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研究生:吳怡蓁
研究生(外文):WU, YI-CHEN
論文名稱:添加麩醯胺及魚油之復甦液對創傷出血性休克大鼠發炎反應及標的器官損傷的影響
論文名稱(外文):The effects of glutamine and fish oil-supplemented resuscitation fluids on inflammatory response and target organ injury in rats with trauma and hemorrhagic shock
指導教授:羅慧珍
指導教授(外文):LO, HUI-CHEN
口試委員:張子明李建興
口試委員(外文):CHANG,TZU-MINGLI, CHIEN-HSING
口試日期:2016-12-09
學位類別:碩士
校院名稱:輔仁大學
系所名稱:營養科學系碩士班
學門:醫藥衛生學門
學類:營養學類
論文種類:學術論文
論文出版年:2016
畢業學年度:105
語文別:中文
論文頁數:150
中文關鍵詞:出血性休克復甦液麩醯胺魚油發炎反應氧化壓力
外文關鍵詞:hemorrhagic shockresuscitation fluidglutaminefish oilinflammationoxidation
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出血性休克常發生於創傷、手術及婦女分娩等狀況。患者因血液大量流失,血壓迅速下降、組織器官能量及供氧不足,引發多重器官衰竭(multiple organ failure, MOF)。臨床治療方法為盡速給予患者復甦液,以提升血壓及供氧量,但復甦的同時會促使活性氧分子(reactive oxygen species, ROS)增加,誘導發炎反應及器官損傷。研究指出魚油具有抗發炎的作用,當細胞膜上含較多魚油時,免疫細胞分泌促發炎細胞激素的能力會降低。此外,麩醯胺被證實是免疫細胞及腸道細胞的主要能量來源。因此,本實驗欲探討單獨或同時補充魚油及麩醯胺對出血性休克及復甦(trauma hemorrhagic shock and resuscitation, THR)引發之發炎反應及抗氧化能力的影響。實驗將SD雄性大鼠分為六組,除正常對照組(NC組)及假手術組(SM組)外,其餘THR各組皆進行腹部創傷手術及左頸動脈及右頸靜脈插管,接著THR組由動脈連續抽血10分鐘,使平均動脈壓降為30至35 mmHg以誘導出血性休克。維持低血壓60分鐘後,將抽出的血液由右頸靜脈回注,並於10分鐘內輸入抽出的血液及同體積之乳酸林格氏液(Lactated Ringer's solution, LR)(TH組),或同體積且分別添加L-丙胺-L-麩醯胺(L-alanyl-L-glutamine)(GN組)、魚油或L-丙胺-L-麩醯胺及魚油(GF組)之LR。SM組及THR各組大鼠另行給予連續慢速輸入復甦液42小時後犧牲。故本實驗期能利用補充麩醯胺及魚油之復甦液,可減緩THR所引起的全身及局部器官發炎反應及器官受損現象。結果顯示,本實驗的THR模式會導致大鼠紅血球數量、血紅素、血容積比下降,且會造成血中白蛋白的流失及抗氧化能力下降,此外也發現THR模式會使血漿、肺臟及空腸中的骨隨過氧化化酶(myeloperoxidase, MPO)的活性、肺臟介白素(interleukin, IL)-8含量及腎臟IL-4、IL-8含量降低,而可能會增加體內蛋白質的利用及紅血球的脂質過氧化作用、超氧化物歧化酶(superoxidase dismutase, SOD)及麩胱甘肽過氧化酶(glutathione peroxidase, GPx)活性。THR大鼠給予單獨添加麩醯胺之復甦液,則會降低血漿促發炎細胞激素、肝臟蛋白質氧化作用及紅血球(red blood cell, RBC)脂質過氧化的情形。若給予單獨添加魚油之復甦液,則會降低血漿、肝臟、腎臟促發炎細胞激素及肺臟黏附因子的分泌,而也降低紅血球的脂質過氧化作用。若同時給予THR大鼠添加麩醯胺及魚油之復甦液,則會顯著降低血漿、腎臟促發炎細胞激素的分泌及增加腎臟及空腸的抗發炎作用,也會提升血漿及RBC的抗氧化情形。綜上所述,本實驗的THR模式會降低全身抗氧化作用及增加空腸發炎情形,而給予THR大鼠同時添加麩醯胺及魚油之復甦液則會減緩發炎反應及增加抗氧化作用,而降低THR所造成的全身及局部器官之損傷。
Hemorrhagic shock is an emergent condition that usually caused by trauma, surgery, and parturition. When patients suffer from rapid blood loss, the hypovolemia may result in the decreases in energy and oxygen delivery which cause multiple organ failure (MOF). In clinical practice, patients are administered with resuscitation fluid rapidly to improve blood pressure and oxygen supply. However, resuscitation may elevate the production of reactive oxygen species (ROS) which induce inflammatory response and organ damages. Studies showed that fish oil have the anti-inflammatory activity. When fish oil concentration of the cell membrane is increased, the immune cells have decreased ability to produce pro-inflammatory cytokines. In addition, glutamine has been confirmed to be the main source of energy for immune cells and