Tài liệu Nghiên cứu trích ly hoạt chất từ nấm linh chi và thử nghiệm hoạt tính sinh học của cao trích

ĈҤ,+Ӑ&48Ӕ&*,$73HCM 75ѬӠ1*ĈҤ,+Ӑ&%È&+.+2$ ----------------------- 1*Ð7+ӎ7+8ǣ/,1+ 1*+,Ç1&Ӭ875Ë&+/<+2Ҥ7&+Ҩ77Ӯ 1Ҩ0/,1+&+,9¬7+Ӱ1*+,ӊ0+2Ҥ77Ë1+ 6,1++Ӑ&&Ӫ$&$275Ë&+ (Extraction of biological activities from Ganoderma lucidum and examination of bioactivities) &KX\rQQJjQK.Ӻ THUҰ7+Ï$+ӐC 0mVӕ: 8520301 LUҰ19Ă17+Ҥ&6Ƭ 73+Ӗ&+Ë0,1+WKiQJ1 QăP &{QJWUuQKÿѭӧFKRjQWKjQKWҥL7UѭӡQJĈҥL+ӑF%iFK.KRD- Ĉ+4*73+&0 &iQEӝKѭӟQJGүQNKRDKӑF76+RjQJ0LQh Nam 3*6761JX\ӉQ+ӳX+LӃX &iQEӝFKҩPQKұQ[pW76+XǤQK1JӑF2DQK &iQEӝFKҩPQKұQ[pW761JX\ӉQ4XӕF7KҳQJ /XұQYăQWKҥFVƭÿѭӧFEҧRYӋWҥL7UѭӡQJĈҥL+ӑF%iFK.KRD- Ĉ+4*73+&0, WKiQJQăP 7KjQKSKҫQ+ӝLÿӗQJÿiQKJLiOXұQYăQWKҥFVƭJӗP 3*6761JX\ӉQ7XҩQ$QK- &KӫWӏFK 76+XǤQK1JӑF2DQK- Ӫ\YLrQSKҧQELӋQ 761JX\ӉQ4XӕF7KҳQJ- Ӫ\YLrQSKҧQELӋQ 769}1JX\ӉQĈҥL9LӋW- Ӫ\YLrQ 763KҥP7UӑQJ/LrP&KkX- 7KѭNê ;iFQKұQFӫD&KӫWӏFK+ӝLÿӗQJÿiQKJLi/9Yj7UѭӣQJ.KRDTXҧQOêFKX\rQ QJjQKVDXNKLOXұQYăQÿmÿѭӧFVӱDFKӳDQӃXFy &+Ӫ7ӎ&++Ӝ,ĈӖ1* +ӑWrQYjFKӳNê 75ѬӢ1*.+2$.Ӻ7+8Ұ7+Ï$+Ӑ& +ӑWrQYjFKӳNê i ĈҤ,+Ӑ&48Ӕ&*,$73+&0 75ѬӠ1*ĈҤ,+Ӑ&%È&+.+2$ &Ӝ1*+Ñ$;­+Ӝ,&+Ӫ1*+Ƭ$9,ӊ71$0 ĈӝFOұS- 7ӵGR- +ҥQKSK~F NHIӊM VӨ LUҰ19Ă17+Ҥ&6Ƭ +ӑWrQKӑFYLrQ1J{7Kӏ7KXǤ/LQK MSHV: 1970648 1Jj\WKiQJQăPVLQK 1ѫLVLQK73+ӗ&Kt0LQK &KX\rQQJjQK.ӻWKXұW+RiKӑF 0mVӕ I. 7Ç1 Ĉӄ 7¬, 7rQWLӃng ViӋt: 1JKLrQFӭXWUtFKO\KRҥt chҩt tӯ nҩPOLQKFKLYjWKӱ nghiӋm hoҥt WtQKVLQKKӑc cӫDFDRWUtFK 7rQWLӃng Anh: Extraction of biological activities from Ganoderma lucidum and examination of bioactivities. II. NHIӊM VӨ 9¬1ӜI DUNG: 7әQJTuan: 1ҩPOLQKFKLWULWHUSHQRLGKRҥWWtQKFӫDQҩPOLQKFKLSKѭѫQJSKiS HQ]\PH NӃW KӧS VLrX kP SKѭѫQJ SKiS ÿӏQK OѭӧQJ WULWHUSHQRLG SKѭѫQJ SKiS WKӱ QJKLӋPKRҥWWtQKVLQKKӑFFӫDFDRWUtFKYjTX\KRҥFKWKӵFQJKLӋP 7KӵFQJKLӋP - .KҧR ViW ҧQK KѭӣQJ FӫD WӯQJ \ӃX Wӕ WUtFK O\ ÿӃQ KjP OѭӧQJ WULWHUSHQRLG EҵQJ SKѭѫQJSKiSWUtFKO\enzyme NӃWKӧSVLrXkP - .KҧR ViW EӅ PһW QҩP WUѭӟF Yj VDX TXi WUuQK WUtFK O\ EҵQJ SKѭѫQJ SKiS WUtFK O\ enzyme NӃWKӧSVLrXkP - 7KtQJKLӋPVjQJOӑFFiF\ӃXWӕҧQKKѭӣQJWKHRP{KuQK3lackett±%XUPDQYjWӕL ѭXKyDÿLӅXNLӋQWUtFKO\WKHRP{KuQK%R[±Behnken; - 7KӱQJKLӋPKRҥWWtQKNKiQJXQJWKѭYjNKiQJNKXҭQ ,,,1*¬< GIAO NHIӊM VӨ: ,91*¬<+2¬17+¬1+1+,ӊM VӨ: 9&È1%Ӝ +ѬӞNG DҮN: 76+2¬1*0,1+1$0 PGS.TS NGUYӈN HӲU HIӂU TP. 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Sӕ liӋXÿѭӧc xӱ OêEҵng phҫn mӅm Design Expert ÿӇ WuPÿLӅu kiӋQWUtFKO\FKRKjPOѭӧng triterpenoid cao nhҩt. Ĉӗng thӡLFiFQKyP chӭFÿһFWUѭQJFӫDFDRWUtFKKuQKWKiLFҩXWU~FEӅ mһt nҩPYj WKjQKSKҫQFDRWUtFKÿѭӧc khҧRViWEҵng phә hӗng ngoҥi chuyӇQKyD)RXULHU)RXULHU ± transform infrared spectroscopy ± )7,5 NtQK KLӇQ YL ÿLӋn tӱ TXpW VFDQ HOHFWURQ microscopy ± 6(0YjVҳc NêNKtNKӕi phә (Gas chromatography ± mass spectrometry ± GC±MS). &DR WUtFK WҥL ÿLӅu kiӋn tӕL ѭX ÿѭӧc thӱ nghiӋm hoҥW WtQK VLQK KӑF WK{QJ TXD NKҧ QăQJNKiQJXQJWKѭWUrQEDGzQJWӃ EjRXQJWKѭӣ QJѭӡLOj+HS±G2, KB, MCF±Yj khҧ QăQJ NKiQJ NKXҭn cӫa triterSHQRLG ÿѭӧc thӱ nghiӋP WUrQ Escherichia coli, Staphylococcus aureus, Pseudomonas aeruginosa Yj Salmonella enterica bҵng SKѭѫQJSKiSÿRÿѭӡQJNtQKYzQJNKiQJNKXҭn. iv ABSTRACT In this thesis, triterpenoid was extracted from Vietnamese Ganoderma lucidum (G. lucidum) by the ultrasonic-assisted enzyme extraction (UAEE) method. The triterpenoid content was determined by ultraviolet±visible spectroscopy (UV±Vis). Chemical composition and heavy metal content in G. lucidum have been investigated, showing that G. lucidum has high fiber, carbohydrate and lipid content. In addition, the content of heavy metals such as As, Cd, Hg and Pb was within