銀翹散對急性肺部發炎老鼠模式的保護作用及免疫調節之研究;Protective and Immunomodulatory effect of Gin-Gyo-San in a murine model of acute lung inflammation

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(1)ύ୯ᙴᛰεᏢύ୯ᙴᏢࣴ‫܌ز‬റγፕЎ. ࡰ Ꮴ ௲ ௤Ǻଯۘቺ ௲௤. ፕЎᚒҞ ሌᙋණჹ࡚‫ޤ܄‬೽ว‫ݹ‬ԴႵኳԄ‫ߥޑ‬ៈբҔϷխࣝፓ࿯ϐࣴ‫ز‬ Protective and Immunomodulatory effect of Gin-Gyo-San in a murine model of acute lung inflammation. ࣴ‫ز‬ғǺယৎՃ. ύ୯ᙴᛰεᏢύ୯ᙴᏢࣴ‫܌ز‬ ύ๮҇୯ΐΜϤԃѤД. i.

(2) ii.

(3) iii.

(4) Ҟᒵ ಃ΋ക. ߻‫ق‬. 1. ಃΒക. Ў᝘௖૸. 4. Ȑ΋ȑύᙴЎ᝘ӣ៝. 4. ಃ΋࿯. ‫ݹޤ‬ύᙴੰӜ‫ޑ‬ౢғᆶ‫ݮ‬ॠ. 4. ಃΒ࿯. ‫ੰݹޤ‬Ӣੰᐒ. 4. ಃΟ࿯. ύᙴჹ‫ޑݹޤ‬ϩ᛾Ϸ‫ݯ‬ᕍཷፕ. 5. ಃѤ࿯. ሌᙋණ‫ޑ‬Ў᝘ӣ៝. 6. ȐΒȑ౜жᙴᏢЎ᝘ӣ៝. ಃΟക. 9. ಃ΋࿯. ϣࢥનϷ‫ځ‬ౢғ‫ޑ‬ว‫ݹ‬ϸᔈ. ಃΒ࿯. ‫ޤ‬೽ว‫ݹ‬ϸᔈ࣬ᜢ‫ޑ‬ಒझᐟનϟಏ. 9 11. ‫׷‬਑ᆶБ‫ݤ‬. ಃ΋࿯. ‫׷‬਑. 18. 1ǵჴᡍ୏‫ނ‬. 18. 2ǵགࢉচ. 18. 3ǵ၂Ꮚᛰࠔ. 18. 4ǵ‫ל‬ᡏ. 19. 5ǵ઻‫׷‬. 20. 6ǵሺᏔ. 20. 7ǵሌᙋණ‫ޑ‬ᇙഢ. 21. ಃΒ࿯. Б‫ݤ‬. 1ǵLPS ᇨว‫ݹޤ‬Ⴕϐ୏‫ނ‬ኳԄࡌҥ. 23. 2ǵЍ਻ᆅ‫ࢱؑݰޤ‬నϐಒझᏢᔠෳ. 24. 3ǵಔᙃࢉՅϐಒझᢀჸ. 25. 4ǵNitrite ֖ໆෳ‫ۓ‬. 25. 5ǵELISA ϩ‫݋‬. 26. 6ǵਡೈқϩ‫݋‬. 27. 7ǵRNA ϩ‫݋‬. 32. 8ǵಒझ୻ᎦϷෳໆಒझғߏϷᛰ‫܄ࢥނ‬. 34. iv.

(5) 9ǵሌᙋණ‫ܭ‬ଯਏన࣬ቫ‫݋‬ϐԋҽϩ‫݋‬. 35. 10ǵTHP-1 ൂਡಒझჹ A549 ΢Ҝಒझ‫ߕޑ‬๱Κϩ‫݋‬. 36. 11ǵࢬԄಒझሺෳ‫ߕۓ‬๱ϩη. 37. 12ǵ಍ीБ‫ݤ‬. 37. ಃѤക. ่݀. ಃ΋೽ҽ ሌᙋණჹ LPS ᇨว೷ԋ‫ޤ‬ว‫ݹ‬ԴႵኳԄߥៈբҔϷ խࣝፓ࿯ᐒ‫ڋ‬ϩ‫݋‬ ಃ΋࿯ όӕᐚࡋ LPS ೷ԋ‫ޤ‬೽ཞ໾‫ޑ‬ቹៜ. 38. ಃΒ࿯ ሌᙋණჹ LPS ೷ԋ‫ޤ‬೽ཞ໾‫ޑ‬ቹៜ. 38. ಃΟ࿯ ሌᙋණჹ LPS ೷ԋ‫ޤ‬೽ཞ໾ౢғ‫਻ޑ‬ᆅϣᅖ. 42. рనϐቹៜ ಃѤ࿯ ሌᙋණჹ LPS ೷ԋ‫ޤ‬೽ཞ໾ౢғ‫ࢱؑݰޤޑ‬. 43. నύ‫ޑ‬Ոౚໆϐቹៜ ಃϖ࿯! ሌᙋණჹ LPS ᇨว‫ޑ‬ᖿϯӢη‫ޑ‬ቹៜ. 47. ಃϤ࿯ ሌᙋණჹ LPS ᇨว‫ޑ‬ว‫ݹ‬Ϸ‫ל‬ว‫ޑݹ‬ಒझᐟન. 49. ϐቹៜ ಃΎ࿯ ሌᙋණჹ LPS ೷ԋ‫ޤ‬೽ཞ໾‫ࢱؑݰޤޑ‬నύ‫ޑ‬. 51. ΋਼ϯේϷ‫ޤ‬ಔᙃύ‫ޑ‬ಒझᐟનᇨว NO ‫ޑ‬ӝԋ ሇનϐቹៜ ಃΖ࿯ ሌᙋණჹ LPS ೷ԋ‫ޤ‬೽ཞ໾ౢғ‫ޤޑ‬ಔᙃύ‫ޑ‬. 51. NF-țB Ϸ AP-1 ᙯᒵӢηϐቹៜ ಃΒ೽ϩ ሌᙋණჹ LPS ᇨวႵѮᏘಒझಒझᐟન‫ޑ‬ቹៜ ಃ΋࿯ ሌᙋණϷ‫ځ‬ಔԋϐύᛰჹ LPS ᇨวว‫ݹ‬Ⴕ‫ޑ‬. 55. ѮᏘಒझғӸ౗ϐቹៜ ಃΒ࿯ ሌᙋණϷ‫ځ‬ಔԋϐύᛰჹ LPS ᇨวว‫ޑݹ‬ಒ. 58. झᐟન TNFĮ ϐቹៜ ಃΟ࿯ ሌᙋණϷ‫ځ‬ಔԋϐύᛰჹ LPS ᇨว‫ל‬ว‫ޑݹ‬ v. 58.

(6) ಒझᐟન IL-10 ϐቹៜ ಃѤ࿯ ሌᙋණϐଯਏన࣬ቫ‫݋‬ϩ‫݋‬. 61. ಃϖ࿯ ᆘচለჹႵѮᏘಒझ‫ ޑ‬IL-10 ϩ‫ݜ‬ϐቹៜ. 61. ಃϤ࿯ ᐈҜનჹ LPSǵTNFĮ ᆶ IL-1ȕ ᇨวΓᜪ՟. 61. ѮᏘಒझ THP-1 ϷΓᜪ‫ޤ‬೽΢Ҝಒझ A549 ϐಒझᐟનϩ‫ݜ‬ϐቹៜ ಃΎ࿯ ᐈҜનჹ߻ว‫ݹ‬ಒझᐟનᇨว‫ޑ‬ಒझߕ๱ϩ. 64. ηϐቹៜ ಃΖ࿯ ᐈҜનჹ LPS ࢲϯႵ‫ޤ‬ಔᙃϷ߻ว‫ݹ‬ಒझᐟન. 67. ࢲϯ A549 ಒझਲ਼‫ ޑ‬NF-țB Ϸ AP-1 ϐቹៜ ಃΐ࿯ ᐈҜનჹ TNFĮ ᇨว‫ ޑ‬IțB Ϸ MAPK ၡ৩ϐቹៜ 69 ಃϖക. ૸ፕ. 71. ಃϤക. ่ፕ. 82. ୖԵЎ᝘. 83. मЎᄔा. 106. բ‫ޣ‬ᙁϟ. 108. vi.

(7) კҞᒵ Figure 2.1 The cytokine network between the different cells.. 13. Figure 4.1 the morphologic damage of lungs exposed to different 39 concentration of LPS. Figure 4.2 The lung morphology changes of GGS treatment.. 40. Figure 4.3 The lung morphology changes of GGS, HBX and. 41. Hesperidin treatment. Figure 4.4 The weight of lung and concentrations of proteins in. 44. BALF(I). Figure 4.5 The weight of lung and concentrations of proteins in. 45. BALF(II). Figure 4.6 The cell counts in the BALF.. 46. Figure 4.7 The tatol cell counts and neutrophil number in the. 48. BALF. Figure 4.8 Concentrations of the chemokines KC in the. 50. bronchoalveolar lavage fluid (BALF) and MIP-1 and MIP-2 in the lung tissue. Figure 4.9 Concentrations of tumor necrosis factor-alpha (TNFĮ), 52 interleukin (IL)-1ȕ, IL-6, IL-4, IL-10, and IL-12 in the bronchoalveolar lavage fluid (BALF). Figure 4.10 Concentrations of the nitrite/nitrate and iNOS.. 54. Figure 4.11 GGS Reduces NF-țB and AP-1 activation.. 56. Figure 4.12 GGS and its components effect the cell viability of. 57. mouse macrophage RAW 264.7 cell line. Figure 4.13 Concentrations of tumor necrosis factor-alpha (TNFĮ) 59 in mouse macrophage RAW 264.7. Figure 4.14 Concentrations of IL-10 in mouse macrophage RAW 60 264.7. Figure 4.15 HPLC chromatogram of the GGS. vii. 62.

(8) Figure 4.16 Concentrations of IL-10 in mouse macrophage RAW 63 264.7. Figure 4.17 Effect of HES on the expression of cytokine in lung. 65. epithelial cells A549 and macrophage-like cells THP-1. Figure 4.18 HES reduces adhesion of THP-1 cells to A549. 66. monolayers and Selective inhibition of TNF-Į or IL-1ȕ-stimulated ICAM-1 or VCAM-1 expression. Figure 4.19 Hesperidin Reduces NF-țB and AP-1 activation in. 68. Lungs Exposed to Lipopolysaccharide and in A549 cells Exposed to proinflammatory cytokine. Figure 4.20 Effect of HES on TNFĮ-induced IțB, ERK1/2, p38, or 70 JNK activation in A549 epithelial cells.. viii.

(9) ߄ԛ Table 3.1 Oligonucleotide primer designation for RT-PCR analysis 34 Table 3.2 The time and partition of solvent. 36. Table 4.1 Effect of GGS on LPS-induced cytokines.. 57. ix.

(10) ሌᙋණჹ࡚‫ޤ܄‬೽ว‫ݹ‬ԴႵኳԄ‫ߥޑ‬ៈբҔϷխࣝፓ࿯ϐࣴ‫ز‬ ࣴ‫ز‬ғǺယৎՃ ࡰᏤ௲௤Ǻଯۘቺ. ௲௤. ύ୯ᙴᛰεᏢύ୯ᙴᏢࣴ‫܌ز‬. ࡚‫ޤ܄‬ཞ໾(Acute lung injury; ALI)ࢂ೷ԋ஻‫ޣ‬ԝΫ΋ঁख़ा‫ޑ‬ԋ ӢǶӵՖ‫ל‬ว‫ݯޑݹ‬ᕍ‫ׯࢂ܈‬๓ALI஻‫ޑޣ‬Ⴃࡕࢂख़ा‫ޑ‬ፐᚒǴρԖ೚ ӭЎ᝘ࡰрύᙴᛰӧჹ‫ޤל‬೽࡚‫܄‬ว‫ݹ‬Ԗ‫ځ‬ԋਏǴԶሌᙋණߡࢂ‫ځ‬ύჹ ‫ޤל‬೽࡚‫܄‬ว‫ݹ‬ख़ा‫ޑ‬БᏊǴՠ‫לځ‬ว‫ޑݹ‬ᐒ‫ڋ‬ϝฅόమཱǶ ࣁΑࣴ‫ز‬ሌᙋණჹ‫ޤל‬೽࡚‫܄‬ว‫ޑݹ‬ᐒ‫ڋ‬Ǵ‫ॺך‬а਻ᆅ‫׫‬ᆶLPS ᇨวԴႵ‫ޤ‬೽࡚‫܄‬ว‫ݹ‬Ǵӕਔᗯ१ሌᙋණǴ่݀ว߄όፕӧ‫ޤ‬೽ว‫ޑݹ‬ ੰ౛ಔᙃ΢ǵύ‫܄‬қՈౚ‫੆ޑ‬ዎኧໆǵ‫ޤ‬೽Нဍࡰ኱Ϸ΋਼ϯේϩ‫ޑݜ‬ ໆ΢Ǵࣣё‫ډـ‬ሌᙋණԖ෧጗‫ޤ‬೽ว‫ޑݹ‬ԋਏǶ‫ॺך‬аELISA Ϸ RT-PCR ٰୀෳӚᅿว‫࣬ݹ‬ᜢ‫ޑ‬ಒझᐟનϷᖿϯӢηǴӵTNFĮǵIL-1ȕǵ IL-6ǵKCǵMCP-1ϷMIP-2ǴሌᙋණࣣԖᓸ‫ޑڋ‬բҔǴӕਔ‫ॺך‬Ψว౜ ሌᙋණёᇨวεໆ‫לޑ‬ว‫ݹ‬ಒझᐟનIL-10‫ޑ‬ϩ‫ݜ‬Ƕ‫ॺך‬ճҔႵѮᏘಒझ RAW 264.7ଞჹTNFĮϷIL-10 ຾ՉᡏѦࣴ‫ز‬Ƕሌᙋණǵሌ޸ǵೱᙋϷФ ⣢ჹLPSᇨวౢғTNF-ĮԖܴᡉᓸ‫׭‬բҔǴ૘޺ǵᖓ಻Ϸ఩ԮယϿ೚ᓸ ‫ڋ‬բҔǶԶሌᙋණϷߎሌ޸ёаቚуLPSᇨวϩ‫ݜ‬IL-10Ƕ җሌᙋණ‫ޑ‬HPLCԋҽϩ‫݋‬ύǴ‫ډפ‬ሌᙋණࡰ኱ԋϩᆘচለǵЕ ช૛નǵФ⣢નϷᐈҜનǴ‫ځ‬ύനεໆ‫ޑ‬ԋҽࣁᆘচለǴ‫ॺך‬Ψ᛾ჴᆘ চለ཮ቚуLPSᇨวRAW 264.7ϩ‫ݜ‬IL-10ǹќѦ‫ॺך‬Ψว౜ᐈҜનё‫׭‬ ‫ڋ‬A549ϷTHP-1‫ޑ‬TNFĮǵIL-1ȕǵIL-6ǵIL-8‫ޑ‬ϩ‫ݜ‬Ǵӕਔ่݀ΨᡉҢ ᐈҜનё‫ڋ׭‬ว‫ݹ‬ಒझ‫ߕޑ‬๱բҔϷICAM-1ᆶVCAM-1‫߄ޑ‬౜Ƕ‫ॺך‬຾ ΋‫؁‬ว౜ሌᙋණᐈҜનё෧ϿNF-țBᆶAP-1ჹ‫ځ‬௓‫ׇڋ‬ӈ่ӝբҔǴ٠ ЪᐈҜનёߔᘐIțB ϷMAPK ၡ৩ǶӢԜǴሌᙋණԖ‫ڋ׭‬LPS೷ԋ‫ޤޑ‬ ೽ว‫ݹ‬բҔϷխࣝፓ࿯բҔǴԖշ‫ޤ܄࡚ܭ‬೽ว‫ݯޑݹ‬ᕍǶ ᜢᗖӷǺሌᙋණ,ᐈҜન,ᘞқණ,ᆘচለ,ϣࢥન,ಒझᐟન,ᙯᒵӢη,ߕ๱ Ӣη x.

(11) ಃ΋ക. ߻ ‫ق‬. ࡚‫ޤ܄‬ཞ໾(Acute lung injury; ALI)ࢂ೷ԋ஻‫ޣ‬ԝΫ΋ঁख़ा‫ޑ‬ԋ ӢǶӧᖏ‫׉‬΢‫୷ک‬ᘵࣽᏢ΢ჹALI‫ੰޑ‬౛ғԋ΢ςԖࡐӭ‫ޑ‬ΑှǶਥᏵ ೭٤ࣴ‫่݀ز‬Ǵς࿶ஒࣴ‫ޑز‬Бӛᙯ‫ډ‬ΑӵՖ‫ל‬ว‫ݯޑݹ‬ᕍ‫ׯࢂ܈‬๓ ALI஻‫ޑޣ‬Ⴃࡕ΢Ǵҁࣴ‫ز‬ஒ௖૸ύᙴᛰӧჹ‫ל‬ALI‫ޑ‬ԋਏϷ‫לځ‬ว‫ޑݹ‬ ᐒ‫ڋ‬Ƕ ԾAshbaugh1฻Γ‫ ܭ‬1967 ԃว߄Α࡚‫֎ڥ܄‬૰ᆃ‫ٯੰޑ‬Ǵ஻‫ޣ‬Ӣ Ѧ໾‫܈‬གࢉࡕр౜࡚‫֚֎ڥ܄‬ᜤǴեՈ਼Ǵ‫ޤ‬೽ᔈ‫܄‬෧եǴૅ೽XӀԖ ‫ୁٿ‬᝕ᅐ‫੆܄‬ዎǴа໺಍‫ݯ֎ڥ‬ᕍБ‫ݤ‬ค‫ݤ‬ᕖ҅‫ځ‬եՈ਼฻౜ຝǶੰ౛ ΢р౜‫ޤ‬ಔᙃܴᡉ‫ޑ‬кՈǴ‫ݰޤ‬Ϸ໔፦НဍǴ‫ݰޤ‬༥ഐǴ‫܄ݹ‬ಒझ੆ዎǴ ‫ݰޤ‬ϣԖ೸ܴጢ(hyaline membrane)ϐ‫׎‬ԋϷᠼᆢϯੰᡂǶҗ‫ܭ‬Ԝੰ߈ ՟ཥғ‫ٽ‬ϐ‫ึ֎ڥ‬ॐੱংဂǴAshbaughᆶPetty2‫ ܭ‬1971 ԃஒϐ‫ڮ‬Ӝࣁ ԋΓ‫ึ֎ڥ‬ॐੱংဂǴ(Adult respiratory distress syndromeǴARDS)Ƕ ࿶җᙴࣚ‫ޑ‬ոΚǴჹ‫ܭ‬Ԝੰቶ‫زࣴޑݱ‬Ǵว౜‫ޤ‬೽‫ޔޑ‬ௗ໾্‫ޤ܈‬Ѧ‫ޑ‬ ‫ځ‬Ѭ੯஻Ǵ֡ёᏤठԜ΋ੱংဂ‫ޑ‬วғǶ ԶԜᅿ࡚‫ޤ܄‬ཞ໾ࢂԖᇸख़ϐϩǴARDS҅ࢂԜੰำനᝄख़‫߄ޑ‬ ౜Ƕᖏ‫׉‬΢ᏤठARDS‫ޑ‬চӢǴёϩࣁ‫ޔ‬ௗ‫ޤ‬໾্‫ک‬໔ௗ‫ޤ‬໾্‫ٿ‬ᅿǶ ໔ௗ‫ޤ‬໾্Ǵࢂҗ‫ޤܭ‬аѦ‫ޑ‬੯ੰ‫܈‬ཞ໾Ǵ࿶җӄ‫ي‬ว‫ݹ‬ϸᔈ‫ޑ‬ᐟࢲǴ ٬Ոύಒझᐟન‫܈‬ಒझϩ‫ޑݜ‬ว‫࣬ݹ‬ᜢϟ፦ᐚࡋቚуǴԶ໔ௗ೷ԋ‫ޤ‬ཞ ໾Ƕ೷ԋARDS‫ޑ‬ᖏ‫׉‬੯ੰԖ‫ޔ‬ௗ‫ޤ‬໾্ǵ֎Εगనǵᝄख़ૅᄂѦ໾ǵ ‫ޤ‬਋໾ǵ᝕ᅐ‫ޤ܄‬གࢉǵಒ๵ǵੰࢥǵ‫៶ޤ‬ᙝǵᝄख़௳Ոੱǵ࡚‫܄‬ી᠌ ‫ݹ‬ǵ‫ޤ‬౽෌ೌࡕ฻ǶҞ߻ᗨฅჹ‫ܭ‬ARDS‫ޑ‬ठੰᐒᙯၨԖΑှǴՠARDS ‫ݯޑ‬ᕍϝឦЍ࡭‫ݯ‬ᕍࣁЬǴԶ‫ځ‬ԝΫ౗ᗨԖΠफ़Ǵՠࢂϝฅᆢ࡭ӧ 40% Կ 60%‫ޑ‬ଯᗺǶ࡚‫ݹޤ܄‬നத‫ࢂޑـ‬ಒ๵གࢉǴЬाࢂӢࣁЬᡏϣӧٛ ᑇ‫ޑ‬ѨਏϷߏਔ໔٬Ҕ೯਻೛ഢ೷ԋǴԖε‫ܭ‬Οϩϐ΋‫ޑ‬٬Ҕ೯਻೛ഢ ೷ԋ೷ԋARDS3,4Ƕ ARDS஻‫ޣ‬ё‫ډـ‬ӧว‫ޤޑݹ‬೽ϣ‫ډـ‬εໆ‫ޑ‬ӭਡ ‫ޑ܄‬ύ‫܄‬ౚ5,6Ϸញ‫ܫ‬εໆ‫ޑ‬ว‫߻ݹ‬ϟ፦7,8Ǵ ೈқ፦ϩှሇનǴ਼ϩηϷ ΋਼ϯේϩη9.10Ƕ നࡕ೷ԋ‫ݰޤ‬Ꮫᆅ๚‫ډڙ‬ཞ্ǴԶᏤठεໆಔᙃ෧ 1.

