臺灣博碩士論文加值系統

在本實驗室過去研究中發現溫灸內關穴(PC 6)會在相對應內臟(心臟)誘導熱休克蛋白七十(Hsp70)表現量增加，並進而保護大鼠心臟避免缺血再灌流傷害。然而在這當中，溫灸作用的機轉卻仍尚未瞭解。因此，本研究的目的是為了測試我們的假設：溫灸刺激內關穴位時，是否在穴位部分產生自由基而引致其相對應內臟器官(心臟)Hsp70的表現及其功能的影響。
研究方法中，在適當麻醉的雄性Sprague-Dawley大鼠上，分別在腹結穴(SP 14)和內關穴(PC 6)施予溫灸一次，平均溫度維持在41~42℃，並各分為五個組別：正常組(0 min)、溫灸後5、15、30和60分鐘，作時間效應探討，並評估穴位下肌肉和相對應內臟(心臟)組織中，自由基產生和其調控Hsp70表現機轉之參數，包括DCFH-DA螢光分析測量以H2O2為主的活性氧分子(ROS)、各種化學分析法測量包括一氧化氮( NO)終產物(NO2─和NO3─)、脂質過氧化物(MDA)、超氧化物歧化酶(SOD)、過氧化氫酶(CAT)和麩胺基硫(GSH)、酵素免疫分析法測量環磷酸鳥苷(cGMP)、西方點墨法測量Hsp70和Nrf2的蛋白表現量；最後則以血清之生化檢測項目CK-MB與Troponin I當作評估心肌損傷的參數。
結果顯示在溫灸腹結穴(SP 14)穴位下肌肉會在溫灸後5分鐘產生ROS、NO終產物(NO2─和NO3─)和MDA，並且會依序在溫灸後5分鐘消耗SOD、溫灸後15分鐘消耗CAT和GSH。此外，在溫灸前給予L-NAME會部分阻斷溫灸誘導Hsp70在穴位下肌肉的表現。而溫灸內關穴(PC 6)在相對應內臟(心臟)可觀察到ROS、cGMP、MDA和Nrf2在溫灸後5分鐘產生，而SOD在溫灸後30分鐘增加。在血清方面，CK-MB和cTnI在溫灸後15分鐘產生。此外，在溫灸內關穴(PC 6)穴位下肌肉也可觀察到ROS、NO和Nrf2在溫灸後5分鐘產生，這也再次證實了溫灸會在穴位下肌肉刺激自由基產生的結果。
本研究的結論是溫灸內關穴(PC 6) 會分別在穴位下肌肉和相對應內臟(心臟)刺激自由基產生，，而且在溫灸前給予L-NAME抑制NO產生會部分阻斷溫灸誘導Hsp70在穴位下肌肉的表現。因此可以間接解釋溫灸調控心臟Hsp70表現達到保護心臟功能的機轉。

Local somatothermal stimulation (LSTS) was demonstrated to increase the expression of myocardial heat shock protein 70 (Hsp70) and protected rat hearts against ischemia-reperfusion injury. However, the exact mechanisms of how LSTS works remains unknown. The aim of this study was to test the hypothesis that LSTS at PC 6 induced free radicals formation in the muscle beneath acupoint and subsequently induced Hsp70 expression in the heart.
LSTS was applied 0.5 cm above and onto acupoint Fujie (SP 14) and Neiguan (PC 6), respectively. Under adequate anesthesia, male Sprague–Dawley rat, were treated with LSTS for 1 dose. Following time intervals of 0 min (no treatment),5-, 15-, 30- and 60- min, respectively, reactive oxygen species (ROS) were measured by 2′,7′-dichlorofluorescin diacetate (DCFH-DA) fluorometric assay. Nitrate and nitrite (nitric oxide (NO) end products), malondialdehyde (MDA) , cyclic guanosine monophosphate (cGMP) and free radical scavengers were measured by commercial available kits. Western blot analysis for Hsp70 and Nuclear factor erythroid 2–related factor 2 (Nrf2) in the muscle and the heart were also analyzed; the parameters such as serum creatine kinase-MB isoenzyme (CK-MB) and cardiac tropnin I (cTnI) were measured to evaluate the myocardial injury.
The results showed that the generation of ROS, NO end products (nitrate and nitrite) and MDA in 5 min-, the consumption of superoxide dismutase (SOD) in 5 min-, and the consumption of catalase (CAT) and glutathione (GSH) in 15 min- after the application of LSTS suggested that there were free radicals formation in the muscle beneath the acupoint area. The LSTS-induced Hsp70 expression in the muscle were partially blocked by L-NAME treatment. Besides, the formation of ROS, MDA, cGMP and Nrf2 in 5 min- , the increase of serum CK-MB and cTnI in 15 min- and subsequently the generation of SOD in 30 min- suggested that there were free radicals formation in the heart after LSTS treatment on left PC 6. Finally, There were also ROS, NO end products (nitrate and nitrite) and Nrf2 expression in 5 min- after LSTS were noticed in the muscle beneath acupoint PC 6.
We concluded that LSTS induced free radicals formation in the muscle beneath the acupoint PC 6 and, the corresponding organ–heart, which might explain the possible mechanisms of LSTS-induced myocardial protection in rats.