the allowable limit. The influence of each extraction condition such as enzyme concentration, etanol:material ratio, temperature, time and power on triterpenoid content was investigated by the method of alternating each variable to find a suitable range of variation. Then, conducting a simultaneous survey by experimental planning using the Plackett±Burman and Box±Behnken designs. The data were processed using Design Expert 11.0 software to find extraction conditions for the highest triterpenoid content. At the same time, the characteristic functional groups of the extract, the morphology of the surface structure of the fungus and the high composition of the extract were investigated by Fourier transform infrared spectroscopy (Fourier transform infrared spectroscopy ± FTIR), scanning electron microscopy (scan electron microscopy ± SEM), and gas chromatography ± mass spectrometry (GC±MS). The extract at optimal conditions was tested for biological activity through antitumor activity on three human cancer cell lines (Hep-G2 and MCF-7) and the antibacterial ability of triterpenoids was tested on Escherichia coli, Staphylococcus aureus, Pseudomonas aeruginosa, and Salmonella enterica by antibacterial ring diameter measurement. v LӠ,&$0Ĉ2$1 7iFJLҧ [LQFDPÿRDQOXұQYăQQj\OjF{QJWUuQKQJKLrQFӭu thӵc sӵ cӫDFiQKkQWiF giҧ Yjÿѭӧc thӵc hiӋQGѭӟi sӵ Kѭӟng dүn cӫa thҫ\76+RjQJ0LQK1DPYj3*676 NguyӉn Hӳu HiӃX3KzQJWKtQJKLӋm TrӑQJÿLӇPĈ+4*+&0&{QJQJKӋ +yDKӑc Yj'ҫXNKt.H\&(33/DE7UѭӡQJĈҥi hӑF%iFK.KRD- Ĉ+4*73+&0 &iF Vӕ liӋu, kӃt quҧ QJKLrQ Fӭu trong luұQ YăQ Qj\ Oj KRjQ WRjQ WUXQJ WKӵF FKѭD tӯQJÿѭӧFF{QJEӕ trong bҩt cӭ mӝWF{QJWUuQKQjR NKiFWUѭӟFÿk\0ӑi sӵ JL~S ÿӥ cho viӋFKRjQWKjQKOXұQYăQQj\ÿӅXÿmÿѭӧc cҧPѫQFiFWK{QJWLQWUtFKGүn trong luұQYăQQj\ÿӅXÿmÿѭӧc chӍ U}QJXӗn gӕc. 7iFJLҧ xin chӏXWUiFKQKLӋm vӅ QJKLrQFӭu cӫDPuQK 7iFJLҧ 1J{7Kӏ7KXǤ/LQK vi MӨC LӨC LӠI CҦM Ѫ1 .............................................................................................................. iii 7Ï07ҲT LUҰ19Ă1 ...............................................................................................iv ABSTRACT ...................................................................................................................v LӠ,&$0Ĉ2$1 .........................................................................................................vi MӨC LӨC ................................................................................................................... vii DANH MӨ&+Î1+ ......................................................................................................ix DANH MӨC BҦNG .....................................................................................................xi DANH MӨ&&È&7Ӯ VIӂT TҲT ........................................................................... xii ĈҺT VҨ1Ĉӄ.................................................................................................................1 &+ѬѪ1* TӘNG QUAN .........................................................................................2 1.1. Tәng quan vӅ nҩm linh chi ................................................................................2 1.1.1. Giͣi thi͏u chung ...........................................................................................2 7KjQKSK̯QKRiK͕c .....................................................................................3 1.2. HoҥWWtQKVLQKKӑc cӫa triterpenoid..................................................................8 .KiQJR[\KRi .............................................................................................. 