Kev Tshawb Fawb Ntawm Cistanche Polysaccharides Tshem Tawm Dawb Radicals

Yog xav paub ntxiv:ali.ma@wecistanche.com

Wang Guowei Zhao Fang

(Lub Tsev Kawm Ntawv ntawm Chemistry thiab Chemical Engineering, Shihezi University, Beisi Road, Shihezi City, Xinjiang 832000)

Ntsiab lus: Cistanche; Cistanche polysaccharide,polysaccharidelos ntawm cistanche; dawb radical

Lub cev tsis tu ncua tsim ntau yam reactive oxygen radicals (ROS) nyob rau hauv cov txheej txheem ntawm oxidative metabolism ntawm lub neej kev ua ub no, uas yog intermediate khoom tsim los ntawm redox cov tshuaj tiv thaiv ntawm aerobic metabolism hauv tib neeg lub cev. Cov kev tshawb fawb tau pom tias dhau lawmdawb radicalsyuav tawm tsam lub neej macromolecular tshuaj thiab ntau yam organelles, ua rau muaj ntau yam kev puas tsuaj rau lub cev ntawm molecular, cellular, thiab cov ntaub so ntswg hauv nruab nrog cev, ua kom lub cev txoj kev laus thiab inducing ntshav txhaws, ntshav qab zib, thiab atherosclerosis. , qog nqaij hlav thiab lwm yam kab mob [1]. Raws li chemically synthesized antioxidants yog tshuaj lom rau tib neeg lub cev, kev tshawb nrhiav ntuj antioxidants nrog kev ua haujlwm siab thiab tsis muaj tshuaj lom uas tuaj yeem tshem tawm cov pa dawb radicals tau dhau los ua qhov kub ntawm cov tshuaj, zaub mov, thiab tshuaj engineering hauv xyoo tas los.

Cistanche [Cistanche deserticola (YC Ma.)]Feem ntau parasitizes ntawm cov hauv paus hniav ntawm Tamarisk, Nitraria, thiab Haloxylon ammodendron. Nws yog ib qho tshuaj suav tsis tshua muaj nyob hauv kuv lub tebchaws thiab hu ua"dej ginseng"[2]. Cistanche cistanche polysaccharide yog ib qho tseem ceeb ntawm kev ua haujlwm lom neeg ua haujlwm ntawmcistanche, tab sis nws tseem tsis tau qhia seb tuscistanche polysaccharide muaj cov nyhuv ntawm tshem tawm cov dawb radicals. Nyob rau hauv cov ntaub ntawv no, tus txheej txheem ntawm spectral tsom xam tau siv los txiav tximCov nyhuv scavenging ntawm Cistanche polysaccharide ntawm superoxide anion dawb radicals, hydroxyl dawb radicals, D PPH radicals, thiab singlet oxygen dawb radicals, zoo astxhawm rau ntsuas nws cov haujlwm antioxidant. Siv tag nrho cov peev txheej ntuj no los muab kev tshawb fawb hauv paus.

Nyem rau Cistanche qia rau dawb radical

Cov kev tshawb fawb noCistanche polysaccharidehauv vitroantioxidantkev ua si. Noj ascorbic acid raws li kev tswj hwm, Fenton zoo li cov tshuaj tiv thaiv tsim hydroxyl dawb radicals, autooxidation ntawm dopamine tsim superoxide anion.dawb radicals, cov tshuaj tiv thaiv ntawm sodium hypochlorite thiab hydrogen peroxide los tsim singlet oxygen, diphenyl iab hydrazine radicals raws li kev sim qauv, siv ultraviolet 2Visible spectrophotometry thiab chemiluminescence tsom xam txoj kev, thawj zaug los txiav txim qhov scavenging nyhuv.Cistanche polysaccharide ntawm superoxide anion dawb radicals, hydroxyl dawb radicals, D PPH ·dawb radicals(diphenyl hydrazino radicals) thiab singlet oxygen.Cov txiaj ntsig tau pom tias cov polysaccharides ntawm Cistanche muaj qhov cuam tshuam pom tseeb ntawm cov dawb radicals uas tau hais los saum toj no, thiab cov txiaj ntsig khawb tau zoo cuam tshuam nrog cov concentration ntawm polysaccharides.