目次 Ⅰ
表次及圖次 Ⅸ
中文摘要 ⅩⅢ
英文摘要 ⅩⅤ
第一章 緒論 1
第一節 自由基之概念 2
一、 自由基的產生 3
1. 活性氧分子 (Reactive oxygen species) 3
2. 活性氮分子(Reactive nitrogen species) 7
二、 自由基的氧化傷害 10
三、 自由基與缺血再灌流傷害的關係 12
四、 對抗自由基的防衛機制 13
五、 自由基調控生理反應 15
第二節 熱休克蛋白之介紹及誘發因素 16
一、 熱休克蛋白的功能與調控機制 18
二、 熱休克蛋白家族 20
三、 熱休克蛋白與自由基之關係 23
第三節 中醫觀點 26
一、 灸療的起源與發展 26
二、 針灸穴位相對的特異性 28
1. 中醫古典文獻之描述 28
2. 針灸近代相關之研究 30
三、 溫灸與局部熱效應之關係 31
第四節 本實驗室之溫灸研究成果 32
一、 溫灸誘導一氧化氮釋放對生理功能影響 32
二、 溫灸誘導熱休克蛋白產生對內臟的保護作用 33
第五節 研究動機與目的 35
第二章 實驗材料及方法 36
第一節 實驗動物 37
第二節 動物麻醉 37
第三節 實驗儀器 、藥品試劑與手術器械 37
一、 實驗儀器 37
二、 化學藥品及試劑 39
三、 手術器械 44
第四節 針灸參數的選擇 44
一、 穴位的選擇及解剖定位 44
二、 溫灸的方法與模型之建立 45
1. 溫灸參數科學化 45
2. 溫灸參數 46
3. 劑量效應與時間效應的選擇 47
第五節 實驗設計 47
一、 實驗設計及分組 47
1. 先驅實驗 47
2. 溫灸腹結穴（SP 14）在局部肌肉和心臟產生
自由基之時間效應實驗 48
3. 溫灸腹結穴（SP 14）在局部肌肉一氧化氮影響
熱休克蛋白七十表現量實驗 48
4. 溫灸內關穴（PC 6）在局部肌肉和相對應內臟
(心臟)產生自由基之時間效應實驗 49
第六節 評估方法 50
一、 2′,7′-dichlorodihydrofluorescein diacetate
(DCFH-DA)螢光分析法 50
二、 Nitrate and Nitrite (NO end products) 分析法 52
三、 Malondialdehyde (MDA) 分析法 54
四、 Superoxide Dismutase (SOD) 分析法 55
五、 Catalase (CAT) 分析法 56
六、 Glutathione (GSH) 分析法 57
七、 Cyclic guanosine monophosphate (cGMP) 分析法 59
八、 血清生化檢驗Creatine kinase-MB isoenzyme
(CK-MB)和cardiac Tropnin I (cTnI) 60
九、 Heat shock protein 70 (Hsp70)和Nuclear factor
erythroid 2-related factor 2 (Nrf2)表現量測定 62
1. 肌肉和心臟組織的處理 62
2. 蛋白質定量 62
3. 蛋白質電泳分離 63
4. 西方點墨法 (Western blotting) 64
第七節 實驗數據處理和統計方法 65
第三章 結果 66
第一節 溫灸腹結穴（SP 14）在局部肌肉刺激自由基產生
之時間效應 67
一、 溫灸腹結穴（SP 14）在局部肌肉DCFH-DA
之螢光變化 67
二、 溫灸腹結穴（SP 14）在局部肌肉之NO終產物
(NO2─＋NO3─)濃度變化 67
三、 溫灸腹結穴（SP 14）在局部肌肉之MDA濃度變化 68
四、 溫灸腹結穴（SP 14）在局部肌肉之自由基清除者：
SOD、CAT活性與GSH濃度變化 68
第二節 溫灸腹結穴（SP 14）在局部肌肉誘發NO影響Hsp70
表現量變化 70
第三節 溫灸內關穴（PC 6）在相對應內臟(心臟)刺激自由基
產生之時間效應 70
一、 溫灸內關穴（PC 6）在相對應內臟(心臟)DCFH-DA
之螢光變化 70
二、 溫灸內關穴（PC 6）在相對應內臟(心臟)之NO
終產物(NO2─＋NO3─)濃度變化 71
三、 溫灸內關穴（PC 6）在相對應內臟(心臟)之cGMP
濃度變化 71
四、 溫灸內關穴（PC 6）在相對應內臟(心臟)之MDA
濃度變化 72
五、 溫灸內關穴（PC 6）在相對應內臟(心臟)之自由基
清除者：SOD、CAT活性與GSH濃度變化 72
六、 溫灸內關穴（PC 6）對心肌的損傷指數之影響：
CK-MB、cTnI之濃度變化 74
七、 溫灸內關穴（PC 6）在相對應內臟(心臟)之Nrf2
表現量變化 75
第四節 溫灸內關穴（PC 6）在局部肌肉刺激自由基產生
之時間效應 75
一、 溫灸內關穴（PC 6）在局部肌肉DCFH-DA
之螢光變化 75
二、 溫灸內關穴（PC 6）在局部肌肉之NO終產物
(NO2─＋NO3─)濃度變化 76
三、 溫灸內關穴（PC 6）在局部肌肉之Nrf2表現量變化 76
第五節 溫灸腹結穴（SP 14）在心臟刺激自由基產生之
時間效應 76
一、溫灸腹結穴（SP 14）在心臟DCFH-DA之螢光變化 77
二、溫灸腹結穴（SP 14）在心臟之cGMP濃度變化 77
第四章 討論 78
第一節 實驗方法之討論 79
一、 麻醉藥物的使用 79
二、 實驗模型的探討 79
1. 溫灸參數的採用與實施 79
2. 穴位的選擇與其相關性 80
3. 正向控制組結果的探討 81
三、活性氧分子(ROS)分析方法的探討 81
第二節 溫灸刺激自由基產生之時間效應探討 81
一、 時間點的設定 81
二、溫灸腹結穴（SP 14）在局部肌肉刺激自由基產生
之時間效應探討 82
三、溫灸內關穴（PC 6）在局部肌肉和相對應內臟
(心臟)刺激自由基產生之時間效應探討 84
四、溫灸腹結穴（SP 14）在心臟刺激自由基產生之
時間效應探討 86
第三節 與其他熱療比較的異同點 87
一、 相異點 87
1. 方法 87
2. 前置處理和觀察時間 87
3. 機轉層面 88
二、 相同點 88
1. 機轉層面 88
2. 功能層面 89
第四節 體表內臟反射(somato-visceral reflex) 90
第五節 溫灸內關穴（PC 6）調節相對應內臟(心臟)功能
機轉探討 91
第六節 臨床應用與發展 93
第七節 未來與展望 94
第五章 結論 95
第六章 參考文獻 97
圖表附錄 115

1. Abdel-Wahab MH, Arafa HM, El-Mahdy MA, Abdel-Naim AB. Potential protective effect of melatonin against dibromoacetonitrile -induced oxidative stress in mouse stomach. Pharmacol Res. 2002;46:287-93.