(12) ϿӣࢬԶ‫ݰޤ‬ϣౢғΑ‫ޤ‬НဍϷ‫ׯ‬ᡂΑ‫ޑޤ‬೯਻ᐒ‫ڋ‬Ǵ่݀٬ள‫ޑޤ‬ඤ ਻ໆᝄख़‫ޑ‬લЮǴԶౢғΑARDS3,4,11ǶԾவARDS೏ว౜рٰǴЪஒᖏ ‫ुރੱޑ׉‬рǴՠᖏ‫زࣴޑ׉‬ᕴࢂόܰࣴ‫ز‬మཱ‫ځ‬ठੰ‫ޑ‬ᐒ‫ࣁڋ‬ՖǴߡ ว৖Α୏‫ނ‬ኳԄǴҗ୏‫ނ‬ኳԄё‫ډـ‬྽ੰΓวғARDSਔ‫ځ‬ᡏϣ‫ੰޑ‬౛ Ϸғ౛‫ޑ‬ᝄख़‫ޑ‬ᡂϯ΋΋և౜рٰΑǶ ύᙴᏢύ٠ค࡚‫ޤ܄‬೽གࢉੰӜǴՠਥᏵҁੰᖏ‫׉‬੝ᗺǴӭឦ‫ܭ‬ύ ᙴᏢ‫ޑ‬॥ྕ‫ޤ‬዗ຓǵഹ჌฻ጄᛑǶύᙴᏢᇡ㵝‫ݹޤ‬ഹ჌‫ੰޑ‬ӢхࡴϣӢ ‫ک‬ѦӢ‫ঁٿ‬Бय़ǶѦӢࢂѦག॥ٕ(॥ൣ‫܈‬॥዗) ǴϣӢࢂ‫᠌ޤ‬ብცǴ‫׎‬ ਻҂кǶՋᙴᏢᇡ㵝ࢂಒ๵‫ࢥੰک‬฻ੰচᡏགࢉ‫܌‬ठǴᆶ‫ي‬ᡏ‫לܢ‬Κ১ ΨԖᜢǶ‫ڀ‬ᡏឍញӵΠ:॥ٕȋੰচᡏǴхࡴಒ๵(ӵ‫ݹޤ‬ᚈౚ๵ǵߎ໳ Յဟ๻ౚ๵) ǵੰࢥ(ӵ‫֎ڥ‬ၰӝझੰࢥǵဏੰࢥ) ǵЍচᡏ(ӵ‫ݹޤ‬Ѝচ ᡏ) ǵচᙝ(ӵьМ‫៶ޤ‬ᙝ) ฻Ƕՠȸੰচᡏȹόૈֹӄ฻ӕ‫ܭ‬ȸ॥ٕȹ ‫ۺཷޑ‬ǶύᙴᏢவֻᢀ‫ޑ‬᏾ᡏ‫ࡋفޑ‬ᇡ᛽ੰӢǴᇡ㵝੯ੰ‫ޑ‬วғࢂҗ‫ܭ‬ Γᆶᕉნ‫ک‬ΓԾ‫ޑي‬ᛙᄊ೏ઇᚯ‫܌‬ठǴ‫ݙ‬ख़ჹ೭ᅿѨᑽ‫ރ‬ᄊ‫ޑ‬ፓ࿯ǴԶ όు‫ز‬Ꮴठ೭ᅿ‫ރ‬ᄊ‫ঁޑ‬ձӢનǶύᙴᏢ‫ੰޑ‬Ӣࢂ֖Ԗੰᐒཀက‫ޑ‬ᆕӝ ཷ‫ۺ‬ǴႽȸ॥ٕȹ೭ᅿᙁዅ‫߄ޑ‬ॊǴ੮๏аՋᙴᏢ‫ޑ‬БԄࣴ‫܌ز‬ளѐ༤ к‫ޜޑ‬໔ࢂคज़‫ޑ‬ǶύᙴᏢჹ‫ݹޤ‬ഹ჌ੰ౛ᐒ‫ޑڋ‬ፕॊࢂ:࿀ߔ਻ၰǴ‫ޤ‬ ਻ഈᢠǴ‫ޤ‬Ѩ࠹๘Ƕᜢᗖࢂ‫ޤ‬ഈǴ่݀ࢂ࠹๘ѨதǶ೭΋ᐒ‫ڋ‬ёаҔ౜ жᙴᏢᇟ‫ق‬ዴϪӦှញ:Ѝ਻ᆅℚጢ‫ݰޤک‬ᏛкՈНဍǴЍ਻ᆅ‫ݰޤک‬ϣ ϩ‫ނݜ‬ቚӭǴᏤठ೯਻‫ک‬ඤ਻фૈምᛖ(࿀ߔ਻ၰǵ‫਻ޤ‬ഈᢠ) ǴЇଆલ ਼‫ک‬Β਼ϯᅹ‫ޑ‬ゅ੮(࠹๘Ѩத)Ƕ‫ݹޤ‬ഹ჌‫ޑ‬த‫ـ‬ᒣ᛾Ј໚຀૰‫ੰޑ‬ᐒ ΨёҔЈΚ૰ᆃ‫ޑ‬㰗ғှញ12Ƕ ӧ೚ӭठੰྍύ‫ޑ‬ӭᗐᡏȐLipopoloysaccchrideǴLPSȑЬाٰԾ ਱ើМ഍‫܄‬ಒ๵(gram-negative bacteria)ಒझᏛѦቫિӭᗐᡏ,཮٬‫ޤ‬ ‫ݰ‬ϣౢғΑ‫ޤ‬НဍϷ‫ׯ‬ᡂΑ‫ޑޤ‬೯਻ᐒ‫ڋ‬Ǵ่݀٬ள‫ޤ‬ඤ਻ໆᝄख़‫ޑ‬લ ЮǴԶౢғΑARDS3,4Ƕӧ೚ӭठੰྍύLPSࢂቶ‫ݱ‬೏ௗ‫ڙ‬೷ԋ‫ޤ‬೽࡚ ‫܄‬ว‫ݹ‬ന٫‫ޑ‬ᇨว‫ނ‬ǴΨࢂനௗ߈ᖏ‫׉‬΢࡚‫܄‬ว‫ޑݹ‬ኳԄ13,14Ƕ‫܌‬аϞ В೚ӭЎ᝘аϣࢥનٰᇨว࡚‫ޤ܄‬೽‫ੱݹ‬ϸᔈаբ຾΋‫زࣴ؁‬Ƕ ሌᙋණྍ‫ܭ‬మжᙴৎֆᗫ೯‫܌‬๱Ƞྕੰచᒣȡϐύ‫૶܌‬ၩǴࢂ֖΋ ‫ۓ‬К‫ ޑٯ‬10 ᅿύᛰಔԋǴӧѠ᡼ǴሌᙋණࢂԖ‫ޑۓڰ‬К‫ٯ‬Ϸ኱ྗᇙԋ 2.

(13) ‫ޑ‬ύᛰೀБǴӧᖏ‫׉‬΢Ϸ໺಍ᙴᏢ΢Ǵሌᙋණ࿶த೏٬Ҕӧ‫֎ڥ‬ၰ‫ޑ‬੯ ੰǴӵགߵǵ਻ᆅགࢉǵϷ‫ݹޤ‬฻ǶࣁΑࣴ‫ز‬ሌᙋණ‫ޑ‬բҔǴ‫ॺך‬ᒧҔ җ਻ᆅឲΕ LPS ‫ޑ‬БԄǴࡌҥԴႵ‫ޤ܄࡚ޑ‬೽ว‫ࣁݹ‬ኳԄǴ຾Չ‫ݯ‬ᕍǴ ӕਔճҔ HPLC ჹሌᙋණ৖໒ϩ‫݋‬Ǵ຾΋‫ډפ؁‬ЬाԋҽǴӆаႵѮᏘ ಒझ RAW 264.7 ຾Չ LPS ‫װ‬ᔐሌᙋණ٠а‫ځ‬Ьाԋҽ‫ݯ‬ᕍǶ཮ᢀჸႵ ‫ޤ‬೽ख़ໆǵ‫ࢱؑݰޤ‬నᕴಒझኧǵೈқ፦ᐚࡋǵ΋਼ϯේ‫߄ޑ‬౜ϷӚᅿ ಒझᐟનǴӵ TNFĮǴ IL-1ȕǴ IL-6Ǵ KCǴ MIP-2Ǵ MCP-1Ǵ IL-4Ǵ IL-10 and IL-12 ฻ϐᡂϯǴ٠ჹ‫ޤ‬ಔᙃ຾Չ RT-PCR ϩ‫݋‬ϷਡύᙯᒵӢ ηȐNF-kB and AP-1ȑа EMSA ϩ‫݋‬ǶჹႵ‫ޑ‬ѮᏘಒझ RAW 264.7 Ψ ཮຾ՉಒझᐟનϷਡύᙯᒵӢηȐNF-kB and AP-1ȑEMSA ϩ‫݋‬Ƕ‫ॺך‬ གྷຑ՗ሌᙋණჹ LPS ೷ԋ‫ޤޑ‬೽ว‫ݹ‬ǵխࣝፓ࿯բҔϷᐒᙯǴ٠Ъϩ‫݋‬ ‫ځ‬ύЬाԋϩӧሌᙋණύ‫ޑ‬ӦՏϷᐒᙯǶ. 3.

(14) ಃΒക. Ў᝘௖૸. Ȑ΋ȑύᙴЎ᝘ӣ៝ ಃ΋࿯ʳ ‫ݹޤ‬ύᙴੰӜ‫ޑ‬ౢғᆶ‫ݮ‬ॠ ύᙴᏢ٠ค࡚‫ޤ܄‬೽གࢉੰӜǴՠਥᏵҁੰᖏ‫׉‬੝ᗺǴӭ ឦ‫ܭ‬ύᙴᏢ‫ޑ‬॥ྕ‫ޤ‬዗ຓǵഹ჌฻ጄᛑǶ ಃΒ࿯ʳ ‫ੰݹޤ‬Ӣੰᐒ ࡚‫ޤ܄‬೽གࢉࢂᖏ‫׉‬த‫ੰـ‬ǵӭวੰǶᖏ‫׉‬Ьा߄౜ࣁว ዗ǵࠏ჌ǵ਻ߦሷᅛǴᝄख़ਔࣗԿёр౜஭α‫ުܩ‬ǵ‫֚֎ڥ‬ᜤ ฻ੱ‫ރ‬ǶᅇǷ஭ҸඳȠ໾ൣፕȡύჹ॥ྕ൩ԖፕॊǶჹ‫ܭ‬ҁੰ ‫ੰޑ‬ӢੰᤵǴӭኧᙴৎᇡࣁҁੰठੰӢનԖϣѦ‫ٿ‬ӢǴϣӢࣁ ‫ޤ‬຀ፁѦό‫ڰ‬ǴѦӢࣁག‫ڙ‬॥ٕǴ॥ٕӭ਀ൣ਀዗ࣁੰǴ‫ځ‬ύ а॥዗ϐٕठੰӭ‫ـ‬Ƕ‫ۚޤ‬΢ขǴ໒ᘻ‫ܭ‬ሷǴѦӝҜЛǴЬ΋ ‫ي‬ϐ߄Ƕ॥ࣁԭੰϐߏǴΏࣁ໚ٕǴ‫܄ځ‬ᇸඦǴ‫ڀ‬Ԗϲණ‫ޑ‬੝ ᗺǶ‫܌‬а॥዗ϐٕߟ᠍ΓᥟǴӭவαሷԶΓǴӃҍ΢ข‫ޤ‬ፁԜ ϐᒏȸྕٕ΢‫ڙ‬Ǵ२Ӄҍ‫ޤ‬ȹǶ॥዗ҍ‫ޤ‬ǴѦ߾ፁ਻ॕߔǴҜ Л໒ӝόճǹϣ߾‫਻ޤ‬Ѩ࠹Ǵ๘फ़ѨᙍǶаठр౜ว዗ǵࠏ჌ ฻ຓংǶ‫ޤ‬Ь‫ݯ‬࿯Ǵ‫਻ޤ‬ॕഈǴ߾ՈՉόᄣǴр౜਻ᅉՈ྿ǹ ॥዗ϐٕΕ㚊Ǵ଍໺ЈᔼǴᇸ‫ޣ‬዗‫؟‬ᔼ഍Ǵᘋ໶Јઓǹख़‫ޣ‬዗ ഐЈхǴᆾጨЈᘻǴࣗԿ዗ΕᔼՈǴౢғ઻Ո୏Ո‫ޑ‬ᡂϯǶӢ Ԝҁੰӵ‫ݯ‬ᕍόள྽Ǵࡕයёр౜Ј໚຀૰‫ٕ܈‬ഐല഍Ƕ౜ж ࣴ‫ز‬᛾ჴǴҁੰၸำύёр౜ܴᡉ‫ޑ‬Ո྿᛾ংǶ. 4.

(15) ಃΟ࿯ʳ. ύᙴჹ‫ޑݹޤ‬ϩ᛾Ϸ‫ݯ‬ᕍཷፕ ύᙴᏢਥᏵ‫ݹޤ‬ഹ჌όӕ໘ࢤ߄౜р‫ރੱޑ‬ǵՁຝǵેຝϩ. 㵝:॥዗ҍ‫ޤ‬ǵ॥ൣഈ‫ޤ‬ǵ࿀዗ഈ‫ޤ‬ǵЈ໚຀૰ǵٕഐല഍฻ό ӕ᛾ࠠǶ ჹ‫ܭ‬ҁੰ‫ޑ‬ᒣຓ‫ݯ‬ᕍǴЬाϩ॥዗ҍ‫ޤ‬ǵ॥ൣഈ‫کޤ‬࿀዗ Ꮮ‫ޤ‬ΟࠠǶԖਔΨ཮р౜ᡂੱǴЬाԖЈ໚຀૰‫ک‬ϣഐല഍฻Ƕ ॥዗ҍ‫ݯࠠޤ‬аٌథ࠹‫ޤ‬ǵమ዗ϯ࿀Ǵаഞ‫׶‬ҡҒ෯ӝሌᙋණу ࣁЬǴᛰҔഞ໳ǵ‫׶‬Ϙǵғҡᆯǵሌ޸ǵೱᙋǵᖓ಻ǵਧఒǵФ ⣢ηǵҒ૛Ƕഹ৲࿀ሳ‫ޣ‬Ǵу♬㍟ηǵੈ‫҆ن‬ǹࠒോआဍੵภ‫ޣ‬Ǵ у৔υǵᙜՊǵЕጷጸǹว዗ଯ‫ޣ‬Ǵу໳᭷ǵεߙယǵਮचǵလ ਥǵНФ‫ف‬฻Ƕ ॥ൣഈ‫ݯࠠޤ‬аٌྕ໒‫ޤ‬ǵϯ࿀ЗࠏǴаΟ‫ܧ‬෯‫܈‬๮ᇂණу෧ࣁ ЬǴᛰҔഞ໳ǵ‫׶‬Ϙǵٛ॥ǵਧఒǵሼᡠǵҒ૛฻Ƕҗ‫ܭ‬ҁ᛾ܰ ‫ܭ‬ϯ዗‫܈‬ӭঋԖ዗ຝǴࡺத‫ܭ‬΢БύуΕమ዗ှࢥᛰ‫ނ‬Ƕҁࠠӵ ਀Ԗൣ໯Ǵаλߙᓪ෯‫ݯ‬ϐǶ ࿀዗Ꮮ‫ݯࠠޤ‬аమ዗࠹‫ޤ‬ǵᅗ࿀‫ۓ‬ഹǴБҔϖ߁෯ӝ♬㍟εවᘞ ‫ޤ‬෯у෧ࣁЬǴᛰҔഞ໳ǵ‫׶‬Ϙǵ♬㍟ηǵ᝵ηǵ໳᭷ǵ߁‫׺‬ǵ ߻चǵಒૡǵҒ૛Ƕ࿀ӭ‫ޣ‬ǴуϺ‫ޞ‬໳ǵᖌࠄࢃǹ࿀࿨‫ޣ‬ǴуԮ ᛅǴᱞჴǵఙηǵ‫҆ޕ‬ǵሌ޸ǵೱᙋǵε໳ǵ໳࢙ǵങည૛ǵҡ ᆯ฻Ǵӆଛӝ΋٤ϯ࿀Зࠏճ਻ϐࠔǴӵਧఒǵ‫҆ن‬ǵъহǵ๋ ⏎ǵීо޸ǵ‫׶‬ϘǵᱞෘǵഋҜǵଭେႍ฻ǶΨԖаሌᙋණǵί ߎဘಳ෯ǵഞ‫׶‬ҡҒ෯฻ࣁЬ‫ޣ‬Ǵӆ፾྽ଛаమ‫ޤ‬ϯ࿀ϐࠔǶᒿ ๱ჹҁੰ੝౦‫ੰ܄‬চᡏ‫ޤک‬೽‫ੰޑੱݹ‬౛ᡂϯ‫ޑ‬ుΕᇡ᛽Ǵதᒧ Ҕమ዗ှࢥǵ‫ל‬๵੃‫ݹ‬բҔၨம‫ޑ‬ύᛰǴӵങည૛ǵ߁‫׺‬ǵъ݄ ጪǵ໳᭷ǵሌ޸ǵߎⶊഝਥǵқ޸ೂՁ૛฻ಔБ‫ݯ‬ᕍǶӧ‫ੰݹޤ‬ ౛ၸำύ۳۳р౜ၨᝄख़‫ޑ‬Ո྿ຓংǴӢԜǴӧ‫ݯ‬ᕍၸำύ‫ډـ‬ ԖՈ྿ຓংਔǴ೯தуҔ΋٤ࢲՈϯ྿ϐࠔǴࡽගଯΑᕍਏǴΞ ё෧ϿΑᡂੱ‫ޑ‬วғǶ. 5.