8 .KiQJXQJWK˱ .............................................................................................. 9 .KiQJNKX̱n ............................................................................................... 10 &iFSKѭѫQJSKiSWUtFKO\KRҥt chҩt trong nҩm linh chi .............................. 11 3K˱˯QJSKiSWUtFK6R[KOHW .........................................................................11 3K˱˯QJSKiSYLVyQJ ..................................................................................12 3K˱˯QJSKiSWUtFKO\E̹QJO˱XFK̭WVLrXWͣi h̩n .......................................13 3K˱˯QJSKiSVLrXkP .................................................................................14 3K˱˯QJSKiSHQ]\PH ..................................................................................17 7uQKKuQKQJKLrQFӭXWURQJYjQJRjLQѭӟc ...................................................18 7URQJQ˱ͣc..................................................................................................18 1JRjLQ˱ͣc..................................................................................................18 1.5. MөF WLrX QӝL GXQJSKѭѫQJSKiS QJKLrQFӭX WtQK PӟL Yj ÿyQJ JyS Fӫa luұQYăQ ....................................................................................................................19 1.5.1. MͭFWLrX ......................................................................................................19 1.5.2. N͡i dung ......................................................................................................19 3K˱˯QJSKiSQJKLrQFͱu ............................................................................20 vii 1.5.4. ĈyQJJySFͯa lu̵QYăQ ...............................................................................28 &+ѬѪ1* THӴC NGHIӊM..................................................................................29 +yDFKҩt, dөng cө, thiӃt bӏ YjÿӏDÿLӇm thӵc hiӋn.........................................29 +RiFK̭t ......................................................................................................29 2.1.2. Thi͇t b͓ ........................................................................................................29 Ĉ͓DÿL͋m th͹c hi͏n lu̵QYăQ .......................................................................30 7KtQJKLӋm ........................................................................................................30 2.2.1. Kh̫RViWWKjQKSK̯QKRiK͕c trong ṋm linh chi ......................................30 2.2.2. Kh̫RViWV͹ ̫QKK˱ͧng cͯDFiF\͇u t͙ ÿ͇QKjPO˱ͫng triterpenoid..........33  3KkQ WtFK ÿ̿F WtQK KyD Oê WKjQK SK̯n cͯD FDR WUtFK Yj F̭X WU~F QJX\rQ li͏XWU˱ͣFYjVDXNKLWUtFKO\ .................................................................................42 2.2.4. Th͵ nghi͏m ho̩WWtQKVLQKK͕c cͯDFDRWUtFK .............................................43 &+ѬѪ1* KӂT QUҦ 9¬%¬1/8ҰN.................................................................45 7KjQKSKҫQKRiKӑc trong nҩm linh chi ........................................................45 3.2. ҦQKKѭӣng cӫDFiF\Ӄu tӕ ÿӃQKjPOѭӧng triterpenoid ............................... 46 3.2.1. ̪QKK˱ͧng cͯa tͳng y͇u t͙ .........................................................................46 3.2.2. ̪QKK˱ͧQJÿ͛ng thͥi cͯDFiF\͇u t͙ .......................................................... 