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Journal of Thai Traditional & Alternative Medicine Vol. 8 Nos. 2-3 May-December 2010 ÒÙ˜
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∑’Ëπà“ π„®¡“°¡“¬ „πªí®®ÿ∫—π¡’À≈“¬√“¬ß“π°“√«‘®—¬‡°’ˬ«°—∫ ”À√—∫‡μ√’¬¡ “√ °—¥®“°‡Àßâ“¢à“
¡ÿπ‰æ√∑’Ë¡’ƒ∑∏‘Ϭ—∫¬—Èß°“√∑”ß“π¢Õ߇Õπ‰´¡å pancreatic Ò.Ò.ı °“√§—ÈππÈ” ”À√—∫‡μ√’¬¡ “√ °—¥®“°
Ú-Ù
lipase ƒ∑∏‘Ϭ—∫¬—Èß°“√∑”ß“π¢Õ߇Õπ‰´¡å HMG-CoA re- º≈μ–≈‘ߪ≈‘ß ¥
ı-˜
ductase „π¢≥–∑’˃∑∏‘Ϭ—∫¬—Èß°“√¥Ÿ¥´÷¡§Õ‡≈ ‡μÕ√Õ≈®“° Ò.Ú °“√∑”·Àâß “√ °—¥ “√ °—¥‡Õ∑“πÕ≈®“°
¯
¡ÿπ‰æ√‰∑¬¬—ß¡’°“√»÷°…“‰¡à¡“°π—° ¥—ßπ—Èπ °“√»÷°…“π’È®÷ß ¡ÿπ‰æ√™π‘¥μà“ß Ê ∑”·Àâß‚¥¬«‘∏’√–‡À¬¿“¬„μâ ÿ≠≠“°“»
¡’«—μ∂ÿª√– ߧå‡æ◊ËÕ»÷°…“ƒ∑∏‘Ϭ—∫¬—Èß°“√¥Ÿ¥´÷¡§Õ‡≈ ‡μÕ√Õ≈ ¥â«¬‡§√◊ËÕß Eyela NE rotary evaporator “√ °—¥πÈ”∑”
(cholesterol uptake inhibition) ‡¢â“ Ÿà‡´≈≈å Caco-2 ¢Õß ·Àâß‚¥¬«‘∏’ freeze dry à«π “√ °—¥∑’ˉ¥â®“°«‘∏’ SFE ‰¡à

“√ °—¥ ¡ÿπ‰æ√‰∑¬ ‡æ◊ËÕ à߇ √‘¡»—°¬¿“æ¢Õß ¡ÿπ‰æ√‰∑¬ μâÕߺà“π°√–∫«π°“√∑”„Àâ·Àâߢâ“ßμâπ
„À⇪ìπ∑“߇≈◊Õ°„π°“√√—°…“À√◊ÕªÑÕß°—π°“√‡°‘¥¿“«–‰¢¡—π„π Ò.Û °“√‡μ√’¬¡ “√ °—¥‡æ◊ËÕ∑¥ Õ∫ƒ∑∏‘Ϭ—∫¬—Èß°“√

‡≈◊Õ¥ ŸßμàÕ‰ª„πÕπ“§μ ¥Ÿ¥´÷¡§Õ‡≈ ‡μÕ√Õ≈ “√ °—¥ ¡ÿπ‰æ√∑’ˉ¡à “¡“√∂≈–≈“¬
πÈ”‰¥â μâÕ߇μ√’¬¡„ÀâÕ¬Ÿà„π√Ÿª¢Õß “√ª√–°Õ∫‡™‘ß´âÕπ°—∫
√–‡∫’¬∫«‘∏’»÷°…“
polyvinylpyrrolidone (PVP) 40,000 (Fluka, United State)
¡ÿπ‰æ√
‚¥¬ “√ °—¥ ¡ÿπ‰æ√·μà≈–™π‘¥®–¡’Õ—μ√“ à«π∑’ˇÀ¡“–„π°“√
‡°Á∫μ—«Õ¬à“ß ¡ÿπ‰æ√ ®”π«π ÒÒ ™π‘¥ ‰¥â·°à „∫∫—«À≈«ß ‡°‘¥ “√ª√–°Õ∫‡™‘ß´âÕπ∑’Ë·μ°μà“ß°—π

„∫™–¡«ß „∫μ–≈‘ߪ≈‘ß º≈μ–≈‘ߪ≈‘ß „∫¡–√ÿ¡ „∫¬Õ „∫Ω√—Ëß Ú. °“√»÷°…“§«“¡‡ªìπæ‘…μàÕ‡´≈≈å Caco-2
‡Àßâ“¢à“ à«π„μ⥑πªí≠®¢—π∏å „∫¡–°√Ÿ¥ ·≈–„∫¬à“π“ß „™â«‘∏’ MTT assay ‡æ◊ËÕ»÷°…“º≈¢Õß “√ °—¥ ¡ÿπ‰æ√
˘
π” ¡ÿπ‰æ√ ¬°‡«âπ„∫¡–°√Ÿ¥ ·≈–º≈μ–≈‘ߪ≈‘ß ≈â“ß„Àâ ‰∑¬μàÕ cell viability ¢Õ߇´≈≈å Caco-2
–Õ“¥ À—Ëπ‡ªìπ™‘Èπ‡≈Á° Ê ·≈â«∑”„Àâ·Àâß∑’ËÕÿ≥À¿Ÿ¡‘ª√–¡“≥ Ùı- Ú.Ò °“√‡æ“–‡≈’Ȭ߇´≈≈å Caco-2
ı˚´ ®“°π—Èππ”¡“∫¥‡ªìπºß≈–‡Õ’¬¥ ‡≈’Ȭ߇´≈≈å Caco-2 ¥â«¬Õ“À“√‡≈’Ȭ߇´≈≈å DMEM/
à«π„∫¡–°√Ÿ¥·≈–º≈μ–≈‘ߪ≈‘ß ‡¡◊ËÕ≈â“ß„Àâ –Õ“¥·≈â« F12 ∑’Ë¡’ à«πº ¡¢Õß Ò% FBS ·≈– Ò% penicillin-strep-