(16) ಃѤ࿯ ሌᙋණ‫ޑ‬Ў᝘ӣ៝ ሌᙋණࢂ‫ך‬୯మжᙴৎֆᅨ(ӷᗫ೯) ബ‫ޑڋ‬๱ӜύᛰБ ᏊǴೀБҗ10‫ښ‬ύᛰಔԋǶ၀Б‫ܭـۈ‬మж1788 ԃ‫څ‬ӑр‫ޑހ‬ ֆᅨ‫ޑ‬ж߄ύᙴ๱բȠྕੰచᒣȡǴ‫ڀ‬Ԗٌథှ߄ǵమ዗ှࢥ‫ޑ‬ фਏǴЬ‫ݯ‬॥ྕ߃ଆǴ྽ਔа‫ݯ‬ᕍ॥ྕǵྕ዗ੰаϷࢌ٤ᚇੰឦ ‫ٕܭ‬ӧፁϩǵ΢ข‫ޣ‬ԶᆪӜǴ٠΋‫ݮޔ‬ҔԿϞǶ ᛰ‫ނ‬ಔԋ: ೱᙋǴሌ޸ǴधਧఒǴᖓ಻Ǵ఩ԮယǴғҒ૛Ǵ ૘޺ᕞǴ఩‫ل‬೟ǴФ⣢ηǴဘਥǶ Ҕ‫ݤ‬Ҕໆ: ᗲဘਥ෯ྞǴ३਻εрǴջ‫ܺڗ‬ǴϮၸฆǶੰख़‫ޣ‬ऊ Βਔ΋ܺǴВΟܺǴ‫ڹ‬΋ܺǹᇸ‫ޣ‬Οਔ΋ܺǴВΒܺǴ‫ڹ‬΋ܺǴ ੰόှ‫ޣ‬բӆܺǶ Ь‫ݯ‬: ྕੰ߃ଆǴว዗คԠǴ‫܈‬ԖԠόᄣǴ༾ൾൣǴᓐภα ෰Ǵࠏ჌ࠒภǴՁӾआǴऱᖓқ‫܈‬ᖓ໳Ǵેੌኧ‫ޣ‬Ƕ ౜жᖏ‫׉‬΢Ҕ‫ݯܭ‬ᕍࢬՉ‫܄‬གߵǵ࡚‫ࡧ܄‬ਲဏ‫ݹ‬ǵࠒോ‫ݹ‬ǵ ‫ݹޤ‬ǵ㫶੶ǵഞ੶ǵࢬՉ‫܄‬ဋဏ‫ݹ‬฻ੰࢥ‫܄‬གࢉ੯ੰǴࢂύᙴᖏ ‫׉‬΢ቶ‫ݱ‬٬Ҕ٠ЪᕍਏዴϪ‫ޑ‬ύᛰБᏊϐ΋18Ƕ ߈ԃٰǴԖ೚ӭΓ௦Ҕ౜жᛰ౛Ꮲࣴ‫ز‬Б‫ݤ‬ჹሌᙋණ຾Չు Ε‫زࣴޑ‬Ǵ่݀և౜Ǵሌᙋණ‫ڀ‬Ԗှ዗ǵᙼภǵ‫ל‬๵ǵ‫ࢥੰל‬ǵ ှࢥǵ‫ݹל‬ǵ‫ל‬ၸ௵฻բҔǶ‫ॺך‬ଞჹ߈ԃٰჹሌᙋණӧ΢‫֎ڥ‬ ၰགࢉϷ‫ޑݹޤ‬ᖏ‫׉‬ᔈҔᆶᛰ౛Ꮲჴᡍࣴ‫ز‬բ᏾౛Ƕ ࡚‫܄‬΢‫֎ڥ‬ၰགࢉǴ㵝ᖏ‫׉‬΢த‫ੰـ‬ӭวੰǶҁੰឦύᙴྕ ੰȹ॥ྕȹ‫ޑ‬ጄᛑǶሌᙋණբ㵝‫ݯ‬ᕍ॥ྕٕ዗ӧ‫ޤ‬ፁ‫ޑ‬ЬБǴ၀ Бϝฅᗋࢂ౜ж‫ݯ‬ᕍ΢‫֎ڥ‬ၰགࢉ‫ޑ‬ЬाБᏊǶ२Ӄࢂֈܴඁ௖ ૸ගଯаύᙴᛰࣁЬ‫ݯ‬ᕍ࡚‫܄‬΢‫֎ڥ‬ၰགࢉ‫ޑ‬ᕍਏǶҔԾᔕу‫ښ‬ ሌᙋණࣁЬБ‫ݯ‬ᕍ࡚‫܄‬΢‫֎ڥ‬ၰགࢉ62‫ٯ‬Ǵ٠ᆶ٬ҔՋᛰ‫ݯ‬ᕍ ಔ50‫ٯ‬բКၨᢀჸǴ่݀у‫ښ‬ሌᙋණ‫ݯ‬ᕍ࡚‫܄‬΢‫֎ڥ‬ၰགࢉᆶ ൂપ٬ҔՋᛰԖܴᡉ‫ޑ‬ৡ౦19Ƕ ቅᖚஒѦགଯ዗ੱ஻‫ٽ‬ϩ㵝‫ٿ‬ಔǴҘಔҔᚈ໳ೱણଞᓉેᗺ ᅀӝሌᙋ೓‫ݰ‬Ꮚ‫ݯ‬ᕍǴΌಔҔߙᡖનᓉેᗺᅀǶ‫ݯ‬ᕍ่݀ᡉҢǴ 6.

(17) ҘಔᕴԖਏ౗㵝95. 34 % ǴΌಔ㵝90. 6 %ǶҘಔӧଏ዗Бय़ܴ ᡉᓬ‫ܭ‬Όಔ( P < 0. 05) 20Ƕ 1998ԃЦ‫ד‬ཥஒ275‫ٯ‬གߵੰΓϩԋҘΌ‫ٿ‬ಔǴҘಔ๏ϒа ሌᙋණǵ݈ᙔਥǵIbuprofen฻ᇙԋ‫ޑ‬ቚਏሌᙋТ‫ݯ‬ᕍǴΌಔϒ மΚሌᙋТ‫ݯ‬ᕍǶ3 Ϻϣ‫ݯޑ‬ᘰ౗ǵ୷ҁ‫ݯ‬ᘰ౗ǵᕴԖਏ౗ϩ ձ㵝ǴҘಔ61. 45 %ǵ23. 64 %ǵ96. 36 %ǹΌಔ20.69 %ǵ10. 34 %ǵ87. 36 %21Ƕ ೭٤ჴᡍ่݀߄ܴǴሌᙋණൂᐱ٬ҔǴ‫ݯ‬ᕍ΢‫֎ڥ‬ၰགࢉԖ ᡉ๱‫ޑ‬ᕍਏǴՠࢂऩᆶϯᏢᛰ‫ނ‬ᛰ‫ނ‬ᖄӝ٬ҔǴ߾‫ݯ‬ᕍਏ݀ाܴ ᡉᓬ‫ൂܭ‬Ҕሌᙋණ‫ޣ܈‬ϯᏢᛰ‫ނ‬Ǵᇥܴࢌ٤ϯᏢᛰ‫ނ‬ᆶሌᙋණ࣬ ϕ໔ԖቚਏբҔǶ ‫֎ڥ‬ၰགࢉЬाҗੰࢥЇଆ‫ޑ‬Ǵܰҗ΢‫֎ڥ‬ၰགࢉᝩว㵝਻ ᆅ‫ݹޤکݹ‬฻Π‫֎ڥ‬ၰགࢉǶ၀ੰЬाวੰੱ‫ރ‬㵝ว዗ǵࠏ჌ǵ ૅ඀ǵα෰ǵՁआऱ໳ǵેኧǴឦ‫ܭ‬ύᙴ‫ޤ‬዗ੰ‫ੰྕک‬ጄᛑǶ ઽλ૜ϷቅӀϼ௦ҔሌᙋණྞᏊǴ٠аੰࢥᶉу‫ࢥੰל‬αܺ నբჹྣǴ‫ݯ‬ᕍ102 ‫ٯ‬Π‫֎ڥ‬ၰགࢉ஻‫ޣ‬Ǵ่݀߄ܴǴ‫ݯ‬ᕍಔ ‫ޑ‬ᡉਏ౗‫ک‬ᕴԖਏ౗ᓬ‫ܭ‬ჹྣಔ( P < 0. 05) 22Ƕ ӧሌᙋණྞᏊу෧‫ݯ‬ᕍλ‫ݹޤٽ‬Бय़Ǵचۚ৲ว౜Ǵሌᙋණ Ҕ‫ܭ‬ᖏ‫ݯ׉‬ᕍჺҔ‫ל‬๵નԶᕍਏόᡉ๱‫ݹޤޑ‬஻‫ޣ‬Ԗ‫܌‬շ੻ 23. Ƕ‫ص‬లถ฻೯аሌᙋණ‫܈‬қ߁෯у෧Ǵੱख़‫ޣ‬уҔ‫ל‬ғનǴᕍ. ਏᓬ‫ൂܭ‬Ҕ‫ל‬ғન‫ݯ‬ᕍ24ǶॡߪҔሌᙋණଛӝߙᡖન٬ҔǶวύ Ջᛰ‫ݯ‬ᕍಔᕍਏၨൂҔՋᛰಔ‫ז‬25Ƕ ӧԴΓ‫ݹޤ‬Бय़Ǵླྀ‫ྷڬ‬ଞჹ60‫ٯ‬Դԃ‫ݹޤ܄‬٬Ҕሌᙋණ ӝ‫ံٳ‬ύ੻਻෯‫ݯ‬ᕍ‫܈‬ӝ‫ٳ‬ਬ๸໯‫ݯ‬ᕍКၨǴ‫ٿ‬ಔԖਏ౗ࣣ 50%ǴԶӝ‫ံٳ‬ύ੻਻෯ಔԋਏၨᓬ26Ƕ ᖏ‫׉‬ၗ਑ٰ࣮ǴሌᙋණჹӚԃសቫ‫ݹޤޑ‬Ǵόፕӧൂᐱу෧ Б٬Ҕ‫܈‬ଛӝՋᛰ٬ҔǴ೿ૈܴᡉ‫ׯ‬๓‫ރੱޑݹޤ‬ϷᕭอੰำǶ ӧჴᡍ࠻㚊‫ޑ‬ᛰ౛ࣴ‫ز‬΢ΨԖ࣬྽ӭ‫ޑ‬ԋ݀ว߄Ǵёெ‫ޑ‬ ࢂǴว߄ӧ୯ሞSCI‫ޑ‬ፕЎόӭǴՠ‫ॺך‬൩ჴᡍٰ่࣮݀Ǵόᜤ ࣮р‫่݀ځ‬ёᆶҁፕЎ‫่࣬݀ޑ‬ϕ՘᛾Ƕ 7.

(18) ሌᙋණှ዗Бय़Ǵ೚ӭ୏‫ނ‬ჴᡍ߄ܴǴሌᙋණჹόӕठ዗Ꮚ ‫܌‬Їଆ‫ޑ‬ৎ‫ټ‬ว዗֡Ԗܴᡉ‫ޑ‬ှ዗բҔǹ‫׹‬Κै฻ว౜ሌᙋණૈ ୼ှନठ዗চჹWistar εႵྕࡋ௵གઓ࿶ϡ‫ޑ‬բҔǴ᛾ܴ၀ᛰ 㵝ύኰ‫܄‬ှ዗ᛰǴЪ‫ځ‬բҔচ౛όӄӕ‫ܭ‬ှ዗ᙼภᜪᛰ‫ނ‬30Ƕ‫׹‬ М฻೯ၸа໾ൣୋ໾ൣҘΌషӝ๵भբ㵝୏‫ނ‬ᢀჸሌᙋණჹᡏ Ѧ୻ᎦᚆൂਡಒझȐMNCȑӝԋញ‫ܫ‬ϣғठ዗চȐendogenous pyrogenǴEPȑǴ ‫ޑ‬ቹៜǴ่݀߄ܴ‫ځ‬ჹEP ӝԋคܴᡉቹៜǴ ගҢሌᙋණ‫ޑ‬ှ዗բҔёૈӧ‫ߔܭ‬ᘐEP 㰗ғаࡕ‫ޑ‬ᕉ࿯31Ƕ ൤ܼ‫ػ‬฻ΓΨჹሌᙋණӧεқႵϐᛰ‫୏ނ‬ΚᏢୖኧ຾Չࣴ ‫ز‬Ƕ่݀߄ܴሌᙋණ‫ޑ‬നλଆਏᏊໆ㵝0. 18 g/ kg ǴբҔය㵝 6. 4 λਔǴᡏϣғ‫࣬ނ‬྽ᛰໆ‫੃ޑ‬ନъ૰ය㵝1. 11λਔǶќѦ аวԠ‫ޑ‬ᛰਏჴᡍ‫ݤ‬᛾ܴሌᙋණૈߦ຾εႵ‫ޑ‬Ԡဏϩ‫ݜ‬Ǵሌᙋණ ‫ޑ‬നեଆਏᏊໆ㵝0. 267 g/ kgǴ࣬྽‫ܭ‬ᖏ‫׉‬฻ਏᏊໆ‫ޑ‬ਏᔈ੃ ଏъ૰ය㵝3. 90λਔǴਏᔈᆢ࡭ਔ໔㵝23. 71λਔǴਏᔈၲঢ়ਔ ໔㵝2. 21λਔ33,34Ƕ ሌᙋණӕਔΨ೏᛾ܴ‫ڀ‬Ԗࡐம‫ݹלޑ‬ᆶ‫ל‬ၸ௵բҔǴૈቚம ѮᏘಒझჹ౦‫ޑނ‬տᏘૈΚϷ‫ڋ׭‬ಔᙃữ‫ޑ‬ϩ‫ݜ‬Ǵՠჹ5 -♏Յ ữ(5-HT)คܴᡉ‫ڋ׭‬Ǵჹ߻ӈဏનբҔΨၨ১35ǶΨว౜ሌᙋණ Ԗ‫׭‬๵բҔǴ୏‫ނ‬ೱុ๏ᛰѤϺࡕӆа‫ݹޤ‬ౚ๵གࢉǴ߾ёܴᡉ ෧ϿԝΫ౗ǹሌᙋණёගଯ୏‫לޑނ‬གࢉΚǶᡏѦΨჹ‫ݹޤ‬ౚ๵ Ԗ‫׭‬๵բҔ36Ƕ ӧ୯ሞයтё‫ـ‬ሌᙋණ‫زࣴޑ‬ύǴВҁϷύ୯εഌ֡ว߄ሌ ᙋණ‫ڀ‬Ԗ‫ל‬གࢉϷ‫ޑࢥੰל‬բҔ15,16Ƕ ӕਔ‫פ‬рሌᙋණύ‫ٿ‬ᅿ ύᛰɡҒ૛ϷФ⣢ηǴԖ‫ޑࢥੰל‬բҔ15Ǵ Kurokawa Ψගр ሌᙋණӧ‫ל‬གࢉ‫ޑ‬ၸำύՈమύ‫ޑ‬serum IL-1Į཮೏ගܹǴٰჹ ‫ל‬གߵੰࢥ16Ƕ җՅቫϩ‫ٰ݋‬ϩ‫݋‬ሌᙋණ‫ޑ‬ԋϩρ‫ډפ‬Α19ᅿ ϯᏢ‫ނ‬፦17Ǵӵ arctiinǴ phillyrinǴ forsythiasideǴ liquiritigeninǴ liquiritinǴ genisteinǴ formononetinǴ daidzeinǴ glycitrinǴ 3,3'Ǵ 4-tri-omethlellagic acid Ǵchlorogenic acidǴlinarinǴ acacetinǴ hesperidinǴ isoquiritigeninǴ isoquiritin and hyperoside17ǶΨ 8.