52 3.2.3. T͙L˱XKRiÿL͉u ki͏QWUtFKO\WULWHUSHQRLG ...................................................55 ĈһFWtQKKRiOêFӫDFDRWUtFKYjFҩXWU~FQJX\rQOLӋu ..................................61 3KkQWtFKFiFQKyPFKͱFWURQJFDRWUtFK ...................................................61 7KjQKSK̯n ho̩t ch̭t trong cao WUtFK ........................................................62 3.3.3. C̭XWU~FE͉ m̿t cͯDQJX\rQOL͏XWU˱ͣFYjVDXNKLWUtFKO\ .........................63 +RҥWWtQKVLQKKӑF ............................................................................................ 63 .KiQJNKX̱Q ........................................................................................................63 .KiQJXQJWK˱ .....................................................................................................65 &+ѬѪ1* KӂT LUҰN ........................................................................................... 67 DANH MӨ&&È&&Ð1*75Î1+.+2$+ӐC.....................................................68 7¬,/,ӊU THAM KHҦO.........................................................................................102 PHӨ LӨC ...................................................................................................................112 viii DANH MӨ&+Î1+ +uQK1ҩm linh chi ................................................................................................... 3 +uQKCҩXWU~Fȕ-glucan SKkQOұp tӯ nҩm linh chi [13]............................................ 4 +uQK&{QJWKӭc cҩu tҥo cӫa squalen ....................................................................... 5 +uQK&iFWULWHUSHQRLGWKXӝFQKyPĮ-amyrin .......................................................... 6 +uQK&iFWULWerpenoid thuӝFQKyPȕ amyrin ........................................................... 7 +uQK&iFWULWHUSHQRLGWKXӝFQKyPlupeol ............................................................... 7 +uQK&ѫFKӃ NKiQJR[\KyDEҵQJFiFKFKRÿLӋn tӱ ................................................ 9 +uQKҦQKKѭӣng cӫDWULWHUSHQRLGÿӃQTXiWUuQKWӃ EjRFKӃWWKHRFKXNu .............. 10 +uQK&ѫFKӃ NKiQJNKXҭn cӫa triterpenoid ........................................................... 11 +uQKHӋ thӕQJWUtFK6R[KOHW ................................................................................ 11 +uQKThiӃt bӏ SKiPүXYLVyQJ ............................................................................ 12 +uQKGiҧQÿӗ pha cӫa CO2 .................................................................................. 14 +uQK ThiӃt bӏ VLrXkP .......................................................................................... 15 +uQKQXiWUuQKWUtFKO\KRҥt chҩWFyKӛ trӧ cӫDVLrXkP ..................................... 16 +uQK0{Wҧ hӋ thӕQJWUtFKO\EҵQJVyQJVLrXkPGҥng bӇ ................................... 16 +uQK4XiWUuQKSKiYӥ cҩXWU~FWӃ EjRFӫa enzyme ............................................ 17 +uQK0{KuQK%R[-Behnken ............................................................................... 22 +uQK6ѫÿӗQJX\rQOêPi\ÿRSKә89±Vis ....................................................... 23 +uQKCҩu tҥo thiӃt bӏ quang phә FTIR ................................................................ 24 +uQK6ѫÿӗ QJX\rQOêPi\VҳFNêJKpSNKӕi phә GC-MS ................................. 25 +uQK1JX\rQWҳc hoҥWÿӝQJPi\6(0 ................................................................ 26 +uQK1JX\rQWҳc thӱ nghiӋm hoҥWWtQKNKiQJXQJWKѭ ....................................... 