392 CHINESE JOURNAL OF FOOD HYGIENE 2009 21 5 A(H1N1) 1 2 1 1 1 1 (1., 830026 ; 2., 830026) :A (H1N1) 1. MTT A (H1N1) 2. Read2muench A(H1N1) 3. (CPE) CPE MTT (TC 0 ) 20gΠml,TC 0 1 500gΠml,,(),,, A(H1N1), A(H1N1),, :;;;; ; A(H1N1) Research on Xinjiang SuoSuo Grape Extract Against Influenza A ( H1 N1) HE Hua, MA Long, XU Liang2jun, SHEN Jian2xin, YANG Hao2feng, LU Yao2qin (Urumqi Municipal Center for Disease Control and Prevention, Xinjiang Wulumuqi 830026, China) Abstract : Objective To research on Xinjiang SuoSuo grape extract against influenza A ( H1N1). MSeC MCSHEEHY ESI2 MSΠMS MSeC [7 ], MSeC, MSeC MSeC,MSeC,, LC2MSΠMS, [1 ] IP C,LISKDJ. Efficacy of cancer prevention by high2selenium garlic is primarily dependent on the action of selenium [ J ]. Carcinogenesis, 1995,16 :264922654. [2 ],,,. 4 MNNG [J ].,2008,37 (1) :90292. [3 ] GE HH, CAI XJ,TYSON JF, et al. Identification of selenium species in selenium2enriched garlic, onion and broccoli using high2performance ion chromatography with inductively coupled plasma mass spectrometry detection[j ]. Anal Commun,1996,33 :2792281. [4 ],,. [J ].,2005,34 (3) :3712374. [5 ] UDEN PC, BOAKYE HT, KAHAKACHCHI C, Tyson JF. Selective Detection and Identification of Se Containing Compounds review and recent Developments[J ]. J Chromatogr A, 2004,1050 (1) :85293. [6 ],,. [J ].,2007,35 (2) :2442246. Method 1. Use MDCK cell culture and MTT method on Xinjiang SuoSuo grape polysaccharide, SuoSuo grape flavonoids, total terpene SuoSuo grapes [7 ] MCSHEEHY S, YANG WENJ IE, PANNIER F, et al. Speciation analysis of selenium in garlic by two2dimensional high2performance liquid chromatography with parallel inductively coupled plasma mass spectrometric and electrospray tandem mass spectrometric detection [J ]. Analytica Chimica Acta, 2000, 421 :1472153. [:2008-12 - 26 ] :R15 ;TS20715 ;TS20713 ;Q503 :A :1004-8456 (2009) 05-0389 - 04 :
A(H1N1) 393 against influenza virus A (H1N1) 2type role. 2. Use read2muench method to calculate the TICD 50 in influenza virus A (H1N1) 2 type MDCK cells. 3. By observing cytopathic effect ( CPE), the calculation CPE cytopathic effect inhibition rate, MTT determination of drugs on cells, such as the protection rate to evaluate the SuoSuo grape polysaccharide, total flavonoids, total triterpenoid the effect of anti2influenza virus. Results Determine the total flavonoids of grape Suo Suo largest concentration of non2toxic (TC 0 ) to 20gΠml, Suo Suo total triterpenoid grape TC 0 for the 1 500gΠml. Flavonoids, polysaccharides extract directly inactivated with relatively good, but this triterpenoid, and the role of ribavirin was not obvious. Flavonoids, polysaccharides, triterpenes have therapeutic effects, and the triterpenoid and flavonoidsπs effect of therapeutic than the polysaccharide, the therapeutic effect of ribavirin than flavonoids, polysaccharides, triterpene. Conclusion After studying the characteristics of plant resources in Xinjiang Suo Suo grape extract of anti2influenza virus A ( H1N1) subtype of the role is a multi2 passing, In vitro studies have shown by the different active ingredients of total flavonoids of suo suo grape, suo suo grape polyphenols, Suo Suo grape triterpenoid have a better treatment results to prevention of the influenza virus A ( H1N1) subtype, directly inactivated treatment, can be regarded as anti2influenza medicine resources for further development and utilization. Key word : Flavones ;Polysaccharides ;Triterpenes ;Vitis ;Plant Extracts ;Anti2influenza Virus( H1N1),,,, [1 ], (5 %10 %) (40 %60 %) [2 ],() [3 ],,,,(oleanolic acid, OA) [4,5 ] ; OA [6 ] (proanthyocyanidin, OPC) (Vitis vinifera L. ) SUOSUO 8119 % ;,, (vitaceae),,,,,,,,,,,, [7 ],, ;,, ;, [8 ], [9 ],OA [10,11 ], [4,12-14,16 ], 1 1. 