(19) Ԗ่ӝᛰਏᛰ౛բҔჹಔԋፄБ‫ښൂޑ‬ᛰ‫ޑ‬Ԗਏԋϩ຾Չࣴ ‫ز‬Ǵӵว౜ߎሌ޸ύ‫ޑ‬ᆘচለ(chlorogenic acid)ǵ౦ᆘচለ (Isochlorogenic acid)‫ڀ‬Ԗ‫ࢥੰל‬ǵ‫ל‬๵‫ޑ‬բҔǹೱᙋύ‫ޑ‬ೱᙋ ✊㧿(forsythiaside)‫ڀ‬Ԗ‫ל‬๵բҔ฻36 ǶᗨฅԖ೚ӭჹሌᙋණ‫ޑ‬ ࣴ‫่݀ز‬Ǵՠࢂሌᙋණჹᡏϣ‫ޑ‬խࣝፓ࿯‫ޑ‬ᐒ‫ڋ‬Ǵϝۘ҂‫פ‬рǴ ҁჴᡍճҔLPS‫ޤ‬೽གࢉႵኳԄ຾Չሌᙋණჹ‫ޤ‬೽ϣ‫ޑ‬խࣝፓ ௓ᐒ‫ڋ‬ǴΨჹ΢ॊ‫زࣴޑ‬ԋ่݀ගрӝ౛‫ޑ‬ᐒ‫ڋ‬Ƕ (Β) ౜жᙴᏢЎ᝘ӣ៝ ಃ΋࿯ ϣࢥનϷ‫ځ‬ౢғ‫ޑ‬ว‫ݹ‬ϸᔈ ϣࢥનЬाٰԾ਱ើМ഍‫܄‬ಒ๵‫ޑ‬ಒझᏛѦቫિӭᗐᡏ (LPS)Ǵ‫่ځ‬ᄬЬाх֖‫ٿ‬೽ϩǺӭᗐᡏ(polysaccharide)Ϸિ፦ (lipid)ǴԶ೷ԋғ‫ނ‬ᡏࢥ‫܄‬ϸᔈЬाٰԾિ፦‫ޑ‬೽ϩǴΞᆀࣁlipid AǶϣࢥનӧғ‫ނ‬ᡏϣ཮ࢲϯѮᏘಒझϷ‫ځ‬д࣬ᜢ‫ޑ‬ว‫ݹ‬ಒझǴ ྽LPSΕߟғ‫ނ‬ᡏϣਔѸ໪ӃᆶՈዀௗ‫ڙ‬ᡏ่ӝࡕǴߡ཮‫ڈ‬ᐟѮ Ꮨಒझញ‫ܫ‬΋٤‫ނ‬፦Ǵӵಒझᐟન(cytokines)Ƕ྽ѮᏘಒझ‫ڙ‬ LPS‫ڈ‬ᐟϩ‫ݜ‬፾ໆ‫ޑ‬ፓ࿯‫ނ‬፦ਔǴჹ‫ي‬ᡏёаౢғߥៈբҔǴ຾ Զၲ‫ډ‬ఠ๵բҔǶฅϣҜಒझញ‫ܫ‬рԾҗ୷ǵೈқሇનԶઇᚯಔ ᙃǴ‫׎‬೷ಔᙃ೯೸‫܄‬ቚуǶᕉნύLPSቶ‫ݱ‬Ӹӧ‫਻ޜܭ‬ԦࢉǵԖ ᐒણლϷྟᜦύǴёᙖҗ‫਻ޜ‬໺ኞǴߏයኪ៛ϣࢥનё೷ԋ೚ӭ ‫ޤ‬೽‫ੱݹ‬ϸᔈǴӵ਻ഹǵᄌ‫܄‬Ѝ਻ᆅ‫ݹ‬аϷ‫֎ڥ‬ၰߔ༞฻੯ੰǶ 2002 ԃǴVernooy฻Γஒλ໵ႵߏයаLPS਻ᆅϣ‫׫‬ᆶǴว౜ ߏයኪ៛LPSёаᇨว‫ޤ‬ಔᙃੰ౛‫܄‬ᡂϯǴӵ‫ޤ‬ᗹనಒझ౦ғ (mucus cell metaplasia)ǵ‫ݰޤ‬ᘉεϷ‫֎ڥ‬ၰᏛቚࠆ฻౜ຝǴԶ ೷ԋᄌ‫ޤ܄‬೽ว‫ݹ‬ϸᔈ37ǶLPSନΑёа‫ޔ‬ௗ೷ԋ‫ޤ‬೽ϣҜಒझ ཞ໾Ѧ38ǴΨёа೷ԋ‫֎ڥ‬ၰϷ‫ޤ‬ൻᕉ‫ޑ‬фૈ౦தǴ೭٤౦தх ࡴ‫ޤ‬Ոࢬ୏ΚБय़࡚‫ׯ܄‬ᡂϷ‫ޤ‬ಔᙃ೯೸‫܄‬ઇᚯǴԶ೷ԋߚЈӢ ‫ޤ܄‬НဍǴ຾Զ‫֎ڥ‬૰ᆃࣗԿԝΫ39Ƕ 9.

(20) ‫ޤ‬ύ֎ΕLPSǴҗLPS‫ڈ‬ᐟ‫ߞޑ‬ဦ໺ሀᐒ‫཮ڋ‬໒‫୏௴ۈ‬ว ‫ޑݹ‬ಒझᐟનϷϯᏢᖿϯӢη(chemokines)Ϸߕ๱Ӣη (adhesion moleculars)฻೷ԋύ‫܄‬ౚ(neutrophil)౽‫ډ‬ՈᆅϣҜ ಒझϷ‫ޤ‬΢ҜಒझǴ຾Զ౽Ε‫ݰޤ‬ϣ40,41Ƕว‫߻ݹ‬ϐಒझᐟન ӵtumor necrosis factor-Į (TNFĮ)ǴInterleukin-1ȕ (IL-1ȕ)Ϸ interleukin-6 (IL-6) Ǵ཮ӧ‫ޤ‬೽ว‫ݹ‬วғϐ߻εໆ‫߄ޑ‬౜42,43Ƕ ೭٤ว‫߻ݹ‬ϩ‫ݜ‬рٰ‫ޑ‬ಒझᐟનǴӵTNFĮ ϷIL-1ȕǴёа‫ڈ‬ᐟ ਡϣᙯᒵӢηNF-țB຾Εਡύ໒௴΋‫س‬ӈว‫ݹ‬໒‫߻ۈ‬΋‫س‬ӈ‫ޑ‬ ಒझᐟનౢғ44Ƕӧ΋‫س‬ӈ‫زࣴޑ‬ύว౜‫֎ڥ‬ၰ‫ޑ‬΢Ҝಒझё ӢNF-țB‫ុ࡭ޑ‬բҔ٬ள΋٤ϯᏢᖿϯӢηǴӵIL-8ǵ keratinocyte-derived chemokine (KC)ǵmacrophage inflammatory peptide-2 (MIP-2)ǵmonocyte chemotactic peptide (MCP-1)εໆ‫߄ޑ‬౜Ǵ٬ள‫ޤ‬೽ว‫ݹ‬ӢԶуख़45-47Ƕ೭ ٤Ӣη཮٬ளεໆԖᜢว‫ޑݹ‬қՈౚǴӵύ‫܄‬қՈౚǴεໆ຾ Ε‫ޤ‬೽‫ޜޑ‬໔ύǴ൞ϟқՈౚΕߟಔᙃϐΟ‫؁‬ᡯ྽ಔᙃ‫ډڙ‬໾ ্ౢғว‫ݹ‬ϸᔈਔǴՈࢬύ‫ޑ‬қՈౚ཮೏֎Ї٠ᎂ౽Կว‫ݹ‬ಔ ᙃǴаჹ‫ל‬Ѧٰ‫ނ‬፦‫ࢂ܈‬ၸ௵চǴԶ൞ϟқՈౚΕߟಔᙃϐЬ ा‫؁‬ᡯԖΟ48Ǻ ‫؁‬ᡯ΋ǵᄾ୏(rolling) ྽ಔᙃ‫ډڙ‬໾্ਔ൳ϩដϣǴύ‫܄‬ౚ཮ᆶว‫ݹ‬ೀՈᆅᏛว ғҬϕբҔǴԶӧՈᆅᏛ΢຾Չᄾ୏(rolling)ǴԜ‫؁‬ᡯҗқՈౚ ΢‫ ޑ‬L-selectin ᆶՈᆅϣҜಒझ΢‫ ޑ‬E Ϸ P-selectin ‫܌‬൞ϟǶ ‫؁‬ᡯΒǵࢲϯ(activation): ‫ܭ‬ՈᆅᏛ΢ᄾ୏‫ޑ‬ύ‫܄‬ౚ‫ֽڙ‬೽‫ੱݹ‬ϸᔈ‫܌‬ញ‫ܫ‬ϐᖿϯᐟ ન(ӵǺIL-1ȕǵMCP-1ǵMIP-2 ฻)ǵಒझᐟન(GM-CSFǴIL-8 ฻)ϷᖿϯӢη(chemoattractantsǴӵǺC5a ฻)ࢲϯԶ຾Չ‫ז‬ ೲ‫ׯ‬ᡂǴхࡴ፿ጨύ‫܄‬ౚ߄य़ L-selectinǴ٠ፓ࿯ࢲϯќ΋ௗ‫ڙ‬ Ꮤ integrin(ӵ:beta1ǵbeta2 integrin)߄౜Ǵ‫ࢲڙ‬ϯ‫ ޑ‬integrin ཮ቚமύ‫܄‬ౚᆶՈᆅϣҜಒझ่ӝ‫ޑ‬ΚໆǶ ‫؁‬ᡯΟǵᎂ౽(transmigration): 10.

(21) ՈᆅϣҜಒझ΢ߕ๱Ӣη(adhesion molecule, ӵ ICAM-1, VCAM-1)฻‫ډڙ‬ಒझᐟન(ӵǺTNF͉ǴIL-1ȕǴ IFN-ȖǴ IL-8 ฻)ࢲϯԶ߄౜ቚуǴᆶࢲϯ‫ޑ‬ύ‫܄‬ౚ΢ϐ integrin ᆙஏ่ ӝǴ٠೷ԋύ‫܄‬ౚࠠᄊ‫ׯ‬ᡂǶύ‫܄‬ౚ࿶җࠠᄊ‫ׯ‬ᡂԶऀ೸Ոᆅ ϣҜಒझቫǴவՈᆅϣᎂ౽рՈᆅѦԿว‫ݹ‬ಔᙃǶ ԖԜё‫ـ‬ǴқՈౚΕߟಔᙃ᏾ঁၸำǴନΑ‫ډڙ‬ൻᕉύᖿ ϯᐟનǵಒझᐟનϷᖿϯӢη฻ӢનቹៜѦǴᆶว‫ݹ‬ಒझ߄य़ ௗ‫ڙ‬Ꮤ(selectinǵintegrin)ϷϣҜಒझ΢‫ݠ‬ᗹϩη selectinǴ ICAM-1Ǵ฻߄౜ԖᜢǶ ќѦॶள΋ග‫ࢂޑ‬Ǵ‫ל‬ว‫ޑݹ‬ಒझᐟનȐёૈࢂᅿज़‫ڋ‬ว ‫ݹ‬ϸᔈϷว‫ݹ‬ጄൎ‫ޑ‬ಒझᐟનǴӵIL-4ǵIL-10ǵIL-12ȑ Ǵ೭٤ ‫ל‬ว‫ݹ‬ᐟનࠅό཮υᘋჹΓᡏԖ੻‫ޑ‬எЬҁ‫ޑي‬ಃ΋ጕ‫ޑ‬ٛᑇ ‫س‬಍ȐӵDefensinȑϷխࣝբҔ(ံᡏ)49Ƕ ಃΒ࿯ ‫ޤ‬೽ว‫ݹ‬ϸᔈ࣬ᜢ‫ޑ‬ಒझᐟનϟಏ(კ2.1) (΋) ߻ว‫ݹ‬ಔᙃᐟન(proinflammatory cytokine) җӚᅿᖏ‫زࣴ׉‬ᡉҢว‫ݹ‬ϸᔈวғਔǴว‫ݹ‬ಒझᐟન (inflammatory cytokines)ϷᖿϯӢη‫׎܌‬ԋፄᚇ‫ޑ‬ networkǴӧ‫ޤ‬೽ว‫ݹ‬ϸᔈวғਔ‫ת‬ᄽ๱໺ሀǵ‫ܫ‬εϷᆢ࡭ ‫ޑ‬ख़ा‫ف‬ՅǶ೭ύ໔ᗋхࡴΑ‫ڈ‬ᐟౢғ‫ל‬ว‫ݹ‬ಒझᐟન (anti-inflammatory cytokines)ᆶ߻ว‫ݹ‬ಔᙃᐟનϐ໔‫࣬ޑ‬ϕ ‫ל‬ᑽǴٰፓ࿯᏾ঁว‫ݹ‬ϸᔈ‫ޑ‬ၸำǶ߻ว‫ݹ‬ಔᙃᐟનӵIL-1ȕ ϷTNFĮǴёаӧ‫ޤ‬ว‫ݹ‬஻‫ޣ‬ϐ‫ࢱؑݰޤ‬న(bronchoalveolar lavage fluids; BALF)ύӕਔว౜Ǵߦ٬Ϸуமว‫ݹ‬ϸᔈ‫ޑ‬ว ғǴаΠ‫ॺך‬ଞჹΒ‫ޣ‬բ΋٤ϟಏǺ 1.ಃ΋қಒझᐟન(IL-1ȕ) IL-1ȕёа‫ڈ‬ᐟ೚ӭ‫ޑ‬ᖿϯӢηౢғǴӵಃΖқಒ झᐟન (IL-8)52, ൂਡౚᖿϯೈқ-1 (MCP-1)53, ϷѮᏘ 11.

(22) ಒझว‫ݹ‬ϸᔈೈқ-1Į (MIP-1Į)54Ƕ‫܌‬аIL-1ȕࢂว‫ݹ‬ϸᔈ ύว‫ݹ‬ಒझᐟનೱᙹϸᔈ (inflammatory cytokine cascade)‫ޑ‬Ӄᖿ‫ޣ‬Ƕ೚ӭࣴ‫ز‬ᡉҢࡰрӧARDS஻‫ޑޣ‬ BALF‫੆܈‬рన(edema fluid)ύёаว౜߻ว‫ݹ‬ಔᙃᐟન ‫ޑ‬Ӹӧ53-56ǶǴӕਔΨࡰрIL-1ȕࢂᆢ࡭ARDS஻‫ޤޣ‬೽ว ‫ރݹ‬ᄊ‫ޑ‬Ьा‫ޑ‬ಒझᐟન55ǶӕਔIL-1ȕΨ཮௴୏ᠼᆢ‫ޑ‬ቚ ғ(fibroproliferative)Ϸ΢Ҝ‫ޑ‬অៈၸำ(epithelial repair processes)57,58Ƕᖏ‫زࣴ׉‬ΨᡉҢIL-1ȕ೷ԋ‫ޤ‬೽࡚‫܄‬ว‫ݹ‬ ϷARDS‫ޑ‬Ԑයੰ౛ౢғ(pathogenesis)՞Ԗ࣬྽ख़ा‫ޑ‬ Տ࿼56Ƕ 2.ဍዦᚯԝӢη(Tumor necrosis fsctor-alphaǹTNFĮ) ࣬՟‫ܭ‬IL-1ȕǴTNFĮΨࢂ΋ঁࡐख़ा‫߻ޑ‬ว‫ݹ‬ಔᙃᐟ નǴѬૈ‫ڈ‬ᐟ೚ӭಒझᐟન‫ޑ‬ౢғ59-61ǶӧARDS஻‫ޑޣ‬ BALFύΨёෳ‫ډ‬TNFĮ‫ޑ‬ϲଯǴՠόૈҔԜႣෳੰ‫ޑݩ‬ᄽ ᡂ62,63Ǵՠࢂว౜ӧARDS஻‫ޣ‬ϐBALFύ‫ޑ‬TNFĮ཮Ӹӧຬ ၸ΋ຼа΢Ǵ٠Ъቹៜ‫ޑޤ‬໩ᔈ‫܄‬Ϸᝄख़‫ޑ‬ե਼Ոੱ(lung compliance and severity of hypoxemia)‫ޑ‬ౢғᐒ౗Ԗᜢ 564. Ƕё‫ـ‬TNFĮჹARDSύԖշ‫ܭ‬Ⴃෳ࣬ᜢ੯ੰౢғ‫ޑ‬ᐒ. ౗Ƕ‫܌‬аTNFĮ‫ޑ‬ෳ‫ۓ‬ჹ‫ޤ‬೽࡚‫܄‬ว‫܈ݹ‬ARDSΨࢂ࣬྽ख़ ा‫ޑ‬Ƕ. 12.

(23) Figure 2.1. The cytokine network between the different cells. 3.ᖿϯӢη ྽‫ޤ‬೽໒‫ۈ‬ౢғว‫ݹ‬ϸᔈਔǴPolymorphonuclear Leukocytes (PMN) ൩཮җՈనύӛ‫ݰޤ‬Ϸ਻ᆅ੆೸ǶӚᅿ ‫ޑ‬ϯᏢᖿϯӢη൩཮ౢғٰᇨᏤPMN຾Չ੆೸ϷӣࢬբҔ (recruitment)ǶᖿϯӢηёа୔ϩࣁΒᅿǺ΋ᅿࢂ Į-chemokines (CC) ӵMCP-1Ǵ‫ځ‬բҔёаЇᇨൂਡౚ (monocytes)ǵ T cellsǵ ༐ᡵ‫܄‬қՈౚ(basophils)Ϸ༐Ҳआ ‫܄‬қՈౚ(eosinophils)64ǹќѦ΋ᅿࢂE-chemokines (CXC) ӵಃΖқಒझᐟનIL-8 (in human), KCϷMIP-2 (in mice)Ǵ ࢂ΋ᅿ֎Їύ‫܄‬ౚ‫ޑ‬ᖿϯӢηǶIL-8ёҗΓᜪ‫ݰޤ‬ѮᏘಒझ Ϸ‫ޤ‬೽΢Ҝಒझౢғ65,66ǴӕਔΨԖࣴ‫ز‬᛾ܴӧARDS஻‫ޣ‬ ϐBALFύ֖Ԗεໆ‫ޑ‬IL-854,67Ƕ KCǵMIP-1Ϸ MIP-2 Ψࢂ ӧ‫ޤ‬೽ว‫ݹ‬ਔ࣬ᜢ‫ޑ‬ᖿϯӢη65,66Ǵӧ΋٤ࢲᡏ‫ޑ‬ჴᡍύε ໆ߄౜KCǵ MIP-2Ϸ MCP-1 ཮ගଯ‫ޤ‬೽‫ޑ‬ว‫ޑݹ‬ำࡋ 13.

