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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√– ß§å‡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 “√ °—¥
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