2. Adelman R, Saul RL, Ames BN. Oxidative damage to DNA: relation to species metabolic rate and life span. Proc Natl Acad Sci U S A. 1988;85:2706-8.
3. Aikens J, Dix TA. Perhydroxyl radical (HOO.) initiated lipid peroxidation. The role of fatty acid hydroperoxides. J Biol Chem. 1991;266:15091-8.
4. Arnaud C, Joyeux M, Garrel C, Godin-Ribuot D, Demenge P, Ribuot C. Free-radical production triggered by hyperthermia contributes to heat stress-induced cardioprotection in isolated rat hearts. Br J Pharmacol. 2002;135:1776-82.
5. Babior BM. NADPH oxidase. Curr Opin Immunol. 2004;16:42-7.
6. Banerjee Mustafi S, Chakraborty PK, Dey RS, Raha S. Heat stress upregulates chaperone heat shock protein 70 and antioxidant manganese superoxide dismutase through reactive oxygen species (ROS), p38MAPK, and Akt. Cell Stress Chaperones. 2009.
7. Basaga HS. Biochemical aspects of free radicals. Biochem Cell Biol. 1990;68:989-98.
8. Bauman JW, Liu J, Klaassen CD. Production of metallothionein and heat-shock proteins in response to metals. Fundam Appl Toxicol. 1993;21:15-22.
9. Beckman JS, Beckman TW, Chen J, Marshall PA, Freeman BA. Apparent hydroxyl radical production by peroxynitrite: implications for endothelial injury from nitric oxide and superoxide. Proc Natl Acad Sci U S A. 1990;87:1620-4.
10. Bellmann K, Jäättelä M, Wissing D, Burkart V, Kolb H. Heat shock protein hsp70 overexpression confers resistance against nitric oxide. FEBS Lett. 1996;391:185-8.
11. Bennett MR. Non-adrenergic non-cholinergic (NANC) transmission to smooth muscle: 35 years on. Prog Neurobiol. 1997;52:159-95.
12. Bergendi L, Benes L, Duracková Z, Ferencik M. Chemistry, physiology and pathology of free radicals. Life Sci. 1999;65:1865-74.
13. Bolli R. The late phase of preconditioning. Circ Res. 2000;87:972-83.
14. Brand MD. The proton leak across the mitochondrial inner membrane. Biochem Biophys Acta. 1990; 1018: 128-133.
15. Bukau B, Horwich AL. The Hsp70 and Hsp60 chaperone machines. Cell.1998;92:351-66.
16. Buttke TM, Sandstrom PA. Oxidative stress as a mediator of apoptosis. Immunol Today. 1994;15:7-10.
17. Cadenas E. Basic mechanisms of antioxidant activity. Biofactors. 1997;6:391-7.
18. Cadet J, Berger M. Radiation-induced decomposition of the purine bases within DNA and related model compounds. Int J Radiat Biol Relat Stud Phys Chem Med. 1985;47:127-43.
19. Cao Q, Liu J, Wei Y, Han Z. Studies on the interrelationships between some acupoints on pericardial channel and heart. Zhen Ci Yan Jiu. 1990; 15: 35-9.
20. Carr AC, McCall MR, Frei B. Oxidation of LDL by myeloperoxidase and reactive nitrogen species: reaction pathways and antioxidant protection. Arterioscler Thromb Vasc Biol. 2000;20:1716-23.
21. Chirico WJ, Waters MG, Blobel G. 70K heat shock related proteins stimulate protein translocation into microsomes. Nature. 1988;332:805-10.