(24) 68-71. ǶѬॺ཮٬‫ݰޤ‬Ϸ਻ᆅϣ‫ޑ‬PMNኧໆቚуǴӵԜߡ཮٬. ஻‫ޤޑޣ‬೯਻ᝄख़‫ޑ‬෧Ͽ,Ѐ‫ځ‬ӧARDS‫ੰޑ‬Γ54Ƕ. 4.ಃϤқಒझᐟન (Interleukine-6ǹIL-6) IL-6ࢂ΋ঁ࡚‫ޤ܄‬೽ว‫ޑݹ‬ख़ाࡰ኱ǴԖࣴ‫ࡰز‬рёа ӧ‫ޤ‬೽࡚‫܄‬ཞ໾஻‫ޑޣ‬BALFύෳளεໆ‫ޑ‬IL-6Ǵ٠Ъࡰрග ଯϷ࡭ុౢғIL-6ᆶARDSǵ௳Ոੱ(sepsis)Ϸബ໾(trauma) ஻‫ޑޣ‬ԝΫ౗Ԗᜢ72,73ǶќѦΨԖࣴ‫ࡰز‬рIL-6‫୷ޑ‬Ӣ‫ޑ‬௓ ‫ڋ‬Ӣη(promoter region)‫ޑ‬ӭࠠ‫ (܄‬polymorphisms)ёа෧ ϿIL-6‫ޑ‬ϩ‫ݜ‬ᐚࡋǴ٠Ъ෧Ͽуៈੰ‫܊‬ϣ‫ޤ‬೽࡚‫܄‬ว‫ݹ‬஻‫ޣ‬ ‫ޑ‬ԝΫ౗74Ƕ‫܌‬а෧Ͽ‫ޤ‬೽ว‫ݹ‬ਔ‫ޑ‬IL-6ౢғǴԖշ‫ޤܭ‬೽ ࡚‫܄‬ว‫࣬ݹ‬ᜢੰΓ‫ޑ‬ႣࡕǶ (Β)‫ל‬ว‫ݹ‬ಒझᐟન Ԗ٤ಒझᐟન‫ڀ‬Ԗ‫ל‬ว‫ޑݹ‬բҔǴԖ٤୏‫زࣴނ‬ᇡࣁ‫ڀځ‬ Ԗᕍਏ‫ޑ‬ወΚ75,76ǴՠҞ߻ٰ࣮ۘό‫ڀ‬ԖჴՉ‫ޑ‬ሽॶǴനԖё ૈճҔ೭‫ל‬ว‫ޑݹ‬ಒझᐟનٰ‫ݯ‬ᕍว‫ݹ‬੯ੰ‫ޑ‬БԄǴ൩Ҟ߻ٰ ᇥǴ൩ࢂว৖ёૈ‫ޑ‬ᛰ‫ٰނ‬ᇨวϣғ‫ל܄‬ว‫ݹ‬ಒझᐟન (endogenous cytokines)‫ޑ‬ញ‫ࢲ܈ܫ‬ϯ‫ל‬ว‫ޑݹ‬ಒझᐟન‫ޑ‬ௗ ‫ڙ‬ᡏ٬‫ל‬ว‫ݹ‬ϸᔈӝ౛‫ޑ‬Πफ़77ǶҞ߻ന‫ڙ‬ᡗҞ‫לޑ‬ว‫ݹ‬ಒझ ᐟનࢂIL-10ϷIL-12ǴаΠϩձբϟಏǺ 1.ಃΜқಒझᐟન(Interleukine 10ǹIL-10) IL-10 ࢂ΋ᅿ‫ڀ‬Ԗanti-inflammatory cytokineǴѬૈ‫ڋ׭‬ Ӛᅿว‫ݹ‬ೈқ‫ޑ‬ӝԋǴ೭х֖Α΋٤ಒझᐟન (TNFĮǵ GM-CSFǵIL-5ǵchemokines) Ϸ΋ᅿ཮ӧว‫ݹ‬ਔၸࡋ߄౜ ‫ޑ‬ሇન(iNOS)78ǶӧѐନIL-10 ୷Ӣ‫ޑ‬ᙯ෗ಒझ΢࣮‫ډ‬Ѩѐ IL-10 ёૈ཮೷ԋၸ௵‫܄‬੯ੰ(atopic diseases)‫ރޑ‬ᄊ78-81Ƕ IL-10 όѝ‫ڋ׭‬TNFĮϷchemokinesǴΨ཮ჹ‫୷ڋ׭‬፦ߎ 14.

(25) ឦೈқ䁙(matrix metalloproteinases)ǴӵMMP-9Ǵ‫཮ځ‬ઇᚯ ‫ޤ‬ჴ፦(lung parenchyma )ύ‫ޑ‬ቸ‫܄‬ᠼᆢ(elastin)82ǴԶIL-10 ཮ቚуಔᙃ‫୷ڋ׭‬፦ߎឦೈқ䁙(tissue inhibitors of MMPs; TIMPs)‫ޑ‬ញ‫ܫ‬Ǵ ෧Ͽว‫ݹ‬ਔჹ‫ޤ‬೽೷ԋ‫ޑ‬໾্Ƕ 2.ಃΜΒқಒझᐟન(Interleukine 12ǹIL-12)ϷಃѤқಒझᐟન (Interleukine 4ǹIL-4) IL-12 ࢂϣғ‫ޑ܄‬ፓ࿯Th1 cell ‫ޑ‬ว৖Ϸፓ௓Th1 / Th2 ‫ޑ‬ѳᑽ83Ƕ‫ځ‬ύIL-12‫ک‬IL-4ϩձࣁTh1‫ک‬Th2ಒझϩϯ‫܌‬ Ѹ໪84Ƕ‫ૈځ‬ӧ‫ܭ‬ೱௗӃϺ‫ޑ‬խࣝϷࡕϺ‫ޑ‬խࣝϸᔈ(innate and adaptive immune responses)ǴIL-12 ё‫ډڙ‬Th2ಒझ‫܌‬ ϩ‫ޑݜ‬IL-10‫ޑ‬ӣ㎸௓‫ڋ‬85ǶஒIL-12‫׫‬ԴႵёߔว‫ݹ‬ϸᔈ‫ޑ‬ว ғϷ‫ڋ׭‬ၸ௵ϸᔈ86ǶIL-12 ё٬IFN-Ȗញ‫ܫ‬ϷTಒझϩϯ87Ƕ Γπ‫ޑ‬IL-12ρ࿶ԖӧΓᡏ‫׫‬ᆶ‫ޑ‬࿶ᡍǴ‫ࢂ่݀ځ‬ౢғᝄख़‫ޑ‬ ࢥ‫܄‬ϸᔈ88ǴՠΨ࣮‫ډ‬྽஻‫ڙޣ‬ၸ௵চ‫ڈ‬ᐟਔ༐Ҳआ‫܄‬қՈౚ ܴᡉ‫ޑ‬෧Ͽ89Ƕচ‫ۈ‬TಒझाϩϯԋࣁTh2 cellሡाIL-4‫ޑ‬բ ҔǴԶЪIL-12ϷIL-4ёаߔᘐTNFĮ‫ޑ‬ϩ‫ݜ‬90Ǵ຾Զ෧጗࡚‫܄‬ ว‫ݹ‬ϸᔈ‫ុۯޑ‬Ƕࡺፓ࿯IL-4‫܈‬IL-12‫ޑ‬ϩ‫ݜ‬Ψࢂफ़եว‫ݹ‬ϸ ᔈ໾্‫ޑ‬΋ᅿБԄǶ (Ο)΋਼ϯේᆶ‫ޤ‬೽ว‫ݹ‬ϸᔈ! ΋਼ϯේӧথ٢୏‫ނ‬΢ࢂҗ΋਼ϯේ‫ޑ‬ӝԋሇન(NO! synthasesǹNOS)‫܌‬ᇙ೷‫ޑ‬ǶNOS ԖΒᅿόӕ‫ޑ‬౦ᄬሇન (isoenzymes)Ǵcalcium-dependent (cNOS)(Ξᆀࣁ endothelial constitutive NOSǹecNOS) Ϸ߄౜ӧઓ࿶ϡ‫ޑ‬ nNOS‫ک‬ಒझᐟન‫܌‬ᇨว‫ޑ‬NOS (cytokine-inducible NOSǹ iNOS)ǶcNOSӧჹ‫ޑޤ‬ᆢ࡭ᑽ‫ࡐࢂۓ‬ख़ा‫ޑ‬Ǵӵઓ࿶௓‫ޑڋ‬ Ոᆅᘉ஭(neurogenic pulmonary vasodilatation)ǵ਻ᆅᘉ஭ (bronchodilation)Ϸխࣝ‫ޑ‬ፓ࿯92-94Ƕՠࢂӧ‫ޤ‬ύ‫ޑ‬cNOS ӧ 15.

(26) ALIϷ௳ՈੱϸԶࢂ೏ፓफ़‫ޑ‬95Ǵ࣬ϸ‫ޑ‬ǴiNOS‫߄ޑ‬౜ࠅࢂё ೏ว‫ڈݹ‬ᐟӢη(ӵcytokinesǵbacteriaϷLPS)96Ƕ༐ύ‫܄‬қ ՈౚϷѮᏘಒझࢂӧALIύനЬा೏ᇨวଯໆ‫ޑ‬iNOS Ϸ‫܌‬ ౢғ‫ޑ‬NO97Ƕ ೚ӭൔ֋ᡉҢǴALI‫܈‬ARDS‫ޑ‬஻‫يޣ‬΢Ԗεໆ‫ޑ‬NO ౢ ғǴ‫ځ‬BALFύ‫ޑ‬ଯໆnitrites/nitratesჹARDS஻‫ࢂޣ‬ԖӒᓀ ‫ޑ‬98-100Ƕࡺफ़եALI‫܈‬ARDS஻‫ޣ‬BALFϐNO‫܈‬iNOSࢂԖ‫ׯ‬๓ ஻‫ޣ‬Ⴃࡕ‫ޑ‬ਏ݀98Ƕ. (Ѥ)NF-țBϷAP-1ᆶ‫ޤ‬೽ว‫ݹ‬ϸᔈ‫ޑ‬ᜢ߯ ӧ‫ޤ‬೽ว‫ݹ‬੯ੰၸำύ཮ౢғ೚ӭ‫ޑ‬ಒझᐟનϷᖿ ϯӢηǴ೭ว‫࣬ݹ‬ᜢ‫ޑ‬ᐟનࢂ‫ډڙ‬NF-țBϷAP-1‫ޑ‬ፓ௓ 101-103. Ƕ྽ว‫߻ݹ‬ಒझᐟન‫ޑ‬ௗ‫ڙ‬ᡏ‫ࢲډڙ‬ϯǴߡ཮ஒ. NF-țB-inducing kinase(NIK)ࢲϯǴNIKΞॄೢIKK-ȕ‫ࢲޑ‬ϯ (ᕗለϯ)ǴԶIțB(Inhibitor of NF-țB)‫ޑ‬ᕗለϯΞ‫ډڙ‬IKK-ȕ (inhibitors of IțB kinase-ȕ)‫܄ࢲޑ‬ፓ௓Ǵ΋ՠIțB‫ډڙ‬ᕗለ ϯǴIțB ubiquitin ligaseёஒᕗለϯ‫ޑ‬IțBௗ΢ubiquitinԶϩ ှǴӕਔNF-țB‫ډڙ‬IțB‫ޑ‬ᕗለϯࡕǴ‫ܫ‬໒NF-țBԶᙯ౽Ε ಒझਡϣǴௗ΢NF-țB‫ޑ‬promotorǴߡ໒௴ว‫ݹ‬Πෞ‫୷ޑ‬ Ӣ߄౜101ǶӧNF-țB knock-out mice‫ޑ‬ჴᡍύǴ೷ԋխࣝ ‫ڋ׭ډڙ‬ϷѨѐЬ୏խࣝ‫س‬಍(impair host defenses)Ǵ٬ ளNF-țB knock-out miceள‫ډ‬Α௳Ոੰ(septicemia)ǴҗԜ ٰ࣮௓‫ڋ‬NF-țBࢲϯ‫ޑ‬ၡ৩ჹ‫ܭ‬ว‫܄ݹ‬੯ੰ‫ޑ‬௓‫ࡐࢂڋ‬ख़ ा‫ޑ‬104ǶќѦ΋చჹว‫ݹ‬ϸᔈΨΜϩख़ा‫ޑ‬໺ሀၡ৩ࢂ Mitogen-activated protein (MAP) kinases pathway‫܌‬ᆅ௓ ‫ޑ‬p38 MAP kinase pathway102,103Ƕ ӧҗλϩη‫ޑ‬p38 MAP kinase‫ڋ׭‬ᏊǴӵSB203580, SB 239063Ϸ RWJ 67657(ς‫ޑޕ‬cytokine synthesis anti-inflammatory drugsǹCSAIDS)Ǵёаቶ‫ޑݱ‬ᓸ‫ڋ‬ว‫ݹ‬ϸᔈ105Ƕp38‫ࢲޑ‬ 16.

(27) ϯ཮٬AP-1 complex຾‫ډ‬ಒझਡϣǴࢲϯAP-1‫ޑ‬ promotorǴԶࢲϯ೚ӭว‫࣬ݹ‬ᜢ‫୷ޑ‬Ӣ105Ƕ‫܌‬а‫ڋ׭‬NF-țB ϷAP-1ᆶ‫ޤ‬೽ว‫ݹ‬ϸᔈ‫ޑ‬௓‫ڋ‬Ԗ࣬྽‫ޑ‬ᜢ߯Ƕ. 17.

(28) ಃΟക ಃ΋࿯. ‫׷‬਑ᆶБ‫ݤ‬. ‫׷‬਑. 1ǵჴᡍ୏‫ނ‬ ҁࣴ‫܌ز‬௦Ҕϐ BALB/c λႵࣁ 6 ຼεଫ໢ႵǴᖼԾ୯ࣽ཮ჴ ᡍ୏‫ނ‬ύЈǶ 2ǵགࢉচ ϣࢥન LPS (Escherichia coli, 0055:B5)ᖼԾ Sigma Chemicals (St. Louis, MO)Ƕ 3ǵ၂Ꮚᛰࠔ ၂ᏊӜᆀ. ᇙ೷Ϧљ. Ҟᒵጓဦ. Acetic acid. MERCK. 100063. Acrylamide. Bio-Rad. 161-0101. Agarose. Seakem. 1-800-341-1574. Albumin. Sigma. A-4919. Ammonium persulfate. Bio-Rad. 161-0700. Bis-acrylamide. Bio-Rad. 161-0201. Chloralhydrat. MERCK. K24127725. Citric acid. Sigma. C-0759. Coomassie Brilliant blue. Bio-Rad. 137385A. Eosin. Sigma. E4382. Fetal Bovine serum. Hyclone. SH30071.03. Glycerol. MERCK. K22855994. Glycine. MERCK. K22855994. Griess reagent. Sigma. G441-10G. HCl. Sigma. H-7020. Hematoxylin. Sigma. H3136. Riedel-de Haen. 18312. H2O2 Ketamine. MERIAL 18. W099062B.

(29) LightShiftTM. PIERCE. 89880. TEDIA. MS-1922. PIERCE. 23235. Nuclear Extraction Kit. Panomics. AY2002. One-step RT-PCR kit. GeneMark. RP01. Paraformaldehyde. Sigma. P-6148. PBS. Sigma. D5652. Periodic acid. Sigma. P7875-25G. 2-Propanol. Sigma. I9516. Schiff’s reagent. Sigma. S5133-500ML. SDS. Sigma. L5750. Sircol. S1111. Sodium nitritre. Sigma. S2252-500G. Sodium metabisulfite. Sigma. S-1516. Sodium iodate. Sigma. S-4007. TEMED. Bio-Rad. 7-759B. Tris. USB. 75825. Triton X-100. Sigma. 16H1421. Trizol. Invitrogen. 15596. Trypan blue. Sigma. T8154. Tween. 20. Sigma. P2287. Xylazine. Chanelle. LA1136. Xylene. TEDIA. XR-2250. Chemiluminescent EMSA Kit Methanol Micro BCA protein Assay kit. Sircol soluble collagen assay. 4ǵ‫ל‬ᡏ ‫ל‬ᡏӜᆀ. ᇙ೷Ϧљ. Mouse IL-1ȕ ELISA kit. eBioscience. Mouse TNFĮ ELISA kit. BD 19.

(30) Mouse IL-6 ELISA kit. eBioscience. Mouse IL-4 ELISA kit. eBioscience. Mouse IL-10 ELISA kit. eBioscience. Mouse IL-12 ELISA kit. eBioscience. Mouse KC ELISA kit. R&D. IțB Ϸ IțB phosphorylation Ab. Cell signal. p38 Ϸ p38 phosphorylation Ab. Cell signal. ERK Ϸ ERK phosphorylation Ab. Cell signal. JNK Ϸ JNK phosphorylation Ab. Cell signal. ICAM-1 FITC Ab. eBioscience. VCAM-1 PE Ab. eBioscience. Streptavidin-HRP conjugate. BD. TMB substrate reagent set. BD. 5ǵ઻‫׷‬ ઻‫׷‬Ӝᆀ. ᇙ೷Ϧљ. 1ml ༟ጤ‫ݙ‬৔ଞ฿. Top plastic syringe. Ҟᒵጓဦ 2K05G. 23G Needle. TERUMO. 997801D. 15 ml ᚆЈᆅ. NUNC. 8-0000-560302. 50 ml ᚆЈᆅ. CORNING. 25330-50. 0.22͔m filter. Millipore. SCGPS05RE. Coating slide. Microslide. 5126. Uncoating slide. Microslide. 5156. ࡬᜔䵱. Kimble-clark. 2F083B86. 3mm filter paper. Whatman. 3030917. PVDF membrane. Immobilon-p. IPVH00010. Nylon membrane. Hybond-P. S/1282/7307/98/03. 6ǵሺᏔ ሺᏔӜᆀ. ቷจ/Ϧљ 20.

(31) ೈқ፦ႝ‫ݚ‬ኲ. Miniprotein III Cell 165-2940, Bio-Rad. ႝྍ‫ٮ‬ᔈᏔ. Bio-Rad Power Pac 300 Power Supply. ႝ‫ݚ‬ኲ. Mini-protein II Bio-Rad, U.S.A.. ࢬԄಒझሺ. Becton-Dickinson Immunocytometry system, San Jose, CA, USA. ёፓԄ᎜ᕏᏔ. Vortex mixer, model VM-1000. ଯᓸྐ๵ᗙ. Tomin, Taiwan. ଯೲհএᚆЈᐒ. Himac CR 21F, HITACHI, JAPAN. ਫ΢ࠠଯೲհএᚆЈᐒ. Beckman GS-15R, USA. ᚆЈᐒ. KUBOTA 5010, JAPAN. ᡉ༾᜔. NIKON Type 104, JAPAN. ༾ໆϺѳ. AȿD company. -20ʚհএӇጃ. SANYO, JAPAN. -30ʚհএӇጃ. Medical Freezer, SANYO, JAPAN. -80ʚհএӇጃ. REVCO, GS Laboratory Equipment ULT2586-9-030 Asheville, N.C., U.S.A.. ሇનխࣝϩ‫݋‬ղ᠐ሺ ϸᙯᒵ䁙᜘ϸᔈᏔ ค๵ᏹբѠ. DYNATECH Laboratories, Great Britain Mastercycler personal, Germany . Wintvcoon, Chin Tycoon Enlerprises Co.LTD. ѐᚆηНᇙ೷ᐒ. Modulab. ՈౚीኧᏔ. MARIENFELD, Japan. հএଳᔿᐒ. Labconco Corporation, Kansas city, Missouri, U.S.A.. Cytospin. KUBOTA , JAPAN. ֡፦Ꮤ. KINEMATICA, switzerland. НѳԄႝ‫ݚ‬ኲ. HORIZON 11.14, Life Techologies, U.S.A.. Milli-Q. Wellbond science CO., LTD. 7ǵሌᙋණ‫ޑ‬ᇙഢ. 21.

