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             Journal of Thai Traditional & Alternative Medicine                      Vol. 8 Nos. 2-3 May-December 2010 ÒıÛ
                          Û
             μâ“π‡™◊ÈÕ®ÿ≈‘π∑√’¬å ¬—∫¬—Èß°“√¥Ÿ¥´÷¡§Õ‡≈ ‡μÕ√Õ≈ ¬—∫¬—Èß  ·≈– Silica gel GF254 precoated plate, 0.25 mm
             ‡Õπ‰´¡å pancreatic lipase ·≈–¬—∫¬—È߇Õπ‰´¡å HMG CoA  (E. Merck, Germany)
                     Ù
             reductase ‡ªìπμâπ
                                                                                  «‘∏’°“√»÷°…“
                 °“√«‘®—¬π’È®÷ß¡’«—μ∂ÿª√– ß§å‡æ◊ËÕ»÷°…“ƒ∑∏‘Ï≈¥‰¢¡—π„π
             À≈Õ¥∑¥≈ÕߢÕß “√ °—¥®“°„∫™–¡«ß ‚¥¬»÷°…“º≈μàÕ°“√          Ò. °“√ °—¥ “√
             ¥Ÿ¥´÷¡§Õ‡≈ ‡μÕ√Õ≈‡¢â“ Ÿà‡´≈≈å≈”‰ â‡æ“–‡≈’È¬ß Caco-2 ´÷Ë߇ªìπ   °—¥„∫™–¡«ß¥â«¬ ˘ı% ‡Õ∑“πÕ≈ ¥â«¬«‘∏’ soxhlet
             cell-based assay ·≈–»÷°…“º≈°“√¬—∫¬—Èß°“√∑”ß“π¢Õß   extraction ·≈â«π” “√ °—¥∑’ˉ¥â‰ª√–‡À¬·Àâß¿“¬„μâ ÿ≠≠“°“»

             ‡Õπ‰´¡å pancreatic lipase ·≈– HMG Co-A reductase   ‰¥â “√ °—¥‡Õ∑“πÕ≈ (GC-1) ´÷Ëßπ”¡“ partition μàե⫬
             ´÷Ë߇ªìπ in vitro enzymatic assay ·≈–‰¥â∑”°“√∑¥ Õ∫  ‡Œ°‡´π ‰¥§≈Õ‚√¡’‡∑π·≈–∫‘«∑“πÕ≈ ·≈â«π” “√ °—¥·μà≈–

             ‡∫◊ÈÕßμâπ‡æ◊ËÕÀ“°≈ÿà¡ “√ ”§—≠·≈–»÷°…“∑“ß惰…‡§¡’‡æ◊ËÕÀ“   à«π∑’ˉ¥â‰ª√–‡À¬·Àâ߉¥â “√ °—¥‡Œ°‡´π (GC-2),  “√ °—¥
              “√ÕÕ°ƒ∑∏‘Ï≈¥‰¢¡—πÀ√◊Õ “√∫àß™’È (marker) „π°“√§«∫§ÿ¡  ‰¥§≈Õ‚√¡’‡∑π (GC-3),  “√ °—¥∫‘«∑“πÕ≈ (GC-4) ·≈– “√
             §ÿ≥¿“æ„∫™–¡«ßμàÕ‰ª„πÕπ“§μ ·≈–¬—߇ªìπ°“√‡æ‘Ë¡‡μ‘¡    °—¥πÈ” (GC-5) πÈ”Àπ—° ˆ˘.Ú, ¯ı.ˆ˜, Ò˘ˆ.˜ı ·≈–

             ¢âÕ¡Ÿ≈∑“ß惰…‡§¡’¢Õß„∫™–¡«ß´÷Ë߬—ß¡’√“¬ß“π‰¡à¡“°π—°  ÙÛ.˘ °√—¡ μ“¡≈”¥—∫ (§‘¥‡ªìπ√âÕ¬≈– Û.ˆÙ, Ù.ıÒ, Ò.Ûˆ
                                                                ·≈– Ú.Ú˜ μ“¡≈”¥—∫ ‡¡◊ËÕ‡∑’¬∫°—∫πÈ”Àπ—°„∫™–¡«ß·Àâß)
                            √–‡∫’¬∫«‘∏’»÷°…“                                                          ı
                                                                     Ú. °“√∑¥ Õ∫°≈ÿà¡ “√ ”§—≠„π„∫™–¡«ß
                  ¡ÿπ‰æ√                                             »÷°…“°≈ÿà¡¢Õß “√– ”§—≠∑’Ëæ∫„π “√ °—¥μà“ß Ê ®“°„∫
                 „∫™–¡«ß ¥ πÈ”Àπ—° Ò °‘‚≈°√—¡ π”¡“≈â“ßπÈ”„Àâ –Õ“¥  ™–¡«ß ‰¥â·°à terpenoids-steroids, flavonoids, phenolics,

