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研究生:王琦嫻
研究生(外文):Chi-Hsien Wang
論文名稱:以介白素6與介白素10評估敗血症大鼠之預後與白虎湯之治療效應
論文名稱(外文):The Evaluation of Outcome Prediction and Pai-hu-tang Effect on Septic Rats by IL6 and IL10
指導教授:蘇奕彰蘇奕彰引用關係
學位類別:博士
校院名稱:中國醫藥大學
系所名稱:中國醫學研究所博士班
學門:醫藥衛生學門
學類:醫學學類
論文種類:學術論文
論文出版年:2006
畢業學年度:94
語文別:中文
論文頁數:83
中文關鍵詞:敗血症盲腸結紮與穿孔手術線性鑑別方程式
外文關鍵詞:sepsiscecal ligation and puncturelinear discriminant function
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敗血症的高死亡率一直是臨床上的難題,由於其病程發展快速,病理機轉複雜,在治療上實有其難以掌握之處。敗血症發生後,無法控制的細胞激素風暴是病人死亡的重要原因,因此,發炎細胞激素與抗發炎細胞激素的變化是病情是否惡化的重要關鍵。爲了瞭解發炎細胞激素以及抗發炎細胞激素與死亡的相關性,本研究選擇介白素6與介白素10分別作為發炎細胞激素以及抗發炎細胞激素之代表,先以此2因子對於預後準確度進行評估,再以中藥作為治療之驗證工具。研究方法為老鼠於麻醉後先由尾動脈採血測得介白素6與介白素10之基準值,之後進行盲腸結紮及穿孔手術引發老鼠敗血症,第一部分為預後評估:35隻老鼠於手術後趨於平穩之基礎心跳倍率稱為1倍,在之後逐漸增加之心跳倍率(1.15倍、1.25倍以及1.35倍)下進行抽血檢測介白素6與介白素10,並觀察其3天存活率,將造型成功者依照存活或死亡分為兩組,以此兩細胞激素為因子做預後鑑別方程式,評估其準確度;第二部分為單味中藥石膏治療:72隻老鼠隨機分為3組,在手術後6小時分別給予高劑量石膏(2.7 cc/kg)、低劑量石膏(1.35 cc/kg)以及生理食鹽水(2.7 cc/kg),評估其3天存活率,確定石膏較佳作用劑量之後,再取21隻老鼠隨機分為治療組(n=8)、控制組(n=8)與空白對照組(n=5),在手術後6小時分別給予石膏以及生理食鹽水,並且每4小時量測血壓與心跳以及抽血檢測介白素6與介白素10之數值;第三部分為中藥複方白虎湯治療:12隻老鼠隨機分為治療組(n=6)與控制組(n=6),手術後之實驗流程與石膏相同。第一部分結果顯示:介白素6的數值在死亡組隨著心跳倍率升高,數值持續上升,而存活組在1.15倍時數值雖然升高,但到了1.25倍時,即開始下降,到了1.35倍時繼續下降;介白素10亦有類似的趨勢,所不同的是到了1.25倍以及1.35倍時數值出現持平之現象。在心跳倍率1.15倍時其預後線性鑑別方程式鑑別率為68.33%;1.25倍以及1.35倍時,鑑別率分別可高至93.33%以及95.45%;第二部分結果顯示:石膏確實能有效降低敗血證老鼠之死亡率,而且對於介白素6的濃度有明顯降低之作用,至於介白素10雖無明顯降低效果但可看出其趨勢,至於血壓與心跳則無明顯作用;第三部分結果顯示:白虎湯與石膏之不同處在於對介白素6以及介白素10的濃度均有明顯降低作用。由以上結果可知:介白素6以及介白素10此兩因子對於敗血症的死亡有著重要的影響,是敗血症導致死亡的關鍵。因此,在未來的研究上,若能同時注重發炎細胞激素與抗發炎細胞激素的動態變化趨勢,進而給予及時並且正確的治療,應是一個值得深入的方向。
AbstractThe high mortality rate of sepsis is always a difficult problem to overcome in clinic. Because the course of sepsis is developed fast and the pathophisiology turns complicated, it is difficult to control in treating. After sepsis happens, the uncontrollable cytokine storm is the important reason of patient''s death, so the change of proinflammatory and anti-inflammatory cytokines is the important key whether the condition worsens. To understand the importance of proinflammatory and anti-inflammatory cytokines in mortality, the study chooses IL-6 and IL-10 to represent proinflammatory and anti-inflammatory cytokines respectively. We use these two factors to evaluate prognosis accuracy first, and then regard traditional Chinese medicine as verification tool in treatment. The baseline values of IL-6 and IL-10 were measured by collecting blood with mouse tail artery after anesthetizing. The first part is the evaluation of prognosis: 35 Sprague-Dawley rats, weighing in between 250 to 300g, underwent cecal ligation and puncture (CLP) procedures and femoral arterial catheterization. When the heart rates reached 1.15, 1.25 and 1.35 times that of the baseline heart rate, blood sampling was done for IL-6 and IL-10. The prognosis was followed up for 3 days after CLP, and all rats were then divided into survival or non-survival groups. Finally, the relationships among survival condition, heart rate, IL-6, and IL-10 were analyzed by two-way ANOVA and multiple comparison. A linear discriminant function for the prognosis of sepsis was developed by IL-6 and IL-10 at different heart rates. Second part: the treatment of single flavor traditional Chinese medicine gypsum: Seventy-two rats were randomly assigned to three groups: control group, low dose treatment group and high dose treatment group. Sepsis was induced