(32) 7-1ǵሌᙋණಔԋᆶК‫ٯ‬ ύЎӜᆀ ߎሌ޸. Ꮲ. Ӝ. К. ౗. Lonicerae japonica thumb.. 10. ೱᙋ. Forsythiae suspense thumb.. 10. ᖓ಻. Menthae Haplocalycis Labiatae. 4. ૘޺ᕞ. Schizonepetae tenuifolia Benth.. 4. ఩‫ل‬೟. Glycine max L.. 5. (Semen Preparatum Sojae), Ғ૛. Glycyrrhizae glabra L.. 5. ਧఒ. Platycodi grandiflorum Jacq.. 6. ఩Ԯယ. Lophatheri Gracilis Gramineae. 5. Ф⣢η. Arctii lappa L.. 6. ᝳਥ. Phragmitis communis L.. 10. 7-2ǵᛰ‫ྗނ‬ഢ БᏊᇙഢ‫࢒ૼہ‬ၲᇙᛰቷᇙഢృᆯǴᙁൂࢬำӵΠ: ߎሌ޸(Flos Lonicera)Ϸਧఒ(Radix Platycodi) ણ࿗ԋࣁ ಒણǴၸ 120 Ҟಒᑔǹᖓ಻ (Herva Mentha)Ϸ૘޺(Herba Schizonepeta)Ҕචว‫ݨ‬ᆶНྋనϩձ੆‫ݰ‬ǴӚԏ໣నᡏቫഢ ҔǴӆஒӚձූ෦ԏ໣ᆶ‫ځ‬ѬϤᅿύᛰషӝǴ‫ܫ‬ΕНύዖฆΒ ԛǴ‫؂‬ԛΒλਔǴӚԛԏ໣‫ځ‬నᡏቫǴၸᛇѐନූ෦Ƕၸᛇన ӆᆶᖓ಻Ϸ૘޺੆‫ݰ‬నషӝǴӆᆶߎሌ޸Ϸਧఒણ҂షӝ֡ ϬǴଳᔿԋࣁણ҂ǴӆၸಒᑔǶԜਔሌᙋණ‫ޑ‬ણ҃ԋࠔଌΕჴ ᡍ࠻Ǵ‫ॺך‬ஒ‫ځ‬ણ҂ྋ‫ܭ‬НύǴӧ‫ ڗ‬1 ml ગख़ࡕհএଳᔿǴी ᆉଳख़Ƕϐࡕีញԋ 100 mg/mlǴᓯપ‫ܭ‬-30ʚӇጃύഢҔǴҔ ‫ܭ‬ԴႵჴᡍҔǶ ќѦҔ‫ܭ‬ಒझჴᡍύਔǴ‫ॺך‬ஒρྋ‫ܭ‬Нύ‫ޑ‬ሌᙋණǴᚆ Ј 6000 ᙯǴ30 ϩដǴ‫ڗ‬рНቫǴҔ 0.22 ȝ Filtor բค๵ၸᛇǴ ค๵ၸᛇన, ӧ‫ ڗ‬1 ml ગख़ࡕհএଳᔿǴीᆉଳख़Ƕϐࡕีញ 22.

(33) ԋ 100 mg/mlǴᓯપ‫ܭ‬-30ʚӇጃύഢҔǴҔ‫ܭ‬ಒझჴᡍҔǶ ಃΒ࿯. Б‫ݤ‬. 1ǵLPS ᇨว‫ݹޤ‬Ⴕϐ୏‫ނ‬ኳԄࡌҥ 1-1ǵ਻ᆅϣௗᅿȐintratracheal inoculationȑ Ȑ1ȑ ǵ၂Ꮚᇙഢ ഞᎈᏊ ketamineǺxylazineɨ2Ǻ1ǴҔค๵ 1X PBS ีញ 10 ७Ǵ‫؂‬ଫλႵ๏ϒऊ 0.24-0.3 mlǴ‫ݙ‬৔‫ډ‬λ Ⴕဎ๚Ƕ Ȑ2ȑ ǵ‫؁‬ᡯ λႵ೏ഞᎈࡕǴஒϐࢎӧࡼѺ਻ᆅϣ‫ݙ‬৔‫ޑ‬πբ Ѡ΢ǴѰЋε‫کࡰܠ‬१ࡰᇸᓸλႵࡕတ೽ǴஒλႵΠ ߐᏁԌ‫៓ډ‬ጕ΢ǴѓЋҔᢍη֨рλႵ‫ޑ‬ՁᓐǴа༾ ໆ‫ݙ‬৔Ꮤӧോᝁೀᅀ຾ 50͔l LPSȐ1 mg/mlȑ Ƕࡑλ Ⴕ֎Ε LPS ྋనǴวр༙ሷᖂࡕ᚞໒ᢍηǴ‫ܫ‬௞Ձ ᓐǴλႵߥ࡭‫ޔ‬ҥ‫ރ‬ᄊ 1 ϩដࡕǴ࿼‫ܭ‬ᘚᐩ߯Πߥ ྕǴ‫ࡠډޔ‬ൺమᒬǶ 1-2ǵՈమϩᚆ ഞᎈλႵǴഞᎈᏊ 0.3 ml/ଫǴࡑλႵॹΠǴҔଞᓐ ଝӧߥ᜽ᓪ݈΢Ǵа 75% ଚᆒቔ‫ૅځ‬ဎ೽ǴவѰ๛Π୊ ໒ҜЛϷૅεԼǴ‫ډפ‬๛Π୏ે‫ܫ‬ՈǶа 1 ml ଞ฿ӣԏ Ոనऊ 1 mlǴ ‫ݙ‬Ε eppendorf Ǵࡑ࠻ྕᓉ࿼ 1 λਔࡕǴ а 4ʚǴ3000 rpmǴ20 ϩដǴԏ໣΢మన֡ϩ 3-4 eppendorf Ǵ‫؂‬ᆅ 100͔lǴՈమߏයߥӸ‫ܭ‬-80ʚǶ 2ǵЍ਻ᆅ‫ࢱؑݰޤ‬నϐಒझᏢᔠෳ Ȑ1ȑ ǵ၂Ꮚᇙഢ 0.5ʝFBS/1xPBS 1ØPBS. 50 ml 23.

(34) Fetal bovine serum. 250͔l. Ȑ2ȑ ǵ‫؁‬ᡯ λႵԏ໣ՈనኬҁࡕǴฅࡕஒλႵဎ೽ර΢ ‫ۓڰ‬ӧှও݈΢ǶҔ 75%ଚᆒ੃ࢥӄ‫ي‬ҜЛǴஒΠ ᚓԿૅ๚‫ޑ‬Ҝጥ୊໒Ǵ‫ע‬ൄనဏϷᓍ೽ԼԺቫ܎໒ ኪ៛਻ᆅǴ୊໒਻ᆅ΢‫ޑ‬ᖓጢǴϐࡕӧ਻ᆅ΢࿹΢ ᕳጕǴӧ਻ᆅ᎞߈ോോᓐೀϪ໒΋໒αǴකΕ 0.965 mm ‫ ޑ‬polyethylene Tubing ҃ᆄௗ΢ 25G ‫ޑ‬ଞᓐ ঺ᆅǴుΕഄ 0.5 cmǴ࿹ᆙᕳጕஒ঺ᆅ‫ۓڰ‬ӧ਻ᆅ ΢ǶஒးԖ 1 ml 0.5ʝFBS/1xPBS ‫ޑ‬ଞ฿ක΢ଞ ᓐǴᄌᄌѺ຾‫ޤ‬೽Կ᏾ঁ‫ޤ‬೽ֹӄᑩ๞Ǵӆӣ‫ܜ‬Ǵ ӵԜϸᙟΟԛࡕǴஒፂࢱనߥӸ‫ܭ‬ӇύǶӆඤќѦ ΋Ѝ 1 ml 0.5ʝFBS/1xPBS ‫ޑ‬ଞ฿Ǵख़ᙟ߻ॊ‫؁ޑ‬ ᡯǶஒԏ໣‫ࢱؑޑ‬న(ऊ 1.8 ml)Ǵа 1200 rpmǴ4 ʚǴ10 ϩដᚆЈǴԏ໣΢మన‫ܭ‬Ǵ-20ʚߥӸǶಒ झӆуΕ 1 ml 0.5ʝFBS/1xPBS ‫ޑ‬నӣྋǶ 2-1ǵ਻ᆅ‫ࢱؑݰޤ‬నύಒझᕴኧ Ȑ1ȑ ǵ၂Ꮚᇙഢ Trypan blue ีញ 4 ७. Ӛ 40͔l. 1xPBS. Ӛ 20͔l. Ȑ2ȑ ǵ‫؁‬ᡯ ‫ ڗ‬20͔l ‫ࢱؑݰޤ‬నԿ eppendorfǴуΕ 20͔l 1xPBS ‫ีک‬ញ 4 ७‫ ޑ‬Trypan blueǴ֡Ϭష ӝࡕа hemocytometer ीᆉಒझኧҞǶ 2-2ǵ਻ᆅ‫ࢱؑݰޤ‬నύқՈౚϩᜪीᆉ Ȑ1ȑ ǵ၂Ꮚᇙഢ LiuA Ϸ LiuB 24.

(35) Ȑ2ȑ ǵ‫؁‬ᡯ ‫ ڗ‬100͔l Ѝ਻ᆅ‫ࢱؑݰޤ‬నǴа cytospin ஒ ಒझѺԿ࣒Т଺ԋಒझ‫ܓ‬ТǴ॥ଳࡕǴа Liu ࢉ నࢉՅǶᅀ 3 ᅀ‫ ޑ‬LiuB ࢉనֹӄᙟᇂ‫ܓ‬ТǴբҔ 90 secǴό࿶НࢱǴӆᅀ 6 ᅀ‫ ޑ‬LiuA ࢉన 40 ࣾǴ ᙖ֌॥షӝ‫ٿ‬ᅿࢉన٠և౜ߎឦӀᐛǴբҔ 1 ϩ ដǴаԾٰНؑࢱࡕ॥ଳǴճҔ‫᜔ݨ‬Ȑ400xȑ᜔ ჸ 100 ঁқՈౚಒझᅿᜪǴीᆉӚ՞‫ޑ‬ԭϩКǴ ४΢Ѝ਻ᆅ‫ࢱؑݰޤ‬నύಒझᕴኧҞǴளрӚқ Ոౚ‫๊ޑ‬ჹኧҞǶ. 3ǵಔᙃࢉՅϐಒझᢀჸȐHematoxylin-eosin stainȑ Ȑ1ȑ ǵ၂Ꮚᇙഢ 4% paraformaldehyde milliQ Н. 500 ml. paraformaldehyde. 4g. NaH2PO4Ǹ2H2O. 6g. NaOH. 2g. Ȑ2ȑ ǵ‫؁‬ᡯ ಔᙃϪТ‫ޑ‬ᇙբǺஒλႵ‫ޤ‬೽Ϸ਻ᆅ‫ڗ‬Π ࡕǴ‫ ܭ‬4% paraformaldehyde ‫ۓڰ‬నߥӸǶ࿶ၸᅌ ຾ԄଚᆒಥНϷҡᛪхশࡕǴаϪТᐒϪԋ 0.5 mm ࠆࡋ‫ޑ‬ᖓТǴஒϐ‫࣒ܭۓڰ‬Т΢Ǵа᝵Еለ‫ک‬Ҳआ ٰࢉՅǶ 4ǵOOjusjuf ֖ໆෳ‫ۓ‬ Ȑ1ȑ ǵ၂Ꮚᇙഢ Griess reagent Sodium nitrite nitrate reductase 25.

(36) NADPH Ȑ2ȑ ǵ‫؁‬ᡯ Ӄу sample ᆶ nitrate reductase (610 mU/ml) Ϸ NADPH (170 mmol/L) షӝǴ࿼‫ྕ࠻ܭ‬ΟλਔǴ а sodium nitrite ΋‫س‬ӈีញࡕ྽ standardǴӆ‫ ڗ‬100 ͔l ‫ࢱؑݰޤ‬నǴуΕ฻ໆ Griess reagent ‫ ܭ‬96 Ͼ ѳۭ༾ໆዬǴᓉ࿼ 15 ϩដǴа ELISA reader ‫ݢܭ‬ ߏ 540 nm ୀෳ Nitrite ᐚࡋǶ 5ǵFMJTB ϩ‫݋‬ȐEnzyme-linked immunosorbent assayȑ Ȑ1ȑ ǵ၂Ꮚᇙഢ aǵwashing solutionǺ0.05% PBS-T ‫ޑ‬ଛᇙБ‫ݤ‬ 1XPBSȐPH=7.4ȑ. 500 ml. Tween-20. 250͔l. bǵcoating buffer ‫ޑ‬ଛᇙБ‫ݤ‬Ǻ Na2CO3. 1.59 g. NaHCO3. 2.93 g. D.D.H2OȐ੃ࢥၸȑ. 1000 ml. cǵblocking bufferǺ 3% BSAȐ3 g BSA/100 ml D.D.Wȑ dǵstopping buffer 1 N H3PO4 Ȑ2ȑ ǵ‫؁‬ᡯ aǵ AgȐAbȑа coating buffer ଛԋ 2͔g/mlǴа 50͔ l/well уΕ 96-well assy plateǴ4ʚΠᓉ࿼΋ఁǶ bǵ ॹр΢మనȐёӧર΢ҔΚᄟ΋ᄟȑ Ƕ cǵуΕ 3% BSA blockingǴ100͔l/well ࠻ྕ 1.5-2 λ ਔа΢ dǵ ॹ௞Ǵ‫ܡ‬ଳǴа 0.05% PBS-T wash 3 ԛǴ‫ܡ‬ଳǶ eǵ уΕࡑୁ AbȐAgȑ Ǵ100͔l/wellǴ37ʚȐRTȑ Ǵ2 λ 26.

(37) ਔ fǵ ॹ௞Ǵ‫ܡ‬ଳǴа 0.05% PBS-T wash 3 ԛǴ‫ܡ‬ଳ gǵ уΕΒ‫ ל‬Ȑ100͔l/wellȑǴ37ʚǴ45 ϩដ hǵ ॹ௞Ǵ‫ܡ‬ଳǴа 0.05% PBS-T wash 4 ԛǴ‫ܡ‬ଳ iǵ у SAV-HRP Ȑstreptoavidin- Horse radish peroxidseȑ100͔lȐ2͔g/wellȑ Ǵ࠻ྕΠᓉ࿼ 30 ϩ ដǶSAV-HRPȐ1% BSA dilute 1000xȑ jǵ ॹ௞‫ܡ‬ଳǴа 0.05% PBS-T wash 5 ԛǴ‫ܡ‬ଳ kǵуΕ substrate TMB 100͔l/wellǴևՅኧϩដǶ lǵ уΕ 50͔l/well 1 N H3PO4 stopping mǵ а ELISA reader 450 nm Π᠐ OD ॶǶ 6ǵਡೈқϩ‫݋‬ 6-1ǵਡೈқ๧‫ڗ‬ Ȑ1ȑ ǵ၂Ꮚᇙഢ Buffer A MixǺ 1X Buffer A. 1.65 ml/ᆅ. 100mM DTT. 16.5 ͔ l/ᆅ. Protease Inhibitor Cocktail. 16.5 ͔ l/ᆅ. 10% IGEPAL. 66 ͔ l/ ᆅ. Buffer B MixǺ 1X Buffer B. 147͔l/ᆅ. Protease Inhibitor Cocktail. 1.5͔l/ᆅ. 100 mM DTT. 1.5͔l/ᆅ. Ȑ2ȑ ǵ‫؁‬ᡯ λႵѰ‫ޤ‬ಔᙃу Ε 1.5 ml ‫ ޑ‬Buffer A Mix ࡕ֡፦ ϯǴ࿼‫ܭ‬Ӈ΢15 ϩដǴϐࡕ‫ ܭ‬850 xg Ǵ 4к ᚆЈ 10 ϩ ដǴѐ௞΢మనǴ ӆ࿼‫ܭ‬Ӈ΢Ǵ ٠уΕ 250͔l ‫ޑ‬Buffer A Mix ࡕ֡፦ϯǴ࿼‫ܭ‬Ӈ΢ 15 ϩដ ࡕ 15000 G Ǵ 4к ᚆ 27.

(38) 3 ϩដǶᓯӸ΢మనǴќу 150 ͔ l ‫ ޑ‬Buffr B Mix ؑණ pellet Ǵ vortexing 10 ࣾǴฅࡕ٬ eppendorf tube ѳ፴ ӧӇ΢ǴճҔrocking platform འਗ 200 rpm Ǵ 2 λਔǶ ‫ܭ‬4к ᚆЈ 15000 G Ǵ 5 ϩដǴԏ໣΢మనջࢂ‫܌ॺך‬ ा‫ޑ‬nuclear extract ǴӸ‫ ܭܫ‬-80ʚǶ 6-2ǵೈқ፦ᐚࡋ‫ۓޑ‬ໆ Ȑ1ȑ ǵ၂Ꮚᇙഢ Working reagent ȐMicro BCA protein Assay kitǹ PIERCEȑ Ȑ2ȑ ǵ‫؁‬ᡯ ‫ڗ‬рBSA 70͔lаD.D.H2Oೱុีញԋ 200ǵ40ǵ20ǵ 10ǵ5ǵ2.5ǵ1ǵ0.5ǵ0͔g/ml BSA኱ྗనǴ‫ڗ‬όӕᐚࡋ ‫ޑ‬኱ྗన‫܈‬ኬࠔ 150͔lǴуΕ 96 well plateύǴᆶ 150 ͔l Working reagentషӝǴའ 30 ࣾǴ37ʚ 2 λਔǴplate reader OD 570 nmᆉрኬࠔ‫ޑ‬ჴሞᐚࡋǶ 6-3ǵSDS-PAGE ϐೈқ፦ႝ‫ݚ‬ (1)ǵ၂Ꮚᇙഢ ှ‫݋‬ጤᡏ(separation gel) 30% acrylamideȿ0.8% bis-acrylamide. 3.3 ml. 1.5 M Tris-HCl pH8.8. 2.5 ml. 10% SDS. 1.0 ml. D.D H2O. 4.0 ml. 10% ammonium persulfate. 0.1 ml. TEMED. 0.005 ml. ᠄ቫጤᡏ(stacking gel) 30% acrylamide ȿ 0.8% bis-acrylamide 1.0 M Tris-HCl pH 6.8 10% SDS. 0.67 ml 2.5 ml 0.04 ml. 28.

(39) D.D.H20. 2.7 ml. 10% ammonium persulfate TEMED. 0.04 ml 0.004 ml. ႝ‫ݚ‬጗ፂన(running buffer) 0.025 M Tris-HCl pH 8.3 0.192 M glycine 0.1% SDS ଓᙫࢉᏊ(4X Sample buffer) 0.25 M Tris-HCl pH 6.8 8% SDS 0.02% BPB 30% Glycerol ࢉՅన(Stain buffer) 0.25% Coomassie blue R-250 10% acetic acid 45% Ethanol ಥՅన(Destain buffer) 7% acetic acid 25% Ethanol Ȑ2ȑ ǵ‫؁‬ᡯ ஒှ‫݋‬ጤᡏуΕаಔӝӳ‫ޑ‬ႝ҉း࿼Ǵӆ጗጗у ΕϿ೚ methanol ‫ځܭ‬ᆶ‫਻ޜ‬ௗ᝻ϐࣚय़ǶࡑᏉ‫ࡕڰ‬Ǵ֎ ௞ methanolǴӆуΕ᠄ቫጤᡏ٠කӳᏁ‫݈ރ‬ȐcombȑǶ ஒೈқ፦ྋనᆶ฻К‫ޑٯ‬ଓᙫྋనషӝࡕǴ‫ ܭ‬100ʚΠу ዗ 5 ϩដǴӆа‫ ঁ؂‬well loading 10͔l ‫ࡑޑ‬ෳೈқ፦ྋ నǶ೯ႝ 120 VǴ1.5 λਔႝ‫ݚ‬ϩ‫݋‬Ƕႝ‫ֹݚ‬ԋࡕǴஒှ ‫݋‬ጤᡏ‫ڗ‬раࢉՅనࢉՅǴ‫ ܭ‬50ʚΠ᎜ᕏ 30 ϩដǴӆ аಥՅనѐନङඳǶ. 29.