22. Chiueh CC. Neuroprotective properties of nitric oxide. Ann N Y Acad Sci. 1999;890:301-11.
23. Chiu JH, Lui WY, Chen YL, Hong CY. Local somatothermal stimulation inhibits the motility of sphincter of Oddi in cats, rabbits and humans through nitrergic neural release of nitric oxide. Life Sci. 1998;63:413-28.
24. Chiu JH, Tsou MT, Tung HH, Tai CH, Tsai SK, Chih CL, Lin JG, Wu CW. Preconditioned somatothermal stimulation on median nerve territory increases myocardial heat shock protein 70 and protects rat hearts against ischemia-reperfusion injury. J Thorac Cardiovasc Surg. 2003;125:678-85.
25. Cho ZH, Chung SC, Jones JP, Park JB, Park HJ, Lee HJ, Wong EK, Min BI. New findings of the correlation between acupoints and corresponding brain cortices using functional MRI. Proc Natl Acad Sci USA. 1998; 95: 2670-3.
26. Commoner B, Townsend J, Pake GE. Free radicals in biological materials. Nature. 1954;174:689-91.
27. Craig EA, Gambill BD, Nelson RJ. Heat shock proteins: molecular chaperones of protein biogenesis. Microbiol Rev. 1993;57:402-14.
28. Craig EA, Kramer J, Kosic-Smithers J. SSC1, a member of the 70-kDa heat shock protein multigene family of Saccharomyces cerevisiae, is essential for growth. Proc Natl Acad Sci U S A. 1987;84:4156-60.
29. Darley-Usmar V, Wiseman H, Halliwell B. Nitric oxide and oxygen radicals: a question of balance. FEBS Lett. 1995;369:131-5.
30. Deisseroth A, Dounce AL. Catalase: Physical and chemical properties, mechanism of catalysis, and physiological role. Physiol Rev. 1970;50:319-75.
31. Dennery PA, Rodgers PA, Lum MA, Jennings BC, Shokoohi V. Hyperoxic regulation of lung heme oxygenase in neonatal rats. Pediatr Res. 1996;40:815-21.
32. Dröge W. Free radicals in the physiological control of cell function. Physiol Rev. 2002;82:47-95.
33. Ewing JF, Raju VS, Maines MD. Induction of heart heme oxygenase-1 (HSP32) by hyperthermia: possible role in stress-mediated elevation of cyclic 3':5'-guanosine monophosphate. J Pharmacol Exp Ther. 1994;271:408-14.
34. Ferdinandy P, Schulz R. Nitric oxide, superoxide, and peroxynitrite in myocardial ischaemia-reperfusion injury and preconditioning. Br J Pharmacol. 2003;138:532-43.
35. Fialkow L, Wang Y, Downey GP. Reactive oxygen and nitrogen species as signaling molecules regulating neutrophil function. Free Radic Biol Med. 2007;42:153-64.
36. Flanagan SW, Moseley PL, Buettner GR. Increased flux of free radicals in cells subjected to hyperthermia: detection by electron paramagnetic resonance spin trapping. FEBS Lett. 1998;431:285-6.
37. Fridovich I. Superoxide radical and superoxide dismutases. Annu Rev Biochem. 1995;64:97-112.
38. Fu JR, Liu WL, Zhou JF, Sun HY, Xu HZ, Luo L, Zhang H, Zhou YF. Sonic hedgehog protein promotes bone marrow-derived endothelial progenitor cell proliferation, migration and VEGF production via PI 3-kinase/Akt signaling pathways. Acta Pharmacol Sin. 2006;27:685-93.
39. Gamaley IA, Klyubin IV. Roles of reactive oxygen species: signaling and regulation of cellular functions. Int Rev Cytol. 1999;188:203-55.
40. Gerschman R, Gilbert D, Nye SW, Dwyer P, Fenn WO. Oxygen poisoning and x-irradiation: a mechanism in common. Science. 1954;119:623-6.
41. Ghafourifar P, Cadenas E. Mitochondrial nitric oxide synthase. Trends Pharmacol Sci. 2005;26:190-5.
42. Gomes A, Fernandes E, Lima JL. Fluorescence probes used for detection of reactive oxygen species. J Biochem Biophys Methods. 2005;65:45-80.
43. Granger DN, Rutili G, McCord JM. Superoxide radicals in feline intestinal ischemia. Gastroenterology. 1981; 81: 22-9.
44. Gualtieri M, Mantecca P, Cetta F, Camatini M. Organic compounds in tire particle induce reactive oxygen species and heat-shock proteins in the human alveolar cell line A549. Environ Int. 2008;34:437-42.
45. Gupta SC, Siddique HR, Mathur N, Vishwakarma AL, Mishra RK, Saxena DK, Chowdhuri DK. Induction of hsp70, alterations in oxidative stress markers and apoptosis against dichlorvos exposure in transgenic Drosophila melanogaster: modulation by reactive oxygen species. Biochim Biophys Acta. 2007;1770:1382-94.
46. Gutteridge JM, Halliwell B. Reoxygenation injury and antioxidant protection: a tale of two paradoxes. Arch Biochem Biophys. 1990;283:223-6.
47. Hall DM, Buettner GR, Oberley LW, Xu L, Matthes RD, Gisolfi CV. Mechanisms of circulatory and intestinal barrier dysfunction during whole body hyperthermia. Am J Physiol Heart Circ Physiol. 2001;280:H509-21.
48. Halliwell B. Free radicals, antioxidants, and human disease: curiosity, cause, or consequence? Lancet. 1994;344:721-4.