26 +uQK4X\WUuQKWUtFK ly triterpenoid tӯ nҩm linh chi bҵQJSKѭѫQJSKiSenzyme kӃt hӧSVLrXkP .............................................................................................................. 34 +uQKPhә UV ± YLVYjÿѭӡng cong chuҭn cӫa axit ursolic ӣ ɉ݉ܽ‫ ݔ‬± 548 nm ..... 41 +uQKĈөc lӛ WUrQEӅ mһWÿƭDWKҥch ......................................................................... 44 +uQK+jPOѭӧng kim loҥi nһng trong nҩm linh chi* ............................................. 46 +uQKҦQKKѭӣQJFӫDQӗQJÿӝHQ]\PHÿӃQKjPOѭӧQJWULWHUSHQRLG ..................... 47 +uQKҦQKKѭӣng cӫa tӹ lӋ dung P{LQJX\rQOLӋXÿӃQKjPOѭӧng triterpenoid ... 48 +uQKҦQKKѭӣng cӫa nhiӋWÿӝ VLrXkPÿӃQKjPOѭӧng triterpenoid ..................... 49 +uQKҦQKKѭӣng cӫa thӡLJLDQVLrXkPÿӃQKjPOѭӧng triterpenoid .................... 50 ix +uQKҦQKKѭӣng cӫDF{QJVXҩWVLrXkPÿӃQKjPOѭӧng triterpenoid ................... 51 +uQK(a) BiӇXÿӗ SKkQSKӕi nӱa chuҭQYjE%LӇXÿӗ 3DUHWRFiFÿLӇm A, B, C, D, Yj( ................................................................................................................................ 55 +uQKҦQKKѭӣQJÿӗng thӡi cӫa nӗQJÿӝ enzyme YjWӹ lӋ HWDQROQJX\rQOLӋu ..... 58 +uQKҦQKKѭӣQJÿӗng thӡi cӫDF{QJVXҩWVLrXkPYjQӗQJÿӝ enzyme ............... 59 +uQKҦQKKѭӣQJÿӗng thӡi cӫa etanol: tӹ lӋ QJX\rQOLӋXYjF{QJVXҩWVLrXkP . 60 +uQK3. 11: Phә FTIR cӫDFDRWUtFK ............................................................................... 61 +uQK +uQKҧnh SEM cӫa quҧ thӇ nҩPOLQKFKLDEWUѭӟFYjFGVDXNKLWUtFKO\63 +uQK  : 9QJ ӭc chӃ cӫa (a) E. coli , (b) S. aureus , (c) P. aeruginosa Yj  G S. enterica .......................................................................................................................... 64 +uQKKhҧ QăQJӭc chӃ tӃ EjRXQJWKѭFӫa FDRWUtFK............................................ 65 x DANH MӨC BҦNG Bҧng 1. 1+jPOѭӧng chҩWNKRiQJYjYLWDPLQFyWURQJQҩm linh chi............................ 3 Bҧng 1. 2'zQJÿҫXWLrQFӫa ma trұn Hadamard khi N= 12, 20, 24 ............................ 21 Bҧng 2. 1: &iFKyDFKҩt sӱ dөng ...................................................................................29 Bҧng 2. 2: &iFGөng cө YjWKLӃt bӏ sӱ dөng ...................................................................29 BҧQJ&iF\Ӄu tӕ khҧRViW ......................................................................................35 Bҧng 2. 4: .KҧRViWҧQKKѭӣQJFӫDQӗQJÿӝHQ]\PH ...................................................35 Bҧng 2. 5: Khҧo ViWҧQKKѭӣng tӹ lӋ HWDQROQJX\rQOLӋu...............................................36 Bҧng 2. 6: KhҧRViWҧQKKѭӣng cӫa nhiӋWÿӝ .................................................................36 Bҧng 2. 7: KhҧRViWҧQKKѭӣng cӫa thӡi gian................................................................ 37 Bҧng 2. 8: KhҧRViWҧQKKѭӣng cӫDF{QJVXҩt............................................................... 37 Bҧng 2. 9: PhҥPYLFiFELӃQWURQJP{KuQK3ODFNHWW± Burman....................................38 Bҧng 2. 10: ThiӃt kӃ WKtQJKLӋPFKRP{KuQKVjQJOӑF3ODFNHWWí%XUPDQ ....................38 Bҧng 2. 11: Ma trұn yӃu tӕ PmKyDWKHRP{KuQK%R[±Behnken .................................39 Bҧng 3. 17KjQKSKҫQKRiKӑc trong nҩm linh chi* ....................................................45 Bҧng 3. 2: KӃt quҧ thӵc nghiӋPWKHRP{KuQK3ODFNHWW-Burman..................................52 Bҧng 3. 3: KӃt quҧ SKkQWtFKWKӕQJNr$129$ ........................................................... 53 Bҧng 3. 4: Sӵ biӃQÿӝng cӫDFiF\Ӄu tӕ khҧRViW .......................................................... 