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研究生:謝政橘
研究生(外文):Cheng-Chu Hsieh
論文名稱:整合中西醫學對於大白鼠肝臟缺血再灌流損傷保護效果之研究
論文名稱(外文):Effects of Integrated Traditional Chinese Medicine and Western Medicine against Ischemia-Reperfusion Injury of the Liver in Rats
指導教授:邱仁輝邱仁輝引用關係
指導教授(外文):Jen-Hwey Chiu
學位類別:碩士
校院名稱:國立陽明大學
系所名稱:傳統醫藥學研究所
學門:醫藥衛生學門
學類:藥學學類
論文種類:學術論文
論文出版年:2005
畢業學年度:93
語文別:中文
論文頁數:102
中文關鍵詞:缺血再灌流損傷電針溫灸前置處理日月穴期門穴
外文關鍵詞:Ischemia-reperfusion injuryelectro-acupuncturelocal somatothermal stimulationpreconditionRiyue (GB 24)Qimen (LR 14)
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臨床上肝臟的缺血再灌流損傷可能發生在肝臟鈍傷、肝臟移植或肝臟腫瘤的切除等的情況。許多研究顯示肝臟缺血再灌流的損傷,會引起肝臟的衰竭,而這是主要手術後引起院內死亡的原因。所以,探討治療或預防策略來對抗肝臟缺血再灌流所造成的損傷是相當重要的。
在本研究中主要有兩個目的,第一是評估電針前置處理期門穴(LR 14)和日月穴(GB 24)是否可以降低大白鼠肝臟缺血再灌流的損傷。第二則是評估中西醫結合之概念,將溫灸前置處理期門穴(LR 14),併用現在西醫之用藥N-acetylcysteine (NAC)或/和Prostaglandin E1 (PGE1)是否可以降低大白鼠肝臟缺血再灌流的損傷。
研究方法中,在電針前置處理的實驗方面,共分為五組,每組均有缺血再灌流的損傷:第一組是無給予電針的前置處理組(I/R alone);第二組是手術前30分鐘電針的前置處理組(EA-0.5h+I/R);第三組是手術前2小時電針的前置處理組(EA-2h+I/R);第四組是手術前12小時電針的前置處理組(EA-12h+I/R);第五組是手術前36小時,每12小時電針1次,共3次電針的前置處理組(EA-36h+I/R)。
在溫灸前置處理併用NAC或/和PGE1的實驗方面,共分為九組,除第一組不做缺血再灌流損傷以外,其餘八組均有缺血再灌流的損傷:第一組是不給予任何處理組(Normal);第二組是單獨的缺血再灌流組(I/R alone);第三組是手術前溫灸的前置處理組(LSTS+I/R);第四組是手術時給予NAC(150 mg/kg)的治療組(NAC+I/R); 第五組是手術時給予PGE1(0.05μg/kg)的治療組(PGE1+I/R);第六組是手術時給予NAC和PGE1的治療組(NAC+PGE1+I/R);第七組是手術前溫灸的前置處理合併手術時給予NAC的治療組(LSTS+NAC+I/R);第八組是手術前溫灸的前置處理合併手術時給予PGE1的治療組(LSTS+PGE1+I/R);第九組是手術前溫灸的前置處理合併手術時給予NAC和PGE1的治療組(LSTS+NAC+PGE1+I/R)。
評估肝臟損傷和其保護機轉之參數,包括細胞型態(H&E)、血清中之ALT和AST,肝臟組織內的脂質過氧化物(Malondialdehyde; MDA)、超氧化物歧化酶(Superoxide oxidase; SOD)、過氧化氫酶(Catalase; CAT)、麩胺基硫(Glutathione; GSH)、髓過氧化酶(Myeloperoxidase; MPO)及一氧化氮(Nitric oxide; NO)之含量。
結果顯示經由電針前置處理期門穴(LR 14)和日月穴(GB 24),並不能保護肝臟缺血再灌流的損傷。但是,經由單獨溫灸前置處理期門穴(LR 14),單獨使用NAC或PGE1均可達到保護肝臟缺血再灌流的損傷。另外,可以發現溫灸可以促進SOD的活性,抑制MPO的產生而達到保護肝臟缺血再灌流的損傷。在給予單獨NAC的治療方面,可以發現增加GSH的含量,促進SOD、catalase的活性,減少MPO的產生及抑制NO的產生,達到保護肝臟缺血再灌流的損傷。在給予單獨使用PGE1的治療方面,促進SOD的活性及抑制NO的產生,達到保護肝臟缺血再灌流的損傷。而在結合溫灸及NAC或/及PGE1則無法達到保護作用。
本實驗的結論是電針前置處理期門穴(LR 14)和日月穴(GB 24)並無法降低大白鼠肝臟缺血再灌流的損傷。而在溫灸前置處理期門穴(LR 14)併用NAC或PGE1方面,可以發現單獨的給予溫灸、NAC或PGE1可以降低大白鼠肝臟缺血再灌流的損傷,但合併使用時則無法降低大白鼠肝臟缺血再灌流的損傷。這對未來臨床上中西醫如何地來預防肝臟的缺血再灌流之損傷,提供了一重要的理論基礎與驗證。
Ischemia-reperfusion (I/R) injury of the liver may occur under many clinical conditions, such as hepatic trauma, hepatic transplantation or partial hepatectomy for liver tumors. There is evidence that the sequence of hepatic I/R injury causes liver injury, followed by the development of hepatic failure, which is the main cause of death after hepatetomy. Therefore, it is important to investigate the treatment or/and preventive strategies against I/R injuty of the liver.