Apparatus thiab reagents: UV 22401 ultraviolet-pom spectrophotometer (Shimadzu Corporation, Nyiv); Lum ntawm LB9507 ultra-sensitive raj luminometer (Berthold Technologies, Lub teb chaws Yelemees); R E252CS teb evaporator (Shanghai Yarong Biochemical Co., Ltd.) Instrument Factory); HW 2SY hluav taws xob qhov kub thiab txias dej da dej lauj kaub (Jintan Xinhang Instrument Factory); pH S23C acidity meter (Shanghai Precision Scientific Instrument Co., Ltd.); D ZF26020 lub tshuab nqus tsev vacuum (Shanghai Yiheng Technology Co., Ltd.).

Cistanche extractyog yuav los ntawm Kuitun Medicinal Materials Corporation. Luminol (Luminol, Sigma, USA), Dopamine (China National Institute for the Control of Pharmaceutical and Biological Products), FeSO 4, ED TA, Vitamin C, H 2O 2, NaClO, Na2CO 3, NaHCO 3, thiab lwm yam. domestic analytical reagent. Kev sim dej yog ion-exchanged distilled dej.

Covextraction txoj kevCistanche polysaccharides: 30g uacistanche hmoovthiab muab tso rau hauv 500mL Erlenmeyer lub hwj, thiab tsau rau hauv roj av ether (60-90 degree), ether thiab ib feem ntawm 80 feem pua ​​​​ethanol kub kua (30 degree) rau 6h, thiab cov residue. yuav evaporated rau dryness. . Cov dej lim dej tau muab tso rau hauv dej ntawm qhov sib piv ntawm 1: 10 rau 24 teev, thiab cua sov ntawm 30 degree kom tshem tawm 100 min. Tom qab kev tshem tawm tiav, lub supernatant tau txais los ntawm centrifugation. Lub supernatant tau concentrated nyob rau hauv lub siab txo thiab ces precipitated los ntawm kev ntxiv 3 zaug ntawm lub ntim ntawm 95 feem pua ​​ethanol, sawv rau 24 teev ntawm 4 degree, sau cov precipitate tom qab centrifugation, thiab nqus kom qhuav kom tau cov crude Cistanche cistanche polysaccharide. Qhov hnyav 1.00g ntawm crude polysaccharide, dissolve nws nyob rau hauv dej thiab dilute mus rau ib tug 50mL volumetric hwj, thiab teem cia rau kev ntsuam xyuas.

Kev xeem ntawm cistanche polysaccharide ntawm superoxide dawb radical (O) kev muaj peev xwm: kev siv dopamine auto-oxidation los tsim superoxide dawb radical kuaj lubcistanche polysaccharide ntawm superoxide dawb radical scavengingpeev xwm [3]. Ntxiv 0.1 hli olöL pH=10.12 mus rau lub kav dej qhuav thiab kawm tiav, 3.0mL ntawm carbonate buffer, 3.0mL dej, thiab 0.5mL ntawm cov qauv polysaccharide tov ntawm cov concentrations sib txawv, thiab muab tso rau hauv 30 degree dej da dej rau preheating rau 5 feeb. Ntxiv 1.0mL ntawm 2.0mm olöL dopamine preheated nyob rau hauv ib da dej ntawm 30 degree, sib tov tam sim ntawd, khaws cov tshuaj tiv thaiv ntawm 30 degree rau 5 feeb, thiab tam sim ntawd nres cov tshuaj tiv thaiv nrog 2 tee ntawm 10molöL HCl. Siv tsis raws li kev siv, ntsuas qhov nqus ntawm 221nm (Sample A), cov pab pawg dawb hloov pauv cov qauv nrog 0.5 mL dej (A dawb), thiab pawg tswj hwm hloov cov qauv nrog 0.5 mL ntawm ascorbic acid. Txhua tus qauv raug kuaj nyob rau hauv parallel 3 zaug thiab siv nws tus nqi nruab nrab. O superoxide radical scavenging rate (%)=(Ib qho piv txwv-A) öA blank × 100 feem pua ​​.