54 Bҧng 3. 5: KӃt quҧ thiӃt kӃ WKtQJKLӋm tӯ P{KuQK3ODFNHWW± Burman ........................55 Bҧng 3. 6: KӃt quҧ SKkQWtFK$129$ ..........................................................................56 xi DANH MӨ&&È&7Ӯ VIӂT TҲT 7ӯYLӃWWҳW AOAC DPPH FTIR GC±MS SEM ICP MS IC50 7LӃng Anh 7LӃQJ9LӋW Association of Official Agricultural HiӋp hӝLFiFQKjKyDKӑc Chemists SKkQWtFKFKtQKWKӭc 2,2-diphenyl-1-picrylhydrazyl Fourier ± transform infrared Phә hӗng ngoҥi chuyӇQKyD spectroscopy ± FTIR Fourier Gas chromatography ± mass SҳFNêNKt- khӕi phә spectrometry Scanning electron microscopy Inductively coupled plasma mass spectroscopy 4XpWNtQKKLӇQYLÿLӋn tӱ Quang phә nguӗn plasma cҧm ӭng cao tҫn kӃt nӕi khӕi phә Half Maximal Inhibitory Concentration NӗQJÿӝ ӭc chӃ 50% OD Optical density MұWÿӝ quang ROS Reactive Oxygen Species &iFJӕFR[\KRiWӵ do RSM Response surface methodology 3KѭѫQJSKiSEӅ mһWÿiSӭng UV-Vis Ultraviolet±visible spectroscopy Phә hҩp thө tӱ ngoҥi-khҧ kiӃn xii ĈҺT VҨ1Ĉӄ 1Jj\Qay, vӟi sӵ FăQJWKҷQJYjiSOӵc cӫD[mKӝi hiӋQÿҥLÿmҧQKKѭӣQJNK{QJQKӓ ÿӃn sӭc khoҿ cӫDFRQQJѭӡi. MһFGQKӳng tiӃn bӝ GRF{QJQJKӋ mang lҥLÿmOjPFKR cuӝc sӕng cӫDFRQQJѭӡi trӣ QrQGӉ GjQJKѫQQKѭQJFRQQJѭӡi vүQÿDQJWuPNLӃPFiF giҧi SKiSÿLӅu trӏ bӋnh thay thӃ NKiF FyQJXӗn gӕc tӯ tӵ QKLrQWӕW KѫQĈLӅX Qj\ÿm dүQÿӃn nhu cҫXQJj\FjQJWăQJÿӕi vӟLFiFWKҧRGѭӧFFyFKӭDFiFKRҥt chҩWFyKRҥt WtQK VLQK KӑF ÿѭӧc sӱ dөQJ ÿӇ QkQJ FDR Vӭc khӓH Yj Kӛ trӧ ÿLӅu trӏ bӋnh. Ӣ Trung Quӕc, nҩm liQKFKLÿmÿѭӧFQJKLrQFӭu rҩt nhiӅXYjVӱ dөQJQKѭPӝt loҥi thҧRGѭӧc ÿӇ trӏ bӋQKÿLӅXKzDKX\ӃWiSFKӕQJOmRKyDNpRGjLWXәi thӑWăQJWUtQKӟÿLӅXKzD hoҥWÿӝng tim mҥFKK{Kҩp, lӧi tiӇu, bә thұQYjWUӏ ÿDXQKӭF[ѭѫQJNKӟS&iFWKjQK phҫQ KyD Kӑc cy WURQJ Qҩm linh chi rҩW SKRQJ SK~ EDR JӗP FiF QKyP $[LW EpR VWHURLG DONDORLG SRO\VDFFKDULGH Yj WULWHUSHQRLG 7URQJ ÿy WKjQK SKҫQ Fy WiF Gөng Gѭӧc hӑF ÿһF WUѭQJ FKR Qҩm linh chi phҫn lӟn thuӝc vӅ QKyP WULWHUSHQRLG &iF QKj QJKLrQ FӭX ÿm QKұQ ÿӏnh rҵng WULWHUSHQRLG Fy FiF KRҥW WtQK VLQK KӑF QKѭ NKiQJ R[\ KyDEҧo vӋ gan, chӕng dӏ ӭng, chӕng khӕLXYjOjPJLҧPOѭӧQJFKROHVWHUROWURQJPiX &iF KRҥt chҩt trong nҩP OLQK FKL WKѭӡQJ ÿѭӧF WUtFK O\ EҵQJ SKѭѫQJ SKiS WUX\Ӆn thӕng sӱ dөQJGXQJP{LKӳXFѫKRһFQѭӟc QyQJQKѭQJQKӳQJSKѭѫQJSKiSWUrQFy nhiӅu hҥn chӃ QKѭKLӋu quҧ WUtFKO\WKҩSOjPELӃQWtQKFiFKRҥt chҩt, tӗn tҥLFiFGXQJ P{L KӳX Fѫ ÿӝc trong sҧn phҭP WKX ÿѭӧF 4Xi WUuQK WUtFK O\ EҵQJ SKѭѫQJ SKiS enzyme kӃt hӧp vӟL VyQJ VLrX kP Oj PӝW SKѭѫQJ iQ WKD\ thӃ tuyӋt vӡL FKR FiF TXi WUuQKWUtFKO\WUX\Ӆn thӕng. 3KѭѫQJSKiSWUtFKO\Eҵng SKѭѫQJSKiSHQ]\PHNӃt hӧp vӟLVyQJVLrXkP sӱ dөng GXQJP{LHWDQROFyPӝt sӕ WtQKFKҩWQKѭGXQJP{LVҥFKDQWRjQtWWӕQNpP YjWKLӃt bӏ WѭѫQJ ÿӕi thҩS QKѭQJ FKR WKӡi gian WUtFK O\ QJҳQ KjP OѭӧQJ WUtFK O\ FDR Vҧn phҭm VDXNKLWUtFKO\FyFKҩWOѭӧng tӕt, hҥn chӃ ÿѭӧc sӵ KuQKWKjQKFiFJӕc tӵ GRYjWKLӃt bӏ dӉ sӱ dөQJOjQKӳQJѭXÿLӇm nәi trӝLSKѭѫQJSKiSQj\'RÿyFyWKӇ ÿѭӧc lӵa chӑn FKRTXiWUuQKWUtFKO\YjSKkQWiFKFiFWKjQKSKҫn hoҥWWtQKWӯ FiFFk\WKXӕFTXê 9uYұy, luұQYăQQj\ÿѭӧc thӵc hiӋn vӟLWLrXÿӅ ³1JKLrQFӭXWUtFKO\KRҥt chҩt tӯ nҩPOLQKFKLYjWKӱ nghiӋm hoҥWWtQKVLQKKӑc cӫDFDRWUtFK´. 