49. Halliwell B, Gutteridge JM. Free radicals in biology and medicine (3rd ed.). Oxford University Press. 1999.
50. Han JB, Oh SD, Lee KS, Choi KS, Cho YW, Ahn HO, Bae H, Min BI. The role of the sympathetic nervous system in moxibustion -induced immunomodulation in rats. J Neuroimmunol. 2003;140:159-62.
51. Harman D. Aging: a theory based on free radical and radiation chemistry. J Gerontol. 1956;11:298-300.
52. Henle KJ, Leeper DB. Interaction of hyperthermia and radiation in CHO cells: recovery kinetics. Radiat Res. 1976;66:505-18.
53. Hua JS, Li LP, Zhu XM. Effects of moxibustion preconditioning on SOD and MDA in rats with global brain ischemia. Zhongguo Zhen Jiu. 2006;26:595-7.
54. Ishikawa T, Nakada-Moriya Y, Ando C, Tanda N, Nishida S, Minatogawa Y, Nohno T. Expression of the JNK2-alpha1 gene in the developing chick brain. Biochem Biophys Res Commun. 1997;234:489-92.
55. Ishiyama S, Hiroe M, Nishikawa T, Abe S, Shimojo T, Ito H, Ozasa S, Yamakawa K, Matsuzaki M, Mohammed MU, Nakazawa H, Kasajima T, Marumo F. Nitric oxide contributes to the progression of myocardial damage in experimental autoimmune myocarditis in rats. Circulation. 1997;95:489-96.
56. Jäättelä M. Heat shock proteins as cellular lifeguards. Ann Med. 1999;31:261-71.
57. Jäättelä M, Wissing D, Kokholm K, Kallunki T, Egeblad M. Hsp70 exerts its anti-apoptotic function downstream of caspase-3-like proteases. EMBO J. 1998;17:6124-34.
58. Janero DR. Malondialdehyde and thiobarbituric acid-reactivity as diagnostic indices of lipid peroxidation and peroxidative tissue injury. Free Radic Biol Med. 1990;9:515-40.
59. Javadpour M, Kelly CJ, Chen G, Stokes K, Leahy A, Bouchier-Hayes DJ. Thermotolerance induces heat shock protein 72 expression and protects against ischaemia-reperfusion-induced lung injury. Br J Surg. 1998;85:943-6.
60. Jiang JK, Chiu JH, Lin JK. Local somatothermal stimulation inhibits motility of the internal anal sphincter through nitrergic neural release of nitric oxide. Dis Colon Rectum. 2000;43:381-8.
61. Jiang JK, Chiu JH, Lin JK. Local thermal stimulation relaxes hypertonic anal sphincter: evidence of somatoanal reflex. Dis Colon Rectum. 1999;42:1152-9.
62. Jobgen WS, Fried SK, Fu WJ, Meininger CJ, Wu G. Regulatory role for the arginine-nitric oxide pathway in metabolism of energy substrates. J Nutr Biochem. 2006;17:571-88.
63. Joyeux-Faure M, Arnaud C, Godin-Ribuot D, Ribuot C. Heat stress preconditioning and delayed myocardial protection: what is new? Cardiovasc Res. 2003;60:469-77.
64. Joyeux M, Ribuot C, Bourlier V, Verdetti J, Durand A, Richard MJ, Godin-Ribuot D, Demenge P. In vitro antiarrhythmic effect of prior whole body hyperthermia: implication of catalase. J Mol Cell Cardiol. 1997;29:3285-92.
65. Kamata H, Hirata H. Redox regulation of cellular signalling. Cell Signal. 1999;11:1-14.
66. Kawakita K, Shinbara H, Imai K, Fukuda F, Yano T, Kuriyama K. How do acupuncture and moxibustion act? - Focusing on the progress in Japanese acupuncture research -. J Pharmacol Sci. 2006;100:443-59.
67. Keyse SM, Tyrrell RM. Heme oxygenase is the major 32-kDa stress protein induced in human skin fibroblasts by UVA radiation, hydrogen peroxide, and sodium arsenite. Proc Natl Acad Sci U S A. 1989;86:99-103.
68. Kiang JG, Tsokos GC. Heat shock protein 70 kDa: molecular biology, biochemistry, and physiology. Pharmacol Ther 1998;80:183-201.
69. Klatt P, Lamas S. Regulation of protein function by S-glutathiolation in response to oxidative and nitrosative stress. Eur J Biochem. 2000;267:4928-44.
70. Koshland DE Jr. The molecule of the year. Science. 1992;258:1861.
71. Kovacic P, Jacintho JD. Reproductive toxins: pervasive theme of oxidative stress and electron transfer. Curr Med Chem. 2001;8:863-92.
72. Kung YY, Chen FP, Hwang SJ. The different immunomodulation of indirect moxibustion on normal subjects and patients with systemic lupus erythematosus. Am J Chin Med. 2006;34:47-56.
73. Lawler JM, Song W, Demaree SR. Hindlimb unloading increases oxidative stress and disrupts antioxidant capacity in skeletal muscle. Free Radic Biol Med. 2003;35:9-16.