There were two main purposes in this study. The first one was designed to evaluate the hypothesis that electro-acupuncture (EA) precondition on Qimen (LR 14) and Riyue (GB 24) attenuated I/R injury of the liver. The second one was designed to evaluate the hypothesis that by integration of traditional Chinese medicine with western medicine, local somatothermal stimulation (LSTS) precondition on Qimen (LR 14) combined with the use of N-acetylcysteine (NAC) and/or Prostaglandin E1 (PGE1) attenuated I/R injury of the liver in rats.
For the first purpose, Sprague-Dawley rats were randomly allocated into 5 groups: group 1, I/R alone; group 2, preconditioning EA on Qimen (LR 14) and Riyue (GB 24) with an interval of 30 minutes between EA and I/R injury; group 3, preconditioning EA on Qimen (LR 14) and Riyue (GB 24) with an interval of 2 hours between EA and I/R injury; group 4, preconditioning EA on Qimen (LR 14) and Riyue (GB 24) with an interval of 12 hours between EA and I/R injury; group 5, preconditioning EA on Qimen (LR 14) and Riyue (GB 24) with an interval of 12 hours, repeatedly for 3 times, followed by I/R injury. The rats were subjected to 60 minutes of hepatic ischemia followed by 60 minutes of reperfusion period.
For the second purpose, Sprague-Dawley rats were randomly allocated into 9 groups: group 1, normal; group 2, I/R alone; group 3, LSTS+I/R; group 4, NAC+I/R; group 5, PGE1+I/R; group 6, NAC+PGE1+I/R; group 7, LSTS+NAC+I/R, group 8, LSTS+PGE1+I/R; group 9, LSTS+NAC+PGE1+I/R. Preconditioning LSTS on Qimen (LR 14) with an interval of 12 hours between LSTS and I/R injury of the liver. The rats were subjected to 60 minutes of hepatic ischemia followed by 60 minutes of reperfusion period. NAC (150 mg/kg) and/or PGE1 (0.05 μg/kg) were administered alone or in combination, continuous injection, 30 minutes prior to ischemia and alone with reperfusion period.
The parameters for evaluating I/R injury of the liver, include morphology, serum alanine aminotransferase (ALT) and aspartate aminotransferase (AST), tissues levels for malondialdehyde (MDA), superoxide dismutase (SOD), catalase (CAT), glutathione (GSH), Nitric oxide (NO) and myeloperoxidase (MPO).
The results showed that precondition of EA on Qimen (LR 14) and Riyue (GB 24) did not attenuate I/R injury of the liver in rats. However, precondition of LSTS alone on Qimen (LR 14), or plus N-acetylcysteine (NAC) alone or Prostaglandin E1 (PGE1) alone attenuated I/R injury of the liver. In addition, LSTS enhanced activity of SOD as well as inhibited production of MPO. NAC alone was found to increase GSH levels, enhance activity of SOD and catalase and inhibits production of MPO and NO, and then protected the liver against I/R injury. PGE1 alone was found to enhance activity of SOD and inhibit NO production. Precondition of LSTS on Qimen (LR 14), plus N-acetylcysteine (NAC) and/or Prostaglandin E1 (PGE1) did not attenuate I/R injury of the liver.
We conclude that preconditioning EA on Qimen (LR 14) and Riyue (GB 24) does not attenuate I/R injury of the liver. LSTS, NAC or PGE1 alone, but not in combinations, protect the liver against I/R injury. This provides important information on clinical treatment strategies for I/R injury of the liver.
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