1 &+ѬѪ1* TӘNG QUAN 1.1. Tәng quan vӅ nҩm linh chi 1.1.1. Giͣi thi͏u chung Nҩm linh chi FyWrQNKRDKӑFOjGanoderma lucidum, thuӝc hӑ Ganodermataceae. NҩP OLQK FKL WKѭӡQJ Fy WrQ JӑL NKiF Oj Lingzi ӣ Trung Quӕc, reishi ӣ Nhұt BҧQ Yj yeongji ӣ +jQ4XӕFFzQFyӣ ViӋW1DPWKѭӡng gӑLOjWLrQWKҧo, nҩPWUѭӡng thӑSKkQ bӕ khҳSQѫLWUrQWKӃ giӟi, chӫ yӃXOjYQJQKLӋWÿӟLYjFұn nhiӋWÿӟLQKѭFKkXÈ NҩPOLQKFKLFySKkQORҥi khoa hӑFQKѭVDX[1]. *LӟL Fungi 1JjQK Basidiomycota /ӟS Agaricomycetes %ӝ Polyporales +ӑ Ganodermataceae Chi Ganoderma /RjL Ganoderma lucidum NҩPOLQKFKLWKѭӡng sӕng hoҥLVLQKWUrQQKLӅu loҥLFk\KRһFSKiWWULӇQWUrQFiFJLi thӇ OjJӛ mөc. Nҩm linh chi gӗm 2 phҫn cuӕng nҩPYjPNJQҩm. Cuӕng nҩPGjLKRһc ngҳQKuQKWUөÿѭӡQJNtQK± 3 cm. Cuӕng nҩPWKѭӡQJNK{QJSKkQQKiQKÿ{LNKL uӕn kh~FFRQJ/ӟp vӓ cuӕQJPjXÿӓ ± QkXÿӓ ± QkXÿHQEyQJNK{QJFyO{QJSKӫ suӕWWUrQPһWWiQQҩP0NJQҩm dҥng thұn ± gҫQWUzQÿ{LNKL[zHKuQKTXҥW7UrQPһt PNJFyYkQJҥFKÿӗQJWkPPjXVҳc tӯ YjQJFKDQK± YjQJQJKӋ ± YjQJQkX± YjQJFDP ±ÿӓ QkX± QkXWtP nhҹQEyQJOiQJQKѭYHFQL.KLJLjVүPPjXOӟp vӓ OiQJOӟp phҩn ÿӓ QkXWUrQEӅ mһWQJj\FjQJQKLӅX YjGj\ KѫQ.tFKWKѭӟF PNJQҩPWKѭӡng 5 ± 12 FPGj\± 3,3 cm. PhҫQÿӍnh cuӕQJWKѭӡng gӗ OrQKRһFKѫLO}P[2]. Nҩm linh chLOjORjLSKkQEӕ khҳSQѫLWUrQWKӃ giӟi. Ӣ ViӋt Nam, nҩPOLQKFKLFyWKӇ WuPWKҩy ӣ hҫu hӃWFiFWӍQKYQJQ~LWӯ /jR&DLÿӃQ/kPĈӗng. Nҩm linh chi chӫ yӃu sinh sҧn bҵQJEjRWӱ ӣ mһWGѭӟi cӫa quҧ thӇ. PhҫQFyFKӭFQăQJVLQKGѭӥQJFKtQKOj hӋ sӧi cӫa nҩm mӑc ҭn trong gӛ mөc hoһFÿҩt [3]. +uQKҧnh thӵc tӃ cӫa nҩm linh chi ÿѭӧc thӇ hiӋQQKѭKuQK 2 +uQK1: Nҩm linh chi Nҩm linh chi OjPӝt chҩWNtFKWKtFKKӋ thӕng miӉn dӏch mҥnh mӁ, bҧo vӋ hӋ thӕng miӉn dӏFKYjFѫWKӇ khӓLFiFFKҩWFyKҥi cho sӭc khӓHFRQQJѭӡi. NҩPOLQKFKLWKѭӡng ÿѭӧc sӱ dөQJWURQJÿLӅu trӏ FiFEӋQKQKѭYLrPJDQPmQWtQKYLrPWKұn, huyӃWiSFDR YLrPSKӃ quҧQYjFiFUӕi loҥn khӕi u [4, 5]. 1.1.2. ThjQKSK̯QKRiK͕c Nҩm linh chi ÿѭӧc sӱ dөQJ QKѭ Pӝt loҥi thuӕc truyӅn thӕng ӣ FiF QѭӟF SKѭѫQJ Ĉ{QJ GR Qy Fy FKӭD KѫQ  Kӧp chҩt hoҥW WtQK VLQK KӑF QKѭ WULWHUSHQRLG SRO\VDFFKDULGHQXFOHRWLGHVWHUROFiFD[LWEpRSURWHLQ[4]. 7ULWHUSHQRLGÿѭӧF[HPOj mӝt trong nhӳQJWKjQKSKҫn chӭa trong nҩm linh chi ÿѭӧFÿiQK JLiFDRYӅ hoҥW WtQK sinh hӑc, chiӃm mӝt sӕ OѭӧQJÿiQJNӇ trong nҩPYjWKӇ hiӋQFiFWiFGөng sinh hӑFQKѭ chӕQJXQJWKѭFKӕQJR[\KyDFKӕQJYLrPÿLӅXKzDPLӉn dӏch, chӕQJWăQJVLQKYjKҥ ÿѭӡng huyӃt [6, 7]. BҧQJ  WUuQK Ej\ KjP Oѭӧng chҩW NKRiQJ Yj YLWDPLQ Fy WURQJ nҩm. Bҧng 1. 1+jPOѭӧng chҩWNKRiQJYjYLWDPLQFyWURQJQҩm linh chi ChҩWNKRiQJYjYLWamin +jPOѭӧng (mg/100g) Canxi 832,00 Photpho 4,15 Sҳt 82,60 Magie 1,03 3 Natri 375,00 Kali 3,59 Vitamin B1 3,49 Vitamin B2 17,10 Vitamin B6 0,71 Niacin 61,90 Inositol 307,00 Choline 1,15 1.1.2.1. Polysaccharide &iF SRO\VDFFKDULGH Fӫa nҩP OLQK FKL Fy WiF dөng sinh hӑF QKѭ NKiQJ YLrP Kҥ ÿѭӡng huyӃt, chӕQJORpWFKӕng lҥi sӵ KuQKWKjQKNKӕLXYjWăQJFѭӡng khҧ QăQJPLӉn dӏch [8, 5]&yWUrQORҥLSRO\VDFFKDULGHÿѭӧFSKkQOұp tӯ nҩPOLQKFKLYjÿѭӧc cҩu tҥo bӣLFiFÿѫQYӏ JOXFRVHPDQQRVHJDODFWRVHIXFRVH[\ORVHYjDUDELQRVHWURQJÿy chӫ yӃu gӗm 2 loҥL FKtQK Oj JDODFWDQ Yj JOXFDQ Ĉһc biӋW JOXFDQ Fy OLrQ NӃW ȕ ȕJOXFDQOjKRҥt chҩt sinh hӑFFyQKLӅXF{QJGөQJTXêFyOӧLFKRFRQQJѭӡi [9, 10]ȕJOXFDQOjPӝt chuӛLFiFSKkQWӱ D-JOXFRVHOLrQNӃt bӣLOLrQNӃt (1,3)-ȕKD\-ȕ&iF sҧn phҭm tӵ QKLrQFyFKӭDȕ-JOXFDQÿmÿѭӧc sӱ dөng rҩWOkXQKѭQJȕ-glucan chӍ mӟi ÿѭӧFQJKLrQFӭu trong nhӳQJQăPJҫQÿk\ȕ-JOXFDQÿѭӧc sӱ dөnJÿӇ ÿLӅu trӏ mӝt sӕ bӋnh bao gӗP XQJ WKѭ EӋnh truyӅn nhiӉm, bӋnh tiӇX ÿѭӡQJ Yj FiF EӋnh tim mҥch [11, 12]. CҩXWU~FKyDKӑc cӫa hoҥt chҩWȕ-JOXFDQÿѭӧFWUuQKEj\QKѭӣ KuQK +uQK2: CҩXWU~Fȕ-glucan SKkQOұp tӯ nҩm linh chi [13] 4 1.1.2.2. Saponin 6DSRQLQFzQJӑLOjVDSRQRVLGOjPӝWQKyPJO\FRVLGOӟn, gһp rӝQJUmLWURQJWKӵc vұt. 6DSRQLQFNJQJFyWURQJPӝt sӕ ÿӝng vұWQKѭ+ҧLVkP&iVDR6DSRQLQFyPӝt sӕ WtQK chҩt ÿһc biӋWQKѭ/jPJLҧm sӭFFăQJEӅ mһt, tҥo bӑt nhiӅu khi lҳc vӟLQѭӟFFyWiF dөQJQKNJKRiYjWҭy sҥFK/jPYӥ hӗng cҫu ngay ӣ nhӳng nӗQJÿӝ rҩWORmQJ&yWKӇ tҥo phӭc vӟi cholesterol hoһc vӟLFiFFKҩt 3-b-K\GUR[\VWHURLGNKiF Saponin FzQJӑi OjVDSRQRVLG do chӳ /DWLQKVDSRFyQJKƭDOj[jSKzQJYuWҥo bӑWQKѭ[jSKzQJDӵa WUrQ FҩX WU~F KyD Kӑc cӫa phҫQ VDSRJHQLQ WKu VDSRQLQ ÿѭӧF FKLD WKjQK  QKyP Oӟn: VDSRQLQ WULWHUSHQRLG VDSRQLQ VWHURLG Yj VDSRQLQ VWHURLG DONDORLG &iF WULWHUSHQRLG trong nҩm linh chi thuӝFQKyPVDSRQLQWULWHUSHQRLG[14, 15]. 