(40) 6-4ǵՋБᏀᗺ‫ݤ‬ Ȑ1ȑ ǵ၂Ꮚᇙഢ ᙯᅄ጗ፂనȐtransfer bufferȑ 0.192 M glycine 0.0013 M SDS 10-20% methanol 0.025 M Tris (pH 8.3) ࢉՅన 0.1% amido black 45% methanol 10% acetic acid ಥՅన 50% methanol 10% acetic acid PBS 0.8% NaCl 0.02% KCl 0.24% KH2PO4 (pH 7.2-7.45) PBST PBS Ѧу 0.05% Tween. 20 Ȑ2ȑ ǵ‫؁‬ᡯ ஒֹԋ‫ ޑ‬SDS-PAGE ጤᡏᆶ methanol ߻ೀ౛‫ޑ‬ PVDF membrane ੆‫ܭ‬ᙯᅄ጗ፂనύ 10 ϩដǶஒጤᡏ PVDF membraneǵᘠર҅ዴӦ᠄‫ܭܫ‬ႝ‫ݚ‬ᙯᅄኲύǴ٠а 100 mA ೯ႝᙯӑ 1 λਔǶPVDF membrane ੆‫ ܭ‬5% non-fat milk/PBSǴ4ʚΠᓉ࿼ၸ‫ڹ‬Ƕ‫ڗ‬р PVDF membraneǴа PBST మࢱ 3 ԛ 5 λਔࡕǴуΕྋ‫ ܭ‬PBST ࿶፾྽ีញ‫לޑ‬Ոమ(΋ ԛ‫ל‬ᡏ)Ǵ37ʚΠϸᔈ 2 λਔǶPBST మࢱ 3 ԛ 5 ϩដǶуΕ ೱ่Βԛ‫ל‬ᡏǴ37ʚΠϸᔈ 2 λਔǴPBST మࢱ 2 ԛࡕǴPBS. 30.

(41) మࢱ 2 ԛǶനࡕа ECL kit ևՅǶ 6-5ǵElectrophoretic Mobility Shift Assay (EMSA) Ȑ1ȑ ǵ၂Ꮚᇙഢ Oligodeoxynucleotide NF-KBp (specific probe) NF-kBc (specific competitor) 6% Nondenaturing polyacrylamide gel 10X TBE. 1 ml. 30% Acrylamide. 4 ml. 80% glycerol Water. 625 ml 14.375 ml. 10% APS. 300 ȝl. TEMED. 20ȝl. Binding Reagents (Light Shift Chemiluminescent EMSA kit) Transfer binding reagent 0.5X TBE buffer Detect Biotin-labeled DNA Reagents LightShift Blocking Buffer LightShift Wash Buffer LightShift Stailized Streptavidin-Horseradish Peroxidase conjugate LightShift Substrate Equilibration Buffer Ȑ2ȑ ǵ‫؁‬ᡯ Ӄଛ 6% Nondenaturing polyacrylamide gelǴӆஒଛ࿼ ӳ‫؂ޑ‬ᆅ 20͔1 binding reagentуΕ 5͔l‫ ޑ‬5X Loading BufferǴ loading ࡕ 120 V າ ऊ 90 ϩ ដ Ǵ ஒ Gel ‫ ڗ‬р ࡕ ᙯ ᅄ ‫ ډ‬nylon membraneǴmembraneа 120 mJ/cm2ኪ៛‫ ܭ‬254 nm UVᐩΠǴ ྣ৔ 2 ϩដǶஒጢ੆‫ ܭݰ‬20 ml LightShift Blocking BufferǴ጗ᄌ Ӧ འ ਗ 15 ϩ ដ Ƕ ӆ ஒ LightShift Blocking Buffer ॹ ௞ Ǵ у Ε 31.

(42) conjugate/blocking buffer solutionǴ጗ᄌӦའਗ 15 ϩដǶஒጢ‫ڗ‬ рඤԿଳృ‫ޑ‬৒ᏔϣǴа 20 ml 1X wash solutionའਗమࢱ 4 ԛǴ ‫؂‬ԛమࢱ 5 ϩដǶϐࡕӆས‫܊‬ύуΕ 30 ml LightShift Substrate Equilibration BufferǴ጗ᄌӦའਗ 5 ϩដǶ‫ډ‬௞ࡕஒLightShift Substrate Working Solution‫ޔ‬ௗуӧጢ΢Ǵϸᔈ 5 ϩដǶѐ௞ Working Solution Ǵ ஒ ጢ ‫ ܫ‬Ε ༟ ጤ ೓ ϣ Ǵ ᓸ р ӭ Ꭹ ‫ ޑ‬Working Solution຾ՉᓸТǶ 7ǵRNA ϩ‫݋‬ 7-1 RNA ๧‫ڗ‬ Ȑ1ȑ ǵ၂Ꮚᇙഢ Trizol Chloroform Isopropyl alcohol DEPC water Ȑ2ȑ ǵ‫؁‬ᡯ ஒλႵѓ‫֡ޤ‬፦Ǵ ၸ 26G ଞǴᓉ࿼ 5 ϩដǴуΕ 0.2 ml ChloroformǴᓉ࿼ 15 ϩដǴ12000 G 15 ϩដ 4ʚǴ‫ڗ‬΢మనǴ уΕ 0.5 ml isopropyl alcohol ᓉ࿼ 10 ϩដ 4ʚǴ20000 G. 20 ϩ ដ 4ʚᚆЈǴѐନ΢మనǴԏ໣؈ᐘ‫ܡނ‬ණǴа 0.7 ml 100%ଚᆒ ‫ ک‬0.3 ml DEPC water మࢱǴ20000 G. 20 ϩដ 4ʚᚆЈǴѐନ ΢మనǴ‫ ܭ‬Flow ύබ 45 ϩដǴа 20͔l DEPC water ӣྋǴෳ RNA ᐚࡋǴፓ᏾നಖᐚࡋ 1͔g/mlǶ 7-2ǵOne-Step RT-PCR Ȑ1ȑ ǵ၂Ꮚᇙഢ One-Step RT-PCR KitȐGeneMarkȑ Nuclease free H20. 15.6͔l. RNA temple. 0.6͔l. Upstream primerȐ50-100ng/͔lȑ. 0.6͔l. 32.

(43) Downstream primer (50-100ng/͔l). 0.6͔l. (ӵ߄ 3.1) 5X Reaction Mix. 6͔l. 5X Enhancer. 6͔l. Enzyme Mix. 0.6͔l Total. 30͔l. Ȑ2ȑ ǵ‫؁‬ᡯ First strand. cDNA synthesis Ǻ. 1 cycle Reverse transcription. 50ʚ, 30 ϩដ. 1 cycle M-MLV RTase inactivation. 94ʚ, 2 ϩដ. RNA/cDNA/Primer denaturation Second strand cDNA synthesis and PCR reactionǺ 35 cycle 94ʚ, 30s-1 ϩដ. Denaturation. 1 cycle. Annealing. 55-65ʚ, 30s-1 ϩដ. Extension. 72ʚ, 30s-3 ϩដ 72ʚ, 7 ϩដ. Extension. 7-3ǵAgarose gel electrophoresis Ȑ1ȑ ǵ၂Ꮚᇙഢ Agarose gel. 1.5 g. 0.5Ø TBE. 100 ml. 10 mg/ml Ethidium bromide 6Ø gel loading buffer Ȑ2ȑ ǵ‫؁‬ᡯ ‫ ڗ‬1.5 g ‫ ޑ‬agaroseǴॹΕᒷ‫׎‬౟Ǵߥᗲጢ࠾αǴ‫ڈ‬ኧঁ ࢰǴ‫ܫ‬຾༾‫ݢ‬᝗Ǵу዗ 1.5 ϩដǴॹΕႝ‫ݚ‬ኲύᏉ‫ڰ‬Ǵ጗጗уΕ 0.5Ø TBE ጗ፂྋనǶ‫ ڗ‬10͔l DNA Sample уΕ 2͔l ‫ ޑ‬6Ø gel 33.

(44) loading bufferǴуΕႝ‫ݚ‬ጤТпኲύǴ100 V 1 λਔǴ‫ڗ‬рႝ‫ݚ‬ ጤТ‫ܫ‬Ε Ethidium bromide 30 ϩដǴӆ‫ܫ‬ΕНύ຾ՉಥՅ 10 ϩ ដǴҔ UV Ӏᐟว٠‫ྣܡ‬Ƕ Table 3.1 Oligonucleotide primerdesignation for RT-PCR analysis ȕ-actin. 5’-TGGAATCCTGTGGCATCCATGAAAC-3’ 5’-TAAAACGCAGCTCAGTAACAGTCCG-3’. TNFĮ. 5’-AGCCCACGTCGTAGCAAACCACCAA-3’ 5’-ACACCCATTCCCTTCACAGAGCAAT-3’. IL-1ȕ. 5’-GCGGACTACTATGCTAAAGATG-3’ 5’-GTTGTGTTGGTTGTAGAGGGCA-3’. IL-10. 5’-GGACTTTAAGGGTTACTTGGGTTGCC-3’ 5’-CATTTTGATCATCATGTATGCTTCT-3’. iNOS. 5’-AATGGCAACATCAGGTCGGCCATCACT-3’ 5’-GCTGT-GTGTCACAGAAGTCTCGAACTC-3’. COX-2. 5’-GGAGAGACTATC-AAGATAGT-3’ 5’-ATGGTCAGTAGACTTTTACA-3’. TGF-ȕ1. 5’-GCGGACTACTATGCTAAAGATGT-3’ 5’-GTTGTGTTGGTTGTAGAGGGGCA-3’. MCP-1. 5’-ACCAGCCAACTCTCACTGAAGC-3’ 5’-CAGAATTGCTTGAGGTGGTTGTG-3’. MIP-2. 5’-GAACAAAGGCAAGGCTAACTGA-3’ 5’-AACATAACAACATCTGGGCAAT-3’. 8ǵಒझ୻ᎦϷෳໆಒझғߏϷᛰ‫܄ࢥނ‬ λႵѮᏘಒझ(RAW 264.7 cell line)ǵΓᜪ‫ޤ‬೽΢Ҝಒझਲ਼(A549 cell line)ϷΓᜪ՟ѮᏘಒझ(THP-1 cell line)ᖼԾ American Type. 34.

(45) Culture Colection(RockvlleǴMD)ǶDMEMǴpenicillinǴstreptomycin Ϸ fetal bovine serum ᖼԾ Gibco LifeTechnologies (GaithersburgǴ MD)Ƕ RAW264.7 Ϸ A549 ಒझ୻Ꭶӧ Dulbecco's modified Eagle's mediurm (DMEM)ǴԶ THP-1 ಒझ୻Ꭶӧ RPMI 1640 ύǴ‫୻ޣٿ‬Ꭶ నύ֖Ԗ 10% FBSǵpenicillin (100 unit/ml)аϷ streptomycin (100 ȝg/ml)ύǴ࿼‫ ܭ‬37ʚ֖Ԗ 5%Β਼ϯᅹಒझ୻ᎦጃύǶԶҭᖼԾ American Type Culture Collection (RockvlleǴMD)Ƕ ಒझ(5 x104 cells/well)ᅿ‫ ܭ‬96-Wellplates୻ᎦǶаPBSؑ ࢱǶӃуΕύᛰࡕ 1 λਔǴӆуΕLPS (1 ȝg/ml)‫ڈ‬ᐟಒझǴ႖‫୻ڹ‬ ᎦǶаphosphate-bufiered salinc (PBS)ؑࢱࡕǴᛰ‫ނ‬բҔ 24 λਔǶ уΕMTT(3-(4, 5-dimethylthiazolyl-2)-2, 5-diphenyltetrazolium bromide)Ǵ4 λਔǴӆуΕ 20% TritonX.100 (final 0.5%)ྋှಒझǶ ஒ 96-well plates‫ܫ‬Εspectra MAX 340 (Molecular DevicesǴ SunnyvaleǴCalifomia,USA)Ǵа 570 nmٰෳໆ֎ӀॶᡂϯǶ. 9ǵሌᙋණ‫ܭ‬ଯਏన࣬ቫ‫݋‬ϐԋҽϩ‫(݋‬High Performance Liquid Chromatography) ଞჹቷϣሌᙋණȐ501 ‫ץ‬ȑύ Chlorogenic acidǵLuteolinǵacrtiin Ϸ Hesperidin բ‫ۓ‬ໆϩ‫݋‬Ƕаଯਏన࣬ቫ‫ݤ݋‬຾Չ᠘ձϷ֖ໆෳ‫ۓ‬Ǵ ଯਏన࣬ቫ‫݋‬ሺǺHitachi D-7000 Interface , L-7100 pump , L-7455 DAD , L-7200AutosamplerǶଯਏన࣬ቫ‫݋‬చҹǺቫ‫݋‬ᆅࢊǺMightysil RP-18ǴGP 250×4.6 mm (5͔m)Ǵ౽୏࣬Ǻ. 35.

(46) Table 3.2. The time and partition of solvent Time(min). Acetonitrile(%). 0.3 %H3PO4 (%). 0. 10. 90. 45. 25. 75. 55. 70. 30. 60. 10. 90. 65. 10. 90. ࢬೲǺ1 ml/ϩដ! ᔠෳ‫ߏݢ‬Ǻ350 nm! ‫ݙ‬ΕໆǺ10 ͔l ኱ྗࠔଛᇙǺ‫ڗ‬аଛᇙϐ Chlorogenic acidǵLuteolinǵacrtiin Ϸ Hesperidin ኱ྗྋనǶᔠࠔྋనϐፓᇙǺ‫ڗ‬ኬࠔΟԛӚऊ 0.5 g Ѱ ѓǴуΕ 10 ml ‫ ޑ‬70 % MeOHǶ࿼‫ܭ‬଑ᙯԄ᎜ᕏᐒǴ200 rpmǴ60 ϩដ๧‫ڗ‬Ǵၸᘠࡕӆа 70 % MeOH ‫ۓ‬৒Կ 10 mlǴӆϩձ‫ ڗ‬1 ml ‫ۓ‬৒Կ 5 mlǴ‫ڗ‬፾ໆ೯ၸ 0.45 ͔m ᘠጢ‫ٮ‬բᔠనǶ 10ǵUIQ.2 ൂਡಒझჹ!B65: ΢Ҝಒझ‫ߕޑ‬๱Κϩ‫݋‬ ߕ๱ჴᡍЬाࢂҔhuman monocytic cell line THP-1 ჹ΢Ҝಒ झԖᛙ‫ߕޑۓ‬๱ΚౢғǴTHP-1 cells ୻ᎦӧRPMI containing 1% FBS Ϸ 5 ȝg/ml calcein-AM (Sigma Chemicals, St. Louis, MO) ӧ 37ºC Π 30 ϩដǴᑻӀ኱‫ޑۓ‬THP-1ȐFluorescence-labeled cellsȑ ፓԋ 1×106 cells/mlǹќѦஒൂቫ‫ޑ‬A549 cellȐmonolayerȑ୻Ꭶӧ 6-well plates ύуΕᐈҜન˞λਔǴϷTNFĮ ‫ ܈‬IL-1ȕǴ4 λਔǴஒ A549 аPBSࢱΒԛӆуΕ 100 ȝl (1×105) labeled THP-1 cells/well (4:1 ratio of monocytes to epithelial cells)Ǵ‫ ܭ‬37 ºCΠϸᔈ 30 ϩ ដǶӆҔPBSᇸᇸࢱΟԛǶӆஒcells‫ܫ‬ӧᑻӀᡉ༾᜔ȐAxiovert S100 microscopeǴZeiss, Jena, GermanyȑΠྣ࣬Ϸीኧ೏኱‫ޑۓ‬ಒझኧǶ. 36.

(47) 11ǵࢬԄಒझሺෳ‫ߕۓ‬๱ϩη ஒ 5 × 105 ঁA549 ಒझ୻Ꭶӧ 6 well plates΋ϺࡕǴҔPBSࢱ ΒԛǴӆႣӃ๏ϒᛰ‫ނ‬΋λਔǴӆ‫׫‬ᆶ 40 ng/ml TNFĮ ‫ ܈‬10 ng/ml IL-1ȕ୻ᎦѤλਔࡕǴҔPBSࢱΒԛǴӆҔtrypsin solution (Life Technologies)ϪΠಒझǴҔόӕᑻӀ኱‫ޑۓ‬VCAM-1, ICAM-1 ‫ל‬ᡏ (Becton Dickinson, Heidelberg, Germany)ᆶA549 ಒझբҔ΋λ ਔǴӆҔҔPBSࢱΒԛǴനࡕа 300Ȝ ‫ޑ‬PBSѺණಒझǴӆаFACScan cytometer (Becton Dickinson)ϩ‫݋‬Ƕ‫؂‬ԛԏΕ 10,000 ᗭಒझǴ଺Ο ख़ፄǶ 12ǵ಍ीБ‫ݤ‬ ճҔStudent’s t-testբ಍ीϩ‫݋‬ǴКၨӚಔ໔ϐৡ౦Ƕჴᡍ่݀ аmeans²SEM߄ҢǴpॶλ‫ܭ‬0.05ຎࣁ‫ڀ‬಍ी΢‫ޑ‬ᡉ๱‫܄‬Ƕ. 37.