intestinal cells. Therefore, the aim of the present study was to investigate the effects of single or combination treatment of fish oil- and glutamine-supplemented resuscitation fluids on the THR-induced inflammatory responses and antioxidant activity. Male SD rats were divided into 6 groups, expect the control (NC group) and sham-operated (SM group) groups the other THR groups were suffered with 5 cm midline laparotomy and catheterization in the left carotid artery and right jugular vein. The blood was withdrawn from the left carotid artery to reach a mean arterial pressure 30 to 35 mmHg within 10 min for hemorrhagic shock induction. After 60 minutes of hypovolemia, rats were resuscitated with shed blood and equal volume of lactate Ringer’s solution without (TH group) or with L-alanyl-L-glutamine (GN group), fish oil (FA group) or L-alanyl-L-glutamine plus fish oil (GF group) within 10 minutes and followed by continuouslow infusion rate (~1.4 ml/h) of resuscitation fluids. After 42 h, rats were executed and we expect to demonstrate that glutamine- and fish oil-supplemented resuscitation fluids may attenuate THR-induced inflammatory response and target organ injury in rats. The results show that THR may decreased RBC count, hemoglobin and hematocrit, and the loss of blood albumin and the decrease of antioxidant capacity.Then THR model may lead to decrease plasma, pulmonary and intestinal MPO activity and the pulmonary content of interleukin (IL)-8, the content of IL-4 and IL-8 in the kidney were decreased.The utilization of protein in plasma and lipid peroxidation, SOD and GPx activities in RBC could be increased. THR rats were supplemented with L-alanyl-L-glutamine resuscitation fluid to reduce the plasma pro-inflammatory cytokines, liver protein carbonylation and RBC lipid peroxidation. However, THR rats were supplemented with fish oil resuscitation fluid may decrease the concentration of pro-inflammatory cytokines in plasma, liver, kidney and reduce the pulmonary adhesion factor secretion, but also reduce the lipid peroxidation of RBC. In addition, THR rats were supplemented with L-alanyl-L-glutamine and fish oil resuscitation fluids may significantly reduce the plasma, kidney and pro-inflammatory cytokine secretion and increase the anti-inflammatory in the kidney and jejunum, and may lead to enhance the plasma and RBC antioxidative capacity. In summary, our THR model in this study may reduce the systemic antioxidative capacity and enhance jejunum inflammation. THR rats were supplemented with L-alanyl-L-glutamine and fish oil resuscitation fluids may alleviate inflammatory response and increase the antioxidant effect to reduce the THR-induced inflammatory response and target organ injury.