1 1. 1. 1 (MDCK), 1. 1. 2 AΠΠ139Π2005 H1N1,, 1. 1. 3,, 070106, 1. 1. 4,, -, 4812 % ;, 2110 % 11115 019 %( ) L - ( GIBCO) DMEM ( GIBCO) Trypsin ( GIBCO) (BBI) HEPES (Sigma) ( ) A ( H1N1) RDE( ) 112 11211 10-1 10-2 10-3 10-4 10-5 10-6 10-7 10-8 10-9
394 CHINESE JOURNAL OF FOOD HYGIENE 2009 21 5 10-10 10, 96, (CPE) (TCID 50 ), (TCID 0 ) 11212 MDCK : 2 500 500 100 20gΠml, 2 500 500 100 20gΠml, 7 500 1 500 300 60gΠml MDCK 96,,100 lπ, 4, 37 5 %,,35 d (TC 0 ) : 37 500 7 500 1 500300gΠml 4, 11213 2, 3, (1 ) (2 ) (3 ) :1 :, 100 lπ, 24 h,, 100 TCID 50,100 lπ,2 h,,; 2 :2 h, 2 h,,;3 : 2 h,, 4, 37 5 %,,35 d CPE + + + + + + +, CPE, MTT 11214 1121411, - (0 %) + (125 %) + + (25 %50 %) + + + (50 %75 %) + + + + (75 %100 %), 0 1 2 3 4,, CPE [16 ] :CPE = (CPE - CPE )Π CPE 1121412 :570 nm MTT, MDCK [16 ] := ( A - A )Π A 2 211 TICD 50 READ2MUENCH [1 ] A (H1N1) MDCK TICD 50 10 4 Π011 ml, TICD 0 10 9 Π011 ml, 100 TICD 50 Π011 ml 212 MDCK,,(TC 0 ) 20gΠml, 3 d,; 100gΠml, 20gΠml, 20 1 500gΠml, 300gΠml,1 1 MDCK(CPE) ( gπml) (h) 24 48 72 2500 500 100 20 - - - 2500 500 100 20 - - - 7500 1500 - - - 300 - - - 60 - - - 7500 1500 300 - - - - - - :- MDCK MDCK 213 A(H1N1) 21311 1, CPE 100100 %, CPE,12150 % ;2,CPE 80100 %, CPE ;3 CPE,, 93175 % 1 21312 A ( H1N1) 1 2, CPE 100100 %, CPE
A(H1N1) 395 21313 MDCK CPE, MTT A= 570 nm, 1 MDCK, ;2 MDCK,MDCK ;3 MDCK,,3 21314 A ( H1N1) 1 : 2,20gΠml 10gΠml 115, 2 :10gΠml, ;3 :20 10 750 300gΠml 115,,4 1 A(H1N1) MDCK (CPE) ( 400) ;3 CPE, CPE 2 3 311,,,, (VTT) (OA), [17 ] OA S2180 [18 ],,,,,,,,,,,,,, [19 ] RNA ;, [20 ] [21 ],, (polysaccharides),,,,,, [22 ]
396 CHINESE JOURNAL OF FOOD HYGIENE 2009 21 5 ( gπml) 1 CPE 2 A(H1N1) CPE ( %) 2 CPE CPE ( %) 3 CPE CPE ( %) 20 1 0 0 1 75100 1 2 0 0 81125 0 1 0 1 87150 10 1 0 0 0 93175 1 0 1 0 87150 1 0 1 2 75100 5 1 1 0 0 87150 1 1 1 1 75100 1 1 2 2 62150 20 1 1 1 0 81175 1 2 1 1 68175 2 1 1 2 56125 10 0 0 0 0 100100 0 1 1 0 87150 0 1 0 2 68175 5 0 0 1 0 93175 0 2 2 0 75100 1 0 1 1 81125 1500 4 4 4 3 6125 3 4 4 4 6125 2 1 0 1 75100 750 4 3 4 3 12150 3 3 4 4 12150 0 0 1 0 81125 375 3 4 4 4 6125 4 3 3 4 18175 1 2 3 1 56125 4 3 2 4 18175 4 3 3 4 12150 0 1 0 0 93175 0 0 0 0 0 0 0 0 0 0 0 0 4 4 4 4 4 4 4 4 4 4 4 4 3 MDCK ( gπml) 1 ( %) 2 ( %) 3 ( %) 20 7215 8014 8515 10 8916 8716 7815 5 7818 8313 6317 20 7913 5914 6814 10 9313 8512 8316 5 8719 6311 7012 1500 110 114 38216 750 213 217 9119 375 914 618 1410 300 212 715 34818 750 215 312 10111 375 916 711 2412 300 215 712 33814 GARIE [23 ], [24 ],,,,, [25 ],, 4 A(H1N1) (HA) ( gπml) 1 log 2 log 3 log 20 01226 11881 01526 11581 01526 11581 10 01075 21032 01451 11656 01828 11279 5 01150 11957 01451 11581 11054 11504 20 01150 11957 01903 11204 11204 01903 10 01000 21107 01602 11505 01527 11580 5 01075 21032 01752 11355 11054 11053 1500 21107 01000 21107 01000 01753 11354 750 21107 01000 21107 01000 01075 21032 375 21107 01000 21107 01000 11500 01607 300 11806 01301 11806 01301 01000 21107 21107 21107 21107 - - - :- HA,, 312 MDCK,,3,,,,CPE 80 %, ;,
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