(48) ಃѤക ่. ݀. ಃ΋೽ҽ ሌᙋණჹLPSᇨว೷ԋ‫ޤ‬ว‫ݹ‬ԴႵኳԄߥៈբҔϷխࣝፓ࿯ ᐒ‫ڋ‬ϩ‫݋‬ ಃ΋࿯ όӕᐚࡋLPS೷ԋ‫ޤ‬೽ཞ໾‫ޑ‬ቹៜ (the morphologic damage of lungs exposed to LPS)! ӧკ΋ᡉҢǴόӕᐚࡋ‫ޑ‬LPS༙ΕႵ‫܌ࡕޤ‬Їଆ‫ޑ‬ಔᙃว ‫ޑݹ‬ำࡋǴҗ‫ޤ‬ಔᙃϪТٰ࣮ǴႵ‫ډڙޤ‬LPS‫װ‬ᔐ24λਔࡕǴё࣮ ‫ډ‬ᒿLPSᐚࡋቚуԶ‫ޤ‬ཞ໾‫ރޑ‬ᄊΨቚуǴ྽Ⴕ‫ډڙޤ‬LPS‫װ‬ᔐౢ ғ‫ޤ‬೽ว‫ݹ‬ϸᔈǴ‫ॺך‬ёаᢀჸว‫ޤޑݹ‬೽ಔᙃ཮ౢғ‫ޑ‬ᡂϯԖ(1) ‫ݰޤ‬Ꮫ‫ޑ‬ᡂࠆǵ(2)ӧ‫ޤ‬೽‫ݰޤޑ‬ϣԖрՈǵ(3)ӧ‫ݰޤ‬ϣ‫ډـ‬εໆ‫ޑ‬ ൂਡ‫܄‬қՈౚϷӭਡ‫܄‬қՈౚ੆ዎ(ӵკ4.1)ǶӧLPS 100~200 ȝg/kg ჹᇨว‫ݹޤ‬Ⴕё‫ډـ‬ၨᝄख़‫ޤޑ‬೽ว‫ݹ‬ǴՠӧLPS 200 ȝg/kg ‫ޑ‬ᇨว‫ݹޤ‬Ⴕ཮೷ԋႵଫԝΫԶԏό‫ډ‬ၗ਑Ǵࡺ‫ॺך‬ᒧҔΑLPS 100 ȝg/kgࣁჴᡍ٬Ҕ‫ޑ‬ᐚࡋǶ ಃΒ࿯ ሌᙋණჹLPS೷ԋ‫ޤ‬೽ཞ໾‫ޑ‬ቹៜ (The effect of GGS on the morphologic damage of lungs exposed to LPS) ҅த‫ޤޑ‬೽ಔᙃϪТᔈё‫ֹډـ‬᏾‫ݰޤޑ‬ᏛϷଳృ‫ݰޤޑ‬ ‫ޜ‬໔(ӵკ4.2A)Ǵ྽Ⴕ‫ډڙޤ‬LPS‫װ‬ᔐౢғΑ‫ޤ‬೽ว‫ݹ‬ϸᔈǴ‫ॺך‬ ёаᢀჸว‫ޤޑݹ‬೽ಔᙃ‫(ډـ‬1)‫ݰޤ‬Ꮫ‫ޑ‬ᡂࠆǵ(2)ӧ‫ޤ‬೽‫ݰޤޑ‬ϣ ԖрՈǵ(3)ӧ‫ޑݰޤ‬ϣ‫ॺך‬ё‫ډـ‬εໆ‫ൂޑ‬ਡ‫܄‬қՈౚϷӭਡ‫܄‬қ Ոౚ੆ዎ(ӵკ4.2B)Ƕ‫ॺך‬٬ҔΑόӕᐚࡋ‫ޑ‬ሌᙋණ(0.25 mg/kg (C), 0.5 mg/kg (D), 1 mg/kg (E), and 2 mg/kg (F))ᗯ१ԴႵϷӕਔ ਻ᆅϣ‫׫‬ᆶLPS ‫ޑ‬24λਔࡕǴ‫ॺך‬ёа࣮‫ޤډ‬೽ಔᙃ‫ډڙ‬LPS‫װ‬ᔐ ࡕǴᒿ๱ሌᙋණ٬Ҕᐚࡋ‫ޑ‬ቚуԶԖܴᡉ‫ׯޑ‬๓Ǵӵ‫ݰޤ‬Ꮫ‫ޑ‬ᡂᖓǵ ӧ‫ݰޤ‬ϣрՈ෧ϿϷӧ‫ݰޤ‬ϣ੆ዎൂਡ‫܄‬қՈౚϷӭਡ‫܄‬қՈౚΨ 38.

(49) ܴᡉ෧ϿǴё‫ـ‬٬ҔሌᙋණჹLPS೷ԋ‫ޤ‬೽ཞ໾Ԗ‫ݯ‬ᕍ‫ޑ‬ԋਏǶ. Figure 4.1. the morphologic damage of lungs exposed to different concentration of LPS. Representative histologic sections of lungs harvested 24 hours after the intratracheal instillation of LPS. Magnification Ø 100.. 39.

(50) Figure. 4.2. The lung morphology changes of GGS treatment. Representative histologic sections of lungs harvested 24 hours after the intratracheal instillation of LPS (B), or none (A) in mice, and oral administration of 0.25 mg/kg of GGS (C), 0.5 mg/kg of GGS (D), 1 mg/kg of GGS (E), and 2 mg/kg of GGS (F) in LPS-induced lung inflammatory mice. Pictures are representative of n =6 mice/treatment group. Magnification Ø. 200.. 40.

(51) Figure 4.3. The lung morphology changes of GGS, HBX and Hesperidin treatment. Representative histologic sections of lungs harvested 24 hours after the intratracheal instillation of LPS (A), in mice, and oral administration of 1 mg/kg of GGS (B), 1 mg/kg of HBX (C), and. 200 mg/kg of Hesperidin (D) in LPS-induced lung inflammatory mice. Magnification Ø. 200.. 41.

(52) ‫ॺך‬Ψஒሌᙋණᆶќ΋ᅿ‫ל‬ว‫ૈݹ‬ΚКၨ১‫ޑ‬ύᛰБᘞқණ(Xia-Bai-san; XBS)ǴаϷӧሌᙋණύ֖Ԗ‫ל‬ว‫ૈݹ‬ Κ໳✉ᜪ-ᐈҜન(Hesperidin; HES)຾ՉಔᙃϪТ΢‫ޑ‬К ၨǴܴᡉ‫ޑ‬ሌᙋණ‫ݯ‬ᕍಔᆶᐈҜન‫ݯ‬ᕍಔӧ‫ݯ‬ᕍ24λਔࡕਏ ݀КၨӳǴԶᘞқණಔΨԖ‫ׯ‬๓ว‫ރݹ‬ᄊǴՠਏ݀ۘόϷ΢ ‫ٿ‬ᅿᛰȐӵკ4.3ȑ Ƕ. ಃΟ࿯ ሌᙋණჹLPS೷ԋ‫ޤ‬೽ཞ໾ౢғ‫਻ޑ‬ᆅϣᅖрనϐቹៜ (The effect of GGS on the formation of high-permeability edema after intratracheal instillation of LPS) ARDS‫ޑ‬஻‫ޤޣ‬೽཮ౢғεໆ‫ޑ‬ᅖрన೷ԋ‫ޤ‬೽Н ဍǴߔᛖΑ‫ޤ‬೽‫ޑ‬ඤ਻բҔԶᏤठੰ஻ԝΫǴ‫܌‬а‫ׯ‬๓‫ޤ‬೽ ᅖрన‫ޑ‬ໆϷᅖрనϣೈқᐚࡋࢂ΋ঁࡐख़ा‫ࡰޑ‬኱Ƕ‫ॺך‬ ෳໆΑ‫ޑޤ‬ख़ໆϷ‫ࢱؑݰޤ‬నϣ‫ޑ‬ೈқ፦‫ޑ‬ᐚࡋٰ߄౜‫ޤ‬ ೽ᅖрన‫ޑ‬ӭჲǶӵკ4.4AᡉҢǴ҅தԴႵ‫ޤޑ‬ख़ऊ0.15 ± 0.027 gǴԶLPSᇨวႵ‫ޤ‬ว‫ݹ‬24λਔࡕǴख़ऊ0.24 ± 0.054 gǴ ‫ޤ‬೽ख़ໆቚуε‫ܭ‬60%Ǵё‫ޤـ‬೽ϣԖ೚‫ޑ‬ᅖрన‫ޑ‬ౢ ғǶՠӧӕਔᗯ१όӕᐚࡋ‫ޑ‬ሌᙋණ24λਔࡕǴ‫ޤځ‬೽‫ޑ‬ख़ ໆ෧ࣁ0.22 ± 0.035 g (0.5 mg/kg ሌᙋණ)ǵ0.18 ± 0.022 g (1 mg/kgሌᙋණ)ǵ0.17 ± 0.012 g (2 mg/kg ሌᙋණ)Ǵ‫ޤ‬೽ ख़ໆԖܴᡉᒿ๱‫׫‬ᆶ‫ޑ‬ሌᙋණᐚࡋቚуԖ‫܌‬Πफ़Ƕӧ‫ؑݰޤ‬ ࢱనϣೈқ፦ᐚࡋ΢Ǵӵკ4.40BᡉҢǴ҅தԴႵ‫ؑݰޤޑ‬ ࢱనϣೈқ፦ᐚࡋࣁ0.21 ± 0.071 ȝg/mlǴԶ‫ډڙ‬LPS‫װ‬ᔐ24 λ ਔ ࡕ ‫ ࢱ ؑ ݰ ޤ‬న ϣ ೈ қ ፦ ᐚ ࡋ ቚ у ࣁ 0.54 ± 0.16 ȝg/mlǴՠӧӕਔᗯ१όӕᐚࡋ‫ޑ‬ሌᙋණ24λਔࡕǴё‫ډـ‬ ‫ࢱؑݰޤ‬నϣೈқ፦ᐚࡋԖܴᡉ‫ޑ‬όӕำࡋ‫ޑ‬Πफ़Ǵ0.50 ± 0.04 ȝg/ml (0.5 mg/kg ሌᙋණ)ǵ0.46 ± 0.034 ȝg/ml (1 mg/kg ሌᙋණ)Ϸ 0.43 ± 0.026 ȝg/ml (2 mg/kg ሌᙋණ)Ƕ 42.

(53) ӧ‫ޤ‬೽ख़ໆϷ‫ࢱؑݰޤ‬నϣೈқ፦ᐚࡋ‫ݯޑ‬ᕍٰ่࣮݀Ǵሌ ᙋණёа෧ϿLPS‫װ‬ᔐ‫ޤ‬೽೷ԋεໆ‫ޑ‬ᅖрన‫ރޑ‬ᄊǴԶቚ уARDS‫ޤ܈‬೽࡚‫܄‬ว‫ݹ‬஻‫ޣ‬Ӹࢲ‫ޑ‬ᐒ཮Ƕ ‫ॺך‬ΨӕਔКၨሌᙋණǵᘞқණϷᐈҜનჹLPSᇨว Ⴕ‫ޤ‬ว‫ޤޑݹ‬ಔᙃख़ໆϷ‫ࢱؑݰޤ‬నϣೈқ፦ᐚࡋȐӵკ 4.5ȑ Ǵਏ݀നӳ‫ࢂޑ‬ᐈҜનǴࣗԛࣁሌᙋණǴԶᘞқණᗨฅ К߻‫ޣٿ‬ৡՠΨԖᕍਏǶ. ಃѤ࿯ ሌᙋණჹLPS೷ԋ‫ޤ‬೽ཞ໾ౢғ‫ࢱؑݰޤޑ‬నύ‫ޑ‬Ոౚໆ ϐ ቹ ៜ (The effect of GGS on the lung leukocyte accumulation after intratracheal instillation of LPS) ྽‫ޤ‬೽҅ӧ຾Չว‫ݹ‬ϸᔈਔǴ‫ޑݰޤ‬ϣ೽‫ډـ‬εໆ‫ൂޑ‬ ਡ‫܄‬қՈౚϷӭਡ‫܄‬қՈౚ੆ዎǴ‫܌‬а྽җ਻ᆅύ‫ڗ‬ள‫ݰޤ‬ ؑࢱనਔǴӧ‫ࢱؑݰޤ‬నύߡ཮Ԗ೚ӭՈౚӸӧǴԶЪኧໆ ཮࡚ೲ‫ޑ‬ϲଯǴ‫ࢱؑݰޤ‬నύ‫ޑ‬Ոౚ֖ԖѮᏘಒझǵқՈౚ ϷరЃౚǴӧ࡚‫܄‬ว‫ݹ‬ϸᔈύനख़ा‫ޑ‬қՈౚࢂύ‫܄‬қՈ ౚǴ೭ԜՈౚ཮ϩ‫ݜ‬೚ӭߦ຾ว‫ݹ‬ϸᔈ࡭ុ຾Չ‫ޑ‬ว‫ݹ‬Ӣ ηǴ‫܌‬а෧Ͽ‫ࢱؑݰޤ‬నύՈౚኧໆΨࢂफ़եว‫ݹ‬ϸᔈख़ा ‫ޑ‬πբϐ΋Ƕӧკ4.6ύᡉҢ๱Ǵ҅த‫ޑ‬Ⴕ‫ޤ‬ύ‫ࢱؑݰޤޑ‬ నύѝԖϿໆ‫ޑ‬ՈౚӸӧ(total cells: 10.0 ± 2.82 × 104 cells/BALF)ǴϷཱུϿኧ‫ޑ‬ύ‫܄‬қՈౚ‫ޑ‬Ӹӧ(0.9 ± 0.61 × 104 cells/BALF)Ǵӧ਻ᆅϣ‫׫‬ᆶLPSٰᇨว‫ޤ‬೽ว‫ݹ‬ϸᔈ24 λਔࡕǴё‫ډـ‬εໆ‫ޑ‬Ոౚ೏֎Ї‫ޤډ‬ύǴӸӧ‫ࢱؑݰޤ‬న ύ‫ޑ‬Ոౚኧߡӛ΢ᚹϲȐtotal cellsǺ89.3 ± 7.4 × 104 cells/BALFȑ ǴЀ‫ࢂځ‬ύ‫܄‬қՈౚ‫ࢂ׳‬՞‫ޤ‬೽‫ޑ‬ಒझύ೽ϩॶ Ȑ71.5 ± 1.34 × 104 cells/BALF at 24 hoursȑǴསҢ๱‫ޤ‬೽ ว‫ݹ‬ϸᔈ҅ӧࢵ෢‫ޑ‬຾ՉύǴՠࢂ྽‫ॺך‬ӕਔᗯ१όӕᐚࡋ ‫ޑ‬ሌᙋණࡕǴ‫ࢱؑݰޤ‬నύ‫ޑ‬ՈౚኧߡԖᡉ๱‫ޑ‬෧ϿǴ 43.

(54) GGS-treated GGS-tre dose GGS-treated ateddose dose (mg/Kg) (mg/Kg) (mg/Kg) Figure 4.4. The weight of lung and concentrations of proteins in BALF(I). BALB/c mice received an intratracheal instillation of lipopolysaccharide and were treated with different doses of GGS (n = 6) or naïve mice received PBS instead of LPS intratracheally (n = 6), and oral administration of different concentration of GGS in LPS-induced lung inflammatory mice. After 24 hours, mice sacrifice and measure the weight of lung (A) and proteins of BALF (B) as an index of high-permeability pulmonary edema. Means ± SEM. *P < 0.05 LPS-treated vs. Naïve. **P < 0.05 GGS vs. LPS-treated mice.. 44.

(55) Figure 4.5. The weight of lung and concentrations of proteins in BALF(II). BALB/c mice received an intratracheal instillation of lipopolysaccharide and were treated with GGS, XBS, and HES or naïve mice received PBS instead of LPS intratracheally. After. 4 hours and. 24 hours, mice sacrifice and measure the. weight of lung (A) and proteins of BALF (B) as an index of high-permeability pulmonary edema.. 45.

(56) Figure 4.6. The cell counts in the BALF. BALB/c mice received an intratracheal instillation of lipopolysaccharide or not (naïve) and were treated with different doses of GGS. Total cell counts Ű and Neutrophil cell counts ɍ of BALF were determined at 24 hours after LPS. Means ± SEM. **P < 0.05 LPS vs. Naïve, *P< 0.05 GGS vs. LPS treated mice.. 46.

(57) ӧᕴՈౚኧ΢෧ࣁ76.2 ± 2.16 × 104 cells/BALF (0.5 mg/kg ሌᙋණ)ǵ47.3 ± 8.91 × 104 cells/BALF (1 mg/kg ሌᙋණ) Ϸ40.5 ± 4.72× 104 cells/BALF (2 mg/kg ሌᙋණ)ǹԶύ‫܄‬ қՈౚ‫ޑ‬ኧໆ‫ܴࢂ׳‬ᡉ‫ޑ‬फ़եǴӵ64.5± 1.23 × 104 cells/BALFȐ0.5 mg/kg ሌᙋණȑǵ 40.5 ± 0.67 × 104 cells/BALFȐ1 mg/kg ሌᙋණȑϷ 22.6 ± 0.51 × 104 cells/BALFȐ2 mg/kg ሌᙋණȑ Ƕ ሌᙋණǵᘞқණϷᐈҜનჹLPSᇨวႵ‫ޤ‬ว‫ޑݹ‬ᕴಒझ ኧ‫ޑ‬Кၨ΢ǴΟ‫ࣣޣ‬ԖਏǴԶᐈҜનಔᓬ‫ܭ‬ሌᙋණಔǵᘞқ ණಔȐკ4.7Aȑ Ǵՠӧύ‫܄‬қՈౚ‫ޑ‬ኧໆ΢Ǵࠅࢂሌᙋණಔǵ ᘞқණಔᓬ‫ܭ‬ᐈҜનಔȐკ4.7BȑǶ ΢ॊ่݀ගҢ‫ॺך‬๏ᆶሌᙋණǵᘞқණϷᐈҜનёа෧ Ͽ‫ޤ‬೽ว‫ݹ‬ύ‫ޑ‬Ոౚ੆ዎǴ೭ᆶ೚ӭᇨ٬Ոౚ຾Ε‫ޤ‬೽‫ޑ‬ಒ झᐟનԖᜢǴ‫܌‬ааΠ‫ޑ‬ჴᡍߡ௖૸ᆶ‫ޤ‬೽ว‫࣬ݹ‬ᜢ‫ޑ‬ว‫ݹ‬ ӢηǶ ಃϖ࿯! ሌᙋණჹLPSᇨว‫ޑ‬ᖿϯӢη‫ޑ‬ቹៜ(The effect of GGS on the expression of chemokines in lungs exposed to LPS) ӧ‫ޤ‬೽ว‫ݹ‬ϸᔈύёаᇨ٬εໆՈౚ຾Ε‫ޜݰޤ‬໔ϣ ‫ޑ‬ᖿϯӢηԖǺIL-8(Γᜪ)ǵKC(ԴႵ)ǵMCP-1ϷMIP-2Ƕ‫ך‬ ॺёаӧკ4.8࣮‫ډ‬ԴႵύԖᜢ‫ޑ‬ᖿϯӢηᆶሌᙋණ‫ޑ‬ᜢ ߯Ǵӧკ4.8˽ύǴ҅தԴႵ‫ࢱؑݰޤ‬నύ‫ޑ‬KCࢂ࣬྽ե‫ޑ‬ ໆ(35.78 ± 5.23 pg/ml)Ǵ྽ӧ਻ᆅϣ‫׫‬ᆶLPSٰᇨว‫ޤ‬೽‫ޑ‬ ว ‫ ݹ‬ϸ ᔈ 24 λ ਔ ࡕ Ǵ KC ‫ ޑ‬ໆ ࡚ ቃ ቚ у (379.78 ± 47.65 pg/ml) Ǵ྽‫ॺך‬ӕਔаα‫׫‬ᆶ1 mg/kg‫ޑ‬ሌᙋණࡕǴ‫ؑݰޤ‬ ࢱనύ‫ޑ‬KCԖཀက‫ޑ‬Πफ़‫ډ‬295.81±4.41 pg/mlǶ ‫ॺך‬Ψӕਔ‫ڗ‬Π‫ޤ‬೽ಔᙃǴ‫ځڗܜ‬mRNAǴаRT-PCR ‫ޑ‬БԄǴჹќѦΒᅿᖿϯӢηȐMCP-1 ϷMIP-2ȑ຾ՉଓᙫǴ ӧკ4.8˾Ǵ‫ډـ‬MCP-1ϷMIP-2ӧ҅தԴႵࢂό߄౜‫ޑ‬Ǵՠ 47.

(58) Figure 4.7. The tatol cell counts and neutrophil number in the BALF. BALB/c. mice. received. an. intratracheal. instillation. of. lipopolysaccharide or not (naïve) and were treated with GGS, XBS, and HES.. 4 hours and. 24 hours after LPS.. 48.