by cecal ligation and puncture. After 6 h, control group received normal saline 2.7cc/kg, low dose and high dose treatment groups received Gypsum 1.35cc/kg and 2.7cc/kg respectively. We observed three days for survival rate analysis. To realize the mechanism of gypsum further, we developed three groups randomly: control group (n=8), the most effective group (n=8) and sham group (n=5). Normal saline or gypsum was given subcutaneously 6h later. Inflammatory cytokines IL-6 and IL-10 were measured before CLP and every 4 h after CLP for 24 h. Blood pressure and heart rate were also measured every 4 h after CLP for 24 h. The third part: to realize the mechanism of Pai-hu-tang further, we developed two groups randomly: control group (n=6) and treatment group (n=6). Normal saline or Pai-hu-tang was given subcutaneously 6h later. Inflammatory cytokines IL-6 and IL-10 were measured before CLP and every 4 h after CLP for 24 h. Blood pressure and heart rate were also measured every 4 h after CLP for 24 h. The result shows: First part: IL-6 elevated continuously with the increment of the heart rate in the non-survival group, but in the survival group, the IL-6 elevation started to fall when the heart rate reached 1.25 times; a similar trend was found in the change of IL-10. The prediction rate was 68.33% when the heart rate reached 1.15 times, and increased to 93.33% and 95.45% when heart rates reached 1.25 and 1.35 times respectively. Second part: 3-day survival rate in both treatment groups were higher than the control group. IL-6 levels were significantly higher in the control than in the treatment group at 12 h,16 h,20 h and 24 h. Though there was no significant difference between control and treatment groups, the IL-10 levels seemed to have a decreasing trend in the treatment group. There was no obvious increase in blood pressure among these three groups. Heart rate increased by time in control and treatment groups and had significant difference with sham group at 16 h, 20 h and 24 h. Third part: IL-6 levels were significantly higher in the control than in the treatment group at 12 h, 16 h,20 h and 24 h. IL-10 levels were significantly higher in the control than in the treatment group at 16 h, 20 h and 24 h. Conclusion: IL6 and IL10 are very effective to the mortality of sepsis. They are strongly correlated to the pathogenesis and treatment of sepsis. In the future, if we can pay attention to the dynamic change of these two cytokines, it may be valuable in the treatment of sepsis.
目  錄
第一章 前言 1
第二章 文獻探討 3
壹、現代醫學有關敗血症之論述 3
一、定義 3
二、病因 4
三、病理機轉 6
四、治療 9
五、預後評估 11
貳、傳統中醫有關敗血症之論述 18
一、敗血症 18
二、嚴重敗血症 19
三、敗血性休克 22
參、溫病氣分證用藥文獻探討 24
一、氣分證各類型分析 24
二、白虎湯組成 25
三、白虎湯個別藥物探討 26
肆、敗血症動物模型討論 28
一、內毒素模型 28
二、敗血症模型 29
三、盲腸結紮以及穿孔簡介 30
第三章 材料與方法 32
壹、實驗設計 32
貳、實驗動物 32
參、實驗儀器設備與試劑 32
一、手術器具 32
二、實驗試劑 33
三、實驗儀器 34
四、中藥基源鑑定及製備 34
肆、敗血症造型成功評估 35
一、血液培養 35
二、行為觀察 35
三、心跳數值 35
四、解剖 36
伍、實驗流程 36
一、實驗前置作業 36
二、預後評估實驗 37
三、石膏實驗 38
四、白虎湯實驗 38
陸、參數之測量與分析 38
一、血壓與心跳之測量 38
二、血液樣本處理 39
三、細胞激素分析 39
柒、統計方法 40
一、預後評估 40
二、石膏治療 40
三、白虎湯治療 40
第四章 結果 41
壹、預後評估實驗 41
一、基本資料 41
二、不同心跳倍率下死亡組與存活組之介白素6濃度曲線 42
三、不同心跳倍率下死亡組與存活組之介白素10濃度曲線 43
四、不同心跳倍率下死亡組與存活組之介白素6以及介白素
10濃度散佈曲線 44
五、不同心跳倍率下之預後鑑別方程式 47
貳、石膏治療實驗 49
一、3天存活曲線 49
二、各組之血壓與心跳速率比較 49
三、各組之介白素6以及介白素10濃度比較 51
參、白虎湯治療實驗 52
一、各組之血壓與心跳速率比較 52
二、各組之介白素6以及介白素10濃度比較 54
第五章 討論 56
第六章 結論 62
參考文獻 63
附錄 76
英文摘要 80
謝辭 83
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