74. Lindquist S. The heat-shock response. Annu Rev Biochem. 1986;55:1151-91.
75. Lin KM, Lin B, Lian IY, Mestril R, Scheffler IE, Dillmann WH. Combined and individual mitochondrial HSP60 and HSP10 expression in cardiac myocytes protects mitochondrial function and prevents apoptotic cell deaths induced by simulated ischemia-reoxygenation. Circulation. 2001;103:1787-92.
76. Lin YH, Chiu JH, Tung HH, Tsou MT, Lui WY, Wu CW. Preconditioning somatothermal stimulation on right seventh intercostal nerve territory increases hepatic heat shock protein 70 and protects the liver from ischemia-reperfusion injury in rats. J Surg Res. 2001;99:328-34.
77. Liochev SI, Fridovich I. The Haber-Weiss cycle -- 70 years later: an alternative view. Redox Rep. 2002;7:55-7.
78. Liu P, Hock CE, Nagele R, Wong PY. Formation of nitric oxide, superoxide, and peroxynitrite in myocardial ischemia-reperfusion injury in rats. Am J Physiol. 1997;272:H2327-36.
79. Li W, Kong AN. Molecular mechanisms of Nrf2-mediated antioxidant response. Mol Carcinog. 2009;48:91-104.
80. Lord-Fontaine S, Averill DA. Enhancement of cytotoxicity of hydrogen peroxide by hyperthermia in chinese hamster ovary cells: role of antioxidant defenses. Arch Biochem Biophys. 1999;363:283-95.
81. Malyshev IYu, Bayda LA, Trifonov AI, Larionov NP, Kubrina LD, Mikoyan VD, Vanin AF, Manukhina EB. Cross-talk between nitric oxide and HSP70 in the antihypotensive effect of adaptation to heat. Physiol Res. 2000;49:99-105.
82. Malyshev IYu, Manukhina EB, Mikoyan VD, Kubrina LN, Vanin AF. Nitric oxide is involved in heat-induced HSP70 accumulation. FEBS Lett. 1995;370:159-62.
83. Manucha W, Vallés PG. Cytoprotective role of nitric oxide associated with Hsp70 expression in neonatal obstructive nephropathy. Nitric Oxide. 2008;18:204-15.
84. Marnett LJ. Chemistry and biology of DNA damage by malondialdehyde. IARC Sci Publ. 1999;150:17-27.
85. Marnett LJ. Peroxyl free radicals: potential mediators of tumor initiation and promotion. Carcinogenesis. 1987;8:1365-73.
86. Massey V. Activation of molecular oxygen by flavins and flavoproteins. J Biol Chem. 1994;269:22459-62.
87. McCord JM, Fridovich I. Superoxide dismutase. An enzymic function for erythrocuprein (hemocuprein). J Biol Chem. 1969;244:6049-55.
88. McCord JM. Oxygen-derived free radicals in postischemic tissue injury. New Engl J Med. 1985; 312: 159-63.
89. Meldrum KK, Burnett AL, Meng X, Misseri R, Shaw MB, Gearhart JP, Meldrum DR. Liposomal delivery of heat shock protein 72 into renal tubular cells blocks nuclear factor-kappaB activation, tumor necrosis factor-alpha production, and subsequent ischemia-induced apoptosis. Circ Res. 2003;92:293-9.
90. Mestril R, Dillmann WH. Heat shock proteins and protection against myocardial ischemia. J Mol Cell Cardiol. 1995;27:45-52.
91. Michels AA, Kanon B, Konings AW, Ohtsuka K, Bensaude O, Kampinga HH. Hsp70 and Hsp40 chaperone activities in the cytoplasm and the nucleus of mammalian cells. J Biol Chem. 1997;272:33283-9.
92. Mittal CK, Murad F. Activation of guanylate cyclase by superoxide dismutase and hydroxyl radical: a physiological regulator of guanosine 3',5'-monophosphate formation. Proc Natl Acad Sci U S A. 1977;74:4360-4.
93. Mi YQ, Wu YC, Chen Y. Effects of warm needle moxibustion on nerve root local inflammatory factors (NOS and CGRP) in the lumbar nerve root compress model rats. Zhongguo Zhen Jiu. 2009;29:48-52.
94. Morano KA, Thiele DJ. Heat shock factor function and regulation in response to cellular stress, growth, and differentiation signals. Gene Expr. 1999;7:271-82.
95. Morimoto RI. Regulation of the heat shock transcriptional response: cross talk between a family of heat shock factors, molecular chaperones, and negative regulators. Genes Dev. 1998;12:3788-96.
96. Morimoto RI, Sarge KD, Abravaya K. Transcriptional regulation of heat shock genes. A paradigm for inducible genomic responses. J Biol Chem. 1992;267:21987-90.
97. Muller FL, Liu Y, Van Remmen H. Complex III releases superoxide to both sides of the inner mitochondrial membrane. J Biol Chem. 2004;279:49064-73.
98. Munro JM, Cotran RS. The pathogenesis of atherosclerosis: atherogenesis and inflammation. Lab Invest. 1988;58:249-61.
99. Murrant CL, Reid MB. Detection of reactive oxygen and reactive nitrogen species in skeletal muscle. Microsc Res Tech. 2001;55:236-48.
100. Nathan DF, Lindquist S. Mutational analysis of Hsp90 function: interactions with a steroid receptor and a protein kinase. Mol Cell Biol. 1995;15:3917-25.
