (TNF)-rrand interferon (IFN)-r
concentrations in the plasma and significantly decreased villus height, DNA
content, Gln, proline, ornithine and carbonyl protein contents in the jejunum
compared to the NC group. In addition, the IR group had significantly increased
circulating numbers of white blood cells, biomarkers of hepatic and renal function,
plasma interleukin (IL)-6 and nitrate/nitrite (NOx), intestinal IL-2, IL-6, TNF-r
and IFN-r content, intestinal myeloperoxidase (MPO) activity and thiobarbituric
acid reactive substances (TBARS) content and apoptosis compared to the NC
group. The AA, CE and AV groups had significantly increased serum albumin and
decreased plasma IL-6 and intestinal TBARS contents compared to the IR group.
The AA and AV groups have significantly increased plasma arginine and citrulline
concentrations and intestinal citrulline content; the CE and AV groups have
significantly decreased biomarkers of liver function and intestinal MPO activity
and increased plasma and jejunal vitamin E content and intestinal vitamin C
content compared to the IR group. Moreover, the AA group had significantly
increased intestinal villus height and significantly decreased plasma NOx; the CE
group had significantly decreased plasma NOx; and the AV group had
significantly increased intestinal muscularis DNA and protein contents and
decreased plasma TNF-r, IFN-r and TBARS levels, intestinal TNF-r and IL6 content and enterocytic apoptosis compared to the IR group. The results of twoway ANOVA analysis indicated that combination treatment of glutamine,
citrulline, vitamin C and E had additive effects on increasing plasma and intestinal
vitamin E levels and on decreasing plasma TNF-r and IFN-r concentrations
and enterocytic apoptosis in intestinal I/R rats. The results of this study suggest
that oral administration of glutamine, citrulline and antioxidants may significantly
improve intestinal damage and inflammatory response in intestinal I/R rats. In
addition, combination treatment may have additive effects on attenuating intestinal
I/R-induced intestinal damage and systemic inflammatory response.
論文
目次
ABSTRACT IN CHINESE i ABSTRACT IN ENGLISH iii
ACKNOWLEDGEMENTS vi LIST OF TABLES xii LIST OF FIGURES xiii
ABBREVIATION xv CHAPTER 1 INTRODUCTION 1 CHAPTER 2
LITERATURE REVIEW 3 I. Intestinal ischemia and reperfusion (I/R) 3 A.
Clinical relevance 3 B. Intestinal ischemia and reperfusion stress 4 1. Intestinal
mesenteric blood flow 4 2. Ischemia and reperfusion 9 3. Reactive oxygen species
(ROS) 10 4. Polymorphonuclear neutrophils (PMNs) 13 5. Local damages and
inflammatory response 14 a. Proinflammatory cytokines 14 b. Nitric oxide (NO)
15 c. Intestinal mass and morphology 17 d. Enterocyte apoptosis 20 e. Crypt cell
proliferation 21 6. Remote organ failure 22 a. Lung injury 23 b. Liver injury 24 c.
Kidney injury 25 7. Metabolic responses 25 8. Therapeutics 27 a. Surgery 27 b.
Pharmacological treatments 28 c. Alternative treatments 28 II. Amino acid
supplementation 29 A. Amino acid interorgan metabolism 29 B. Glutamine 33 1.
The role of physiology 33 2. Therapeutic implications 35 C. Citrulline 36 1. The
role of physiology 36 2. Therapeutic implications 37 III. Antioxidant
supplementation 38 A. Antioxidant system in I/R 38 B. Ascorbic acid 39 1. The
role of physiology 39 2. Therapeutic implications 42 C. r-Tocopherol 42 1. The
role of physiology 42 2. Therapeutic implications 43 CHAPTER 3 AIMS AND
HYPOTHESIS 45 CHAPTER 4 MATERIALS AND METHODS 47 I. Animals
47 II. Experimental design 47 III. Analytic measurements 52 A. Hematological
and serological analysis 52 B. Amino acid levels in the plasma and jejunum 52 C.
Ascorbic acid levels in the plasma and jejunum 54 D. r-Tocopherol levels in the
plasma and jejunum 55 E. Nitrate/nitrite concentration in the plasma and jejunum
56 F. Cytokine concentrations in the plasma and jejunum 57 G. Jejunal
morphology 57 H. Jejunal mass 58 I. Jejunal apoptotic index 58 J. Jejunal
myeloperoxidase (MPO) activity 59 K. Thiobarbituric acid reactive substances
(TBARS) levels in the plasma and jejunum 60 L. Carbonyl protein (CP) content in
the jejunum 61 IV. Statistical analysis 62 CHAPTER 5 RESULTS 63 I. Body
weight and weights of organs and tissues 63 II. Blood and serum substrate
concentrations 63 III. Amino acid levels in the plasma and jejunum 64 IV.
Ascorbic acid levels in the plasma and jejunum 66 V. r-Tocopherol levels in the
plasma and jejunum 66 VI. The architecture and composition of the jejunum 67
VII. Jejunal villus apoptosis 68 VIII. Jejunal myeloperoxidase (MPO) activity 68
IX. Nitrate/nitrite and cytokine concentrations in the plasma and jejunum 69 X.
Thiobarbituric acid reactive substances (TBARS) levels in the plasma and jejunum
70 XI. Carbonyl protein (CP) content in the jejunum 71 CHAPTER 6
DISCUSSION 102 CHAPTER 7 CONCLUSION 112 CHAPTER 8
REFERENCES 114 APPENDICES 135 APPENDIX I. Oral presentation in
Nutrition Society of Taiwan 2011 135 APPENDIX II. Poster presentation in
Health Food Society of Taiwan 2010 139
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