𔉪‡μ√’¬¡ “√ °—¥ ¡ÿπ‰æ√‚¥¬‰¡àμâÕߺà“π°√–∫«π°“√ tomycin ∫à¡„π incubator ∑’ËÕÿ≥À¿Ÿ¡‘ Û˜˚´ ·≈– CO ı%
2
∑”„Àâ·Àâß ‡¡◊ËÕ‡´≈≈å Caco-2 ‡®√‘≠‡μ‘∫‚μ∑—Ë«®“π‡≈’È¬ß ∑”°“√ subcul-
ture ∑ÿ° Û-Ù «—π ‚¥¬ detach ‡´≈≈å¥â«¬ .Úı% trypsin
«‘∏’°“√»÷°…“ 2+ 2+
„π Ca -, Mg -free phosphate buffer (PBS) ∑’Ë¡’ EDTA
Ò. °“√‡μ√’¬¡ “√ °—¥ ¡ÿπ‰æ√ .Ú °./≈. ‡ªìπ à«πª√–°Õ∫

Ò.Ò °“√ °—¥ “√®“° ¡ÿπ‰æ√ „π°“√«‘®—¬π’È¡’«‘∏’ Ú.Ú Cell viability test (MTT assay)
°“√‡μ√’¬¡ “√ °—¥®“° ¡ÿπ‰æ√ ı «‘∏’ §◊Õ ‡≈’Ȭ߇´≈≈å Caco-2 „π ˘ˆ well plate ∑’˧«“¡Àπ“
Ò.Ò.Ò °“√À¡—° ”À√—∫‡μ√’¬¡ “√ °—¥ ·πàπ Ò, cells/well ‚¥¬∑”°“√‡æ“–‡≈’Ȭ߄π CO incu-
2
‡Õ∑“πÕ≈®“°„∫¡–°√Ÿ¥ ¥ bator ‡ªìπ‡«≈“ Ò «—π ®“°π—Èππ”‡´≈≈å Caco-2 ¡“‡≈’Ȭ߄π
Ò.Ò.Ú °“√ °—¥·∫∫μàÕ‡π◊ËÕß ‚¥¬„™â soxhlet Õ“À“√‡≈’Ȭ߇´≈≈å„π ¿“«–∑’Ë¡’·≈–‰¡à¡’ “√ °—¥ ¡ÿπ‰æ√∑’Ë

apparatus ”À√—∫‡μ√’¬¡ “√ °—¥‡Õ∑“πÕ≈ ®“°„∫∫—«À≈«ß §«“¡‡¢â¡¢âπμà“ß Ê º ¡Õ¬Ÿà ‡ªìπ‡«≈“ Ò «—π ‚¥¬„π™—Ë«‚¡ß∑’Ë
„∫™–¡«ß „∫μ–≈‘ߪ≈‘ß „∫¡–√ÿ¡ „∫¬Õ „∫Ω√—Ëß ªí≠®¢—π∏å ÚÚ ‡μ‘¡ “√≈–≈“¬ .ı% MTT Ò ¡§≈. ≈߉ª„π medium
·≈–„∫¬à“π“ß ·≈–‡¡◊ËÕ§√∫ ÚÙ ™—Ë«‚¡ß ¥Ÿ¥Õ“À“√‡≈’Ȭ߇´≈≈åÕÕ°„ÀâÀ¡¥

Ò.Ò.Û °“√μâ¡ °—¥¥â«¬πÈ” (reflux) ”À√—∫ ·≈â«‡μ‘¡ “√≈–≈“¬ DMSO-EtOH (Ò:Ò% v/v) ª√‘¡“μ√ Ú
‡μ√’¬¡ “√ °—¥πÈ”®“°„∫μ–≈‘ߪ≈‘ß·≈–„∫¡–√ÿ¡ μl/well ∫ࡉ«â∑’ËÕÿ≥À¿Ÿ¡‘ Û˜˚´ ‡ªìπ‡«≈“ Ò π“∑’ ·≈â«π”‰ª
Ò.Ò.Ù °“√ °—¥¥â«¬«‘∏’ supercritical fluid «—¥§à“°“√¥Ÿ¥°≈◊π· ß∑’˧«“¡¬“«§≈◊Ëπ ı˘ı π“‚π‡¡μ√

extraction (SFE) ‚¥¬‰¡à„™âμ—«∑”≈–≈“¬„¥ Ê ™à«¬„π°“√ °—¥ Û. °“√¬—∫¬—Èß°“√¥Ÿ¥´÷¡§Õ‡≈ ‡μÕ√Õ≈¢Õß “√ °—¥