101. Ni D, Gu Q, Hu HZ, Gao N, Zhu MX, Lee LY. Thermal sensitivity of isolated vagal pulmonary sensory neurons: role of transient receptor potential vanilloid receptors. Am J Physiol Regul Integr Comp Physiol. 2006;291:R541-50.
102. Nishimura H, Emoto M, Kimura K, Yoshikai Y. Hsp70 protects macrophages infected with Salmonella choleraesuis against TNF-alpha-induced cell death. Cell Stress Chaperones. 1997;2:50-9.
103. Nollen EA, Brunsting JF, Roelofsen H, Weber LA, Kampinga HH. In vivo chaperone activity of heat shock protein 70 and thermotolerance. Mol Cell Biol. 1999;19:2069-79.
104. Nowak TS Jr, Ikeda J, Nakajima T. 70-kDa heat shock protein and c-fos gene expression after transient ischemia. Stroke. 1990;21:107-11.
105. Ohtsuka K, Masuda A, Nakai A, Nagata K. A novel 40-kDa protein induced by heat shock and other stresses in mammalian and avian cells. Biochem Biophys Res Commun. 1990;166:642-7.
106. Otterbein LE, Choi AM. Heme oxygenase: colors of defense against cellular stress. Am J Physiol Lung Cell Mol Physiol. 2000;279:L1029-37.
107. Pan PJ, Chan RC, Yang AH, Chou CL, Cheng YF, Chiu JH. Protective effects of preconditioned local somatothermal stimulation on neuromuscular plasticity against ischemia--reperfusion injury in rats. J Orthop Res. 2008;26:1670-4.
108. Pastor N, Weinstein H, Jamison E, Brenowitz M. A detailed interpretation of OH radical footprints in a TBP-DNA complex reveals the role of dynamics in the mechanism of sequence-specific binding. J Mol Biol. 2000;304:55-68.
109. Paulus WJ, Bronzwaer JG. Nitric oxide's role in the heart: control of beating or breathing? Am J Physiol Heart Circ Physiol. 2004;287:H8-13.
110. Piantadosi CA, Carraway MS, Babiker A, Suliman HB. Heme oxygenase-1 regulates cardiac mitochondrial biogenesis via Nrf2-mediated transcriptional control of nuclear respiratory factor-1. Circ Res. 2008;103:1232-40.
111. Pirkkala L, Nykänen P, Sistonen L. Roles of the heat shock transcription factors in regulation of the heat shock response and beyond. FASEB J. 2001;15:1118-31.
112. Plumier JC, Ross BM, Currie RW, Angelidis CE, Kazlaris H, Kollias G, Pagoulatos GN. Transgenic mice expressing the human heat shock protein 70 have improved post-ischemic myocardial recovery. J Clin Invest. 1995;95:1854-60.
113. Poss KD, Tonegawa S. Reduced stress defense in heme oxygenase 1-deficient cells. Proc Natl Acad Sci U S A. 1997;94:10925-30.
114. Puglia CD, Powell SR. Inhibition of cellular antioxidants: a possible mechanism of toxic cell injury. Environ Health Perspect. 1984;57:307-11.
115. Qi H, Menzel R, Tse-Dinh YC. Effect of the deletion of the sigma 32-dependent promoter (P1) of the Escherichia coli topoisomerase I gene on thermotolerance. Mol Microbiol. 1996;21:703-11.
116. Rabindran SK, Giorgi G, Clos J, Wu C. Molecular cloning and expression of a human heat shock factor, HSF1. Proc Natl Acad Sci U S A. 1991;88:6906-10.
117. Rastaldo R, Pagliaro P, Cappello S, Penna C, Mancardi D, Westerhof N, Losano G. Nitric oxide and cardiac function. Life Sci. 2007;81:779-93.
118. Rawls SM, Tallarida RJ, Gray AM, Geller EB, Adler MW. L-NAME (N omega-nitro-L-arginine methyl ester), a nitric-oxide synthase inhibitor, and WIN 55212-2 [4,5-dihydro-2-methyl-4 (4-morpholinylmethyl)-1-(1-naphthalenyl-carbonyl)-6H-pyrrolo[3,2,1ij]quinolin-6-one], a cannabinoid agonist, interact to evoke synergistic hypothermia. J Pharmacol Exp Ther. 2004;308:780-6.
119. Riabowol KT, Mizzen LA, Welch WJ. Heat shock is lethal to fibroblasts microinjected with antibodies against hsp70. Science. 1988;242:433-6.
120. Ritossa FM. A new puffing pattern induced by a temperature shock and DNP in Drosophila. Exp Biol. 1962;18:571-3.
121. Roshchupkin DI, Murina MA. Free-radical and cyclooxygenase -catalyzed lipid peroxidation in membranes of blood cells under UV irradiation. Membr Cell Biol. 1998;12:279-86.
122. Rotrosen D, Gallin JI. Disorders of phagocyte function. Annu Rev Immunol. 1987;5:127-50.
123. Samali A, Holmberg CI, Sistonen L, Orrenius S. Thermotolerance and cell death are distinct cellular responses to stress: dependence on heat shock proteins. FEBS Lett. 1999;461:306-10.
124. Sen CK, Packer L. Antioxidant and redox regulation of gene transcription. FASEB J. 1996;10:709-20.
125. Siddique HR, Gupta SC, Mitra K, Bajpai VK, Mathur N, Murthy RC, Saxena DK, Chowdhuri DK. Adverse effect of tannery waste leachates in transgenic Drosophila melanogaster: role of ROS in modulation of Hsp70, oxidative stress and apoptosis. J Appl Toxicol. 2008;28:734-48.