1.1.2.3. Triterpenoid 7ULWHUSHQRLG ÿѭӧF SKkQ Eӕ rӝQJ UmL WURQJ JLӟi thӵc vұW ÿӝng vұW Yj Wӗn tҥL Gѭӟi dҥng tӵ GR QKѭ FiF HVWHU KRһc dҥng glucosid. Khoҧng 140 loҥL WULWHUSHQRLG ÿm ÿѭӧc [iFÿӏnh trong nҩm linh chi, hҫu hӃt triterpenoid loҥLODQRVWDQH&KtQKFiFWULWHUSHQRLG Qj\OjPFKRQҩPOLQKFKLFyYӏ ÿҳQJ7ULWHUSHQRLGOjQKӳng hӧp chҩWÿѭӧc tәng hӧp tӯ ViX ÿѫQ Yӏ LVRSUHQ &iF WULWHUSHQRLG Fy Eӝ NKXQJ FKtQK Wӯ  QJX\rQ Wӱ carbon (C30H48WKѭӡng gһp trong thӵc vұW&iFWULWHUSHQRLGWӗn tҥLGѭӟi dҥng tӵ GRNK{QJFy phҫQÿѭӡQJFyFҩXWU~FYzQJPDQJPӝt sӕ QKyPFKӭFQKѭ±OH, ±O±& 2Ĉһc WtQKFKXQJOjWDQWӕWWURQJHWDQROPHWDQROtWWDQWURQJQѭӟc ngoҥi trӯ khi kӃt hӧp vӟi ÿѭӡQJÿӇ tҥRWKjQKJO\FRVLG>@ CҩXWU~FKRiKӑc cӫDFiFWULWHUSHQRLGWURQJQҩm linh chi dӵDWUrQODQRVWDQOjPӝt chҩt chuyӇQ KRi FӫD ODQRVWHURO &y WKӇ FKLD WULWHUSHQRLG WKjQK  QKyP GӵD YjR Fҩu WU~FNK{QJYzQJYzQJYjYzQJ1KLӅu triterpenoid chuyӇQKyDWӯ QKyPQj\VDQJ QKyPNKiF6ӵ chuyӇQKyDEDRJӗm viӋFWKD\ ÿәL QKyP±&22+WKjQKQKyP ±CH3 WѭѫQJӭQJWKjQKQKyP±CH2OH. x 7ULWHUSHQRLGNK{QJYzQJ 6TXDOHQÿѭӧFSKiWKLӋn lҫQÿҫu tӯ JDQFiPұS6TXDOXVQJRjLUDFzQÿѭӧFWuPWKҩy trong mӝt sӕ loҥi nҩm, dҫu olive [16]. +uQKWKӇ hiӋQF{QJWKӭc cҩu tҥo cӫa squalen. +uQK3: &{QJWKӭc cҩu tҥo cӫa squalen 5 x 7ULWHUSHQRLGYzQJ Thuӝc loҥi squalen oxit (mӑLYzQJ&ÿӅu ӣ cҩXKuQKWUDQVYӟi nhau): Lanosterol, SURWRVWHURODJQRVWHUROYjF\FORDXGLQRO 'ѭӟL WiF Gөng cӫD FiF HQ]\PH WKӵc vұt hoһc vi khuҭn, hoһF D[LW ORmQJ VDSRQLQ thӫ\ SKkQ WҥR WKjQK JHQLQ Yj SKҫQ ÿѭӡng. Genin do saponin thӫ\ SKkQ JӑL Oj sappogenin. &y WKӇ FKLD OjP  QKyP OӟQ Oj VDSRJHQLQ WULWHUSHQ Yj VDSRJHQLQ VWHURLG 9Ӆ phҫn ÿѭӡng, phә biӃQOj'±glucose, D±lactose, L±arabinose, L±ramnosa, axit galacturonic YjD[LW'±glucuronic. 1KyP VDSRJHQLQ WULWHUSHQRLG  YzQJ FKLӃm sӕ OѭӧQJ NK{QJ QKLӅu trong tӵ QKLrQ QKѭQJ ÿLӅX Oj QKӳng sapogenin quan trӑQJ ĈiQJ FK~ ê QKҩW Oj FiF FKҩt Panaxozid trong hӑ 1KkQ 6kP Panax gingseng &$ 0H\HU FiF &XUXUELWDFLQ WURQJ Kӑ bҫX Et (Cururbitaceae) [17]. 7ULWHUSHQRLGYzQJÿѭӧFFKLDOjPQKyPGӵDWUrQTXDQKӋ ÿmÿѭӧF[iFÿӏnh vӟi mӝt trong 3 chҩWWLrXFKXҭQOjĮ-myrin, ȕ-P\ULQYjOXSHRO[18]. 1KyP Į-amyrin: Į-DP\ULQ D[LW XUVROLF D[LW DVLDWLF D[LW [HQWRLF Yj D[LW TXLQRYLF +uQKWKӇ hiӋn cҩXWU~FFӫDFiFWULWHUSHQRLGQKyPĮ-amyrin. +uQK4: &iFWULWHUSHQRLGWKXӝFQKyPĮ-amyrin 6 1KyPȕ-amyrinȕ-myrin, axit oleanolic, axit albigenic, axit cincholic, erythrodiol, VR\DVDSRJHQRO$%Yj&1KyPQj\FKLӃPÿDVӕ trong sapogenin triterpen, phә biӃn nhҩW Oj VDSRQLQ Fӫa axit ROHDQROLF +uQK  WKӇ hiӋn cҩX WU~F FӫD FiF WULWHUSHQRLG QKyPȕ-amyrin [19]. +uQK5: &iFWULWHUSHQRLGWKXӝFQKyPȕ amyrin 1KyP/XSHRO/XSHROEHWXOLQWDUD[DVWHUROYjWKXEHURJHQLQ[20]+uQKWKӇ hiӋn cҩXWU~FFӫDFiFWULWHUSHQRLGQKyPOXSHRO +uQK6: &iFWULWHUSHQRLGWKXӝFQKyPlupeol Trong phҫQNK{QJSKkQFӵc cӫa quҧ thӇ nҩPOLQKFKLFyFiFWULWHUSHQRLGWӵ do vӟi KjPOѭӧng tӯ 3 ± 7KjQKSKҫQFKtQKOjFiFD[LWJDQRGHULF$%&')+,Yj- &KtQK FiF WULWHUSHQRLG Qj\ OjP FKR QҩP OLQK FKL Fy Yӏ ÿҳQJ NKiF YӟL FiF ORҥi nҩm NKiF+jPOѭӧQJWULWHUSHQRLGOjNKiFQKDXӣ FiFEӝ phұQNKiFQKDXYjJLDLÿRҥQSKiW triӇQNKiFQKDXFӫa nҩm [21]. 7URQJFiFWKjQKSKҫQKyDKӑFWUrQQKyPWULWHUSHQRLGOjPӝt trong sӕ nhӳQJGѭӧc chҩWTXê ÿѭӧFSKiWKLӋQÿҫXWLrQYӟi nhiӅXWiFGөng. Ӣ hҫu hӃWFiFQѭӟFSKiWWULӇn, WULWHUSHQRLG Oj FKҩW FKtQK ÿѭӧF GQJ OjP WLrX FKXҭQ ÿӇ ÿiQK JLi FKҩW Oѭӧng cӫD FiF ORjL QҩP OLQKFKL NKiF QKDX [22]. Triterpenoid chiӃP KjP Oѭӧng cao trong nҩm linh 7