中文摘要 I
Abstract III
致謝 VI
目錄 VIII
表目錄 XIV
圖目錄 XV
縮寫 XVIII
第一章 1
前言 1
第二章 4
文獻回顧 4
一、 創傷出血性休克及復甦 4
A. 生理變化 4
B. 血液學變化 6
C. 多重性器官衰竭 7
1. 肝臟損傷 7
2. 肺臟損傷 8
3. 腎臟損傷 8
4. 小腸損傷 9
D. 病理機制 9
1. 發炎反應 10
2. 氧化壓力 12
3. 細胞凋亡及壞死 16
二、 創傷出血性休克之復甦治療 16
A. 靜脈注射液-乳酸林格氏液 17
B. 胺基酸輸注液:胺基酸 18
C. 靜脈注射脂肪輸注乳劑-脂肪酸 19
1. 長鏈脂肪酸 19
2. 中鏈脂肪酸 19
3. n-6多元不飽和脂肪酸 19
4. n-3多元不飽和脂肪酸 20
三、 麩醯胺 20
A. 能量來源 24
B. 腸道上皮細胞完整性 24
C. 抗氧化能力 25
D. 調節免疫及發炎反應 25
四、 魚油 27
A. 發炎基因調控 27
B. 免疫調節及細胞激素 28
C. 發炎脂質代謝產物 29
D. 發炎後回復機制 30
第三章 33
目的與假說 33
第四章 35
實驗方法與材料 35
一、 實驗動物 35
A. 創傷出血性休克動物模式 35
B. 實驗設計與分組 36
二、 分析方法 38
A. 血液與生化分析 38
B. 肺臟與空腸水腫情形 39
C. 空腸質量的變化 39
D. 血漿及各組織器官的一氧化氮及細胞激素分析 39
E. 血漿及各組織器官骨髓過氧化酶活性反應 40
F. 血漿及各組織器官之蛋白羰基含量 41
G. 血漿與紅血球脂質過氧化物 42
H. 紅血球酵素型抗氧化分析 43
1. 超氧化物歧化酶酵素活性分析 43
2. 過氧化氫酶酵素活性分析 44
3. 麩胱甘肽過氧化酵素活性分析 44
I. 血漿及紅血球非酵素型抗氧化酵素分析: 45
1. 硫醇基濃度 45
2. 氧化型麩胱甘肽與還原型麩胱甘肽之濃度 46
J. 組織器官之細胞增生 48
K. 組織器官之細胞凋亡 49
三、 統計方法 50
第五章 51
結果 51
一、 創傷出血性休克大鼠復甦後體重、平均動脈壓及復甦液體積 51
二、 全血球計數之變化 52
三、 血液生化值之變化 54
四、 血漿發炎媒介物及氧化狀態 56
A. 血漿發炎媒介物之變化 56
B. 血漿中氧化物及抗氧化物之變化 57
五、 紅血球氧化物質及抗氧化作用之影響 65
六、 各器官組織重量變化及組織型態 72
七、 肺臟與空腸水腫情形 72
八、 肝臟發炎媒介物、細胞凋亡與增生及蛋白質氧化情形 83
A. 肝臟發炎媒介物之變化 83
B. 肝臟細胞凋亡及增生作用 84
C. 肝臟蛋白質氧化情形 85
九、 肺臟發炎媒介物、細胞凋亡與增生及蛋白質氧化情形 91
A. 肺臟發炎媒介物之變化 91
B. 肺臟細胞凋亡及增生作用 92
C. 肺臟蛋白質氧化情形 93
十、 腎臟發炎媒介物、細胞凋亡與增生及蛋白質氧化情形 99
A. 腎臟發炎媒介物之變化 99
B. 腎臟細胞凋亡及增生作用 100
C. 腎臟蛋白質氧化情形 100
十一、空腸質量、發炎媒介物、細胞凋亡與增生及蛋白質氧化情形 106
A. 空腸組織質量變化 106
B. 空腸發炎媒介物之變化 108
C. 空腸細胞凋亡及增生作用 109
D. 空腸蛋白質氧化情形 109
第六章 115
討論 115
一、 THR之生理變化 115
二、 THR發炎反應之影響 115
三、 THR氧化壓力之影響 116
四、 單獨添加麩醯胺復甦液之生理、發炎反應及氧化壓力變化 117
五、 單獨添加魚油復甦液之生理、發炎反應及氧化壓力變化 119
六、 同時添加麩醯胺及魚油復甦液之生理、發炎反應及氧化壓力變化 121
七、 綜論 121
第七章 124
結論 124
第八章 125
參考資料 125
附錄 142


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