126. Singh I, Kremser K, Ghosh B, Singh AK, Pai S. Abnormality in translational regulation of catalase expression in disorders of peroxisomal biogenesis. J Neurochem. 1996;67:2373-8.
127. Thomas CE, Morehouse LA, Aust SD. Ferritin and superoxide dependent lipid peroxidation. J Biol Chem. 1985; 260: 3275-80.
128. Turrens JF, Freeman BA, Levitt JG, Crapo JD. The effect of hyperoxidia on superoxide production by lung submitochondrial particles. Arch Biochem Biophys. 1982; 217: 401-10.
129. Valko M, Leibfritz D, Moncol J, Cronin MT, Mazur M, Telser J. Free radicals and antioxidants in normal physiological functions and human disease. Int J Biochem Cell Biol. 2007;39:44-84.
130. Valko M, Morris H, Cronin MT. Metals, toxicity and oxidative stress. Curr Med Chem. 2005;12(10):1161-208.
131. Wang CC, Chen F, Kim E, Harrison LE. Thermal sensitization through ROS modulation: a strategy to improve the efficacy of hyperthermic intraperitoneal chemotherapy. Surgery. 2007;142:384-92.
132. Weisiger RA, Fridovich I. Superoxide dismutase. Organelle specificity. J Biol Chem. 1973;248:3582-92.
133. Welch WJ. Heat shock proteins functioning as molecular chaperones: their roles in normal and stressed cells. Philos Trans R Soc Lond B Biol Sci. 1993;339:327-33.
134. Westerheide SD, Morimoto RI. Heat shock response modulators as therapeutic tools for diseases of protein conformation. J Biol Chem. 2005;280:33097-100.
135. Wiegant FA, Souren JE, van Wijk R. Stimulation of survival capacity in heat shocked cells by subsequent exposure to minute amounts of chemical stressors; role of similarity in hsp-inducing effects. Hum Exp Toxicol. 1999;18:460-70.
136. Woo AY, Cheng CH, Waye MM. Baicalein protects rat cardiomyocytes from hypoxia/reoxygenation damage via a prooxidant mechanism. Cardiovasc Res. 2005;65:244-53.
137. Xu Q, Hu Y, Kleindienst R, Wick G. Nitric oxide induces heat-shock protein 70 expression in vascular smooth muscle cells via activation of heat shock factor 1. J Clin Invest. 1997;100:1089-97.
138. Yamada N, Yamaya M, Okinaga S, Lie R, Suzuki T, Nakayama K, Takeda A, Yamaguchi T, Itoyama Y, Sekizawa K, Sasaki H. Protective effects of heme oxygenase-1 against oxidant-induced injury in the cultured human tracheal epithelium. Am J Respir Cell Mol Biol. 1999;21:428-35.
139. Yamashita N, Hoshida S, Taniguchi N, Kuzuya T, Hori M. Whole-body hyperthermia provides biphasic cardioprotection against ischemia/reperfusion injury in the rat. Circulation. 1998;98:1414-21.
140. Yin Y, Terauchi Y, Solomon GG, Aizawa S, Rangarajan PN, Yazaki Y, Kadowaki T, Barrett JC. Involvement of p85 in p53-dependent apoptotic response to oxidative stress. Nature. 1998;391:707-10.
141. Zuo L, Christofi FL, Wright VP, Liu CY, Merola AJ, Berliner LJ, Clanton TL. Intra- and extracellular measurement of reactive oxygen species produced during heat stress in diaphragm muscle. Am J Physiol Cell Physiol. 2000;279:C1058-66.
142. 王金金、蔣松鶴、李莎莎、蔣培偉，十二經脈四肢遠端穴位與脊髓神經節段支配規律的關係，中華中醫藥學刊，2008；6：1272-3。
143. 張紅星、黃國付、周利、吳文莉、劉悅平，內關心臟相關性臨床與實驗研究進展，中華中醫藥學刊，2007；7：1413-6。
144. 陳勇利，局部熱弛張兔子總膽管奧迪氏括約肌之機轉探討：一氧化氮的角色，國立陽明大學傳統醫藥學研究所碩士論文，1997。
145. 施松杉，針灸對大白鼠敗血症之免疫調控與機轉之探討，國立陽明大學傳統醫學研究所碩士論文，1999。
146. 童華雄，探討溫灸大白鼠體表穴位及相對應內臟器官熱休克蛋白七十之表現，國立陽明大學傳統醫學研究所碩士論文，2000。
147. 陳應明，溫灸大白鼠體表穴穴對腎臟微循環之影響，國立陽明大學傳統醫藥學研究所碩士論文，2002。
148. 鄒孟婷，針灸大白鼠內關穴對心肌保護之機轉探討，國立陽明大學傳統醫學研究所博士論文，2004。
149. 謝政橘，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，國立陽明大學傳統醫學研究所碩士論文，2005。
150. 黃維三，針炙科學，國立編譯館出版，台北，1985。
151. 林昭庚、鄢良，針灸醫學史，中國中醫藥出版社，北京，1995。
152. 林昭庚，新針炙大成，中國中醫藥學院針灸研究中心出版，台北，1996。
153. 林昭庚，針灸學，知音出版社，台北，2009。