             Õ∫∑’ËÕÿ≥À¿Ÿ¡‘ ı Õß»“‡´≈‡´’¬  ®π·Àâß π”‰ª∫¥À¬“∫‰¥â  alkaloids, anthraquinones, saponins ·≈– amines-amino
             ºß„∫™–¡«ß·Àâß πÈ”Àπ—° Ò.˘ °‘‚≈°√—¡ (§‘¥‡ªìπ√âÕ¬≈– Ò˘  acids ‚¥¬„™â phytochemical screening test
             ‡¡◊ËÕ‡∑’¬∫°—∫πÈ”Àπ—°„∫™–¡«ß ¥)                          Û. °“√‡μ√’¬¡ “√ °—¥‡æ◊ËÕ∑¥ Õ∫ƒ∑∏‘Ï≈¥‰¢¡—π„π

                 «— ¥ÿ·≈– “√‡§¡’                                À≈Õ¥∑¥≈Õß
                 μ—«∑”≈–≈“¬Õ‘π∑√’¬å∑’Ë„™â„π°“√‡μ√’¬¡ “√ °—¥ ¡ÿπ‰æ√    “√ °—¥ GC-1, GC-2, GC-3 ·≈– GC-4 ‰¡à “¡“√∂
             ·≈–·¬° “√„Àâ∫√‘ ÿ∑∏‘Ï„™â commercial grade ‚¥¬°≈—Ëπ´È”  ≈–≈“¬πÈ”‰¥â μâÕ߇μ√’¬¡„ÀâÕ¬Ÿà„π√Ÿª¢Õß “√ª√–°Õ∫‡™‘ß´âÕπ

             °àÕππ”¡“„™â                                        °—∫ polyvinylpyrrolidone 40,000 (PVP complex) ‚¥¬
                 DMSO-d  ·≈– CD OD (Wilmad Labglass, USA)        “√ °—¥·μà≈–™π‘¥®–¡’Õ—μ√“ à«π∑’ˇÀ¡“– ¡„π°“√‡°‘¥
                         6       3
              ”À√—∫°“√‡μ√’¬¡μ—«Õ¬à“߇æ◊ËÕ«‘‡§√“–Àå¥â«¬«‘∏’ NMR spec-   “√ª√–°Õ∫‡™‘ß´âÕπ∑’Ë·μ°μà“ß°—π  ”À√—∫ “√ °—¥ GC-5
             troscopy                                            “¡“√∂≈–≈“¬πÈ”‰¥â ®÷߉¡àμâÕ߇μ√’¬¡‡ªìπ PVP complex
                  “√‡§¡’Õ◊Ëπ Ê ‰¥â·°à anisaldehyde (Fluka, Germany)  Ù. °“√»÷°…“§«“¡‡ªìπæ‘…μàÕ‡´≈≈å Caco-2

             ·≈– sulfuric acid (Labscan, Thailand), polyvinylpyr-    „™â«‘∏’ MTT assay ‡æ◊ËÕ»÷°…“º≈¢Õß “√ °—¥®“°„∫
                                                                                                  ˆ
             rolidone 40,000 (Fluka, USA), BCA protein assay kit  ™–¡«ßμàÕ cell viability ¢Õ߇´≈≈å Caco-2

             (Pierce) (Thermo Fisher Scientific, USA), scintillation    Ù.Ò °“√‡æ“–‡≈’Ȭ߇´≈≈å Caco-2
             cocktail (Perkin Elmer, USA), HMG CoA reductase            ‡≈’Ȭ߇´≈≈å Caco-2 ¥â«¬Õ“À“√‡≈’Ȭ߇´≈≈å DMEM/
             assay kit (Sigma, USA), pancreatic lipase (Sigma, USA),  F12 ∑’Ë¡’ à«πº ¡¢Õß Ò% FBS ·≈– Ò% penicillin-strep-

             4- methylumbelliferyl oleate (Sigma, USA), pravastatin  tomycin ∫à¡„π incubator ∑’ËÕÿ≥À¿Ÿ¡‘ Û˜˚´ ·≈– CO  ı%
                                                                                                           2
             (Sigma, USA), orilstat (Roche, Korea)              ‡¡◊ËÕ‡´≈≈å Caco-2 ‡®√‘≠∑—Ë«®“π‡≈’È¬ß ∑”°“√ subculture ∑ÿ°
                                                                                                            2+
                 «— ¥ÿ«‘∑¬“»“ μ√å∑’Ë„™â ‰¥â·°à silica gel 60 (E. Merck,  Û-Ù «—π ‚¥¬ detach ‡´≈≈å¥â«¬ .Úı% trypsin „π Ca -,
                                                                   2+
             Garmany), Sephadex LH20 (GE Healthcare, Sweden)    Mg -free phosphate buffer ∑’Ë¡’ EDTA .Ú °./≈. ‡ªìπ
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