Kev sim ntawm Kev Txiav Txim Siab ntawm Cistanche Polysaccharide ntawm · OH Dawb Radical

EDTA 2Fe (Ê) 2 methylene xiav 2H 2O 2 system tau siv los ntsuas lub peev xwm ntawmCistanche polysaccharidetxhawm rau tshem tawm hydroxyl dawb radicals. Lub hauv paus ntsiab lus yog tias Feton zoo li cov tshuaj tiv thaiv ntawm ED TA 2Fe (Ê) thiab hydrogen peroxide tsim · OH radicals, uas txo cov yam ntxwv nqus siab ncov ntawm methylene xiav tshuaj ntawm 660nm. Nyob rau hauv ib koob ntawm 10mL xeem raj nrog stoppers, ntxiv 5 hli oLöL FeSO 4 tov 1.{{20}}mL, 5 hli oLöL EDTA tov 1.2mL, 0. {29}}3mol N aH 2 PO 4 2N a2H PO 4 (pH=7.4) buffer solution 2 .0mL, 20mm olöL hydrogen peroxide solution 1.3mL, Tom qab co, ntxiv 0.2mm olöL methylene xiav tshuaj 0.7mL, thiab tom qab ntawd ntxiv qhov sib txawv ntawm cov tshuaj ntsuam xyuas, vitamin C 1.0mL, thiab dilute nrog dej mus rau 10. 00mL, tom qab sib tov, incubate ntawm 37 degree rau 30min, ntsuas qhov absorbance ntawm txhua cov tshuaj ntawm 660nm, thiab hloov cov qauv kev daws teeb meem nrog dej hauv pawg dawb paug. Txhua tus qauv raug kuaj 3 zaug ua ke, thiab tus nqi nruab nrab raug coj mus kuaj. Tus qauv OH radical scavenging tus nqi (%)=[(A 2-A 1) ö(A 0-A 1) × 100 feem pua ​​. Hauv cov mis: A 0 - absorbance tus nqi ntawm lub raj tsis muaj kev puas tsuaj; A 1 - absorbance tus nqi ntawm lub raj puas; A 2 - tus nqi absorbance tom qab ntxiv cov qauv.

Kev sim ntawm lub peev xwm khawb ntawm Cistanche polysaccharide ntawm D PPH Dawb Radical

Noj {{{0}}}.1 mL ntawm Cistanche deserticola polysaccharide tov thiab ntxiv rau 5.0 mL ntawm D PPH · tov, sib tov sai, thiab tom qab sawv ntsug rau 10 feeb ntawm chav tsev kub, lub absorbance hloov ntawm 517 nm wavelength yog ntsuas [4]. Txhua tus qauv raug kuaj 3 zaug ua ke, thiab tus nqi nruab nrab raug coj mus kuaj. Tus qauv tshem tawm tus nqi ntawm D PPH · (%)=1- [(AB) öA 0] × 100 feem pua ​​.

Hauv cov qauv: A 0 - absorbance tus nqi ntawm D PPH yam tsis muaj qauv ntxiv (1.9mL D PPH ntxiv rau 0.1 mL 50 feem pua ​​​​ethanol); A - absorbance tus nqi tom qab cov tshuaj tiv thaiv ntawm cov qauv thiab D PPH; B - blank Tus nqi nqus ntawm cov qauv (piv txwv 0.1mL ntxiv rau 1. 9mL 50 feem pua ​​​​ethanol).

Cistanche polysaccharide scavenging singlet oxygen 1O 2 kuaj

Singlet oxygen (1O 2) yog tsim los ntawm cov tshuaj tiv thaiv ntawm NaClO thiab H 2O 2 ntawm 37 degree [5]. Cov tshuaj siv rau kev txiav txim siab ntawm 1O 2 yog 100L 2. 8mm olöL NaClO tov, 400L 5mm olöL NaOH tov, 300L 10mm olöL H 2O 2 daws, 50L 1mm olöL Luminol tov. Thaum lub sij hawm cov tshuaj tiv thaiv, cov tshuaj tiv thaiv yog ntxiv rau cov qauv kev daws teeb meem, thiab chemiluminescence analyzer cia li sau cov txheej txheem tshuaj tiv thaiv tib lub sijhawm. Noj NaClO 2H 2O 2 2N NaOH 2 Luminol raws li kev tswj tsis pub muaj. Pib cov tshuaj tiv thaiv, thiab ntxiv cov tshuaj tiv thaiv los ntawm lub reservoir ntawm lub chemiluminescence ntsuas rau cov qauv raj raws li tus nqi teev los ntawm qhov kev pab cuam. Tom qab ntawd, chemiluminescence siv ntawm cov tshuaj tiv thaiv sib tov tau sau rau ntawm daim ntawv kaw txhua txhua 5s, thiab txhua tus qauv raug ntsuas nyob rau hauv parallel 3 zaug, thiab qhov nruab nrab tus nqi raug coj mus. Tus nqi clearance yog xam raws li cov qauv hauv qab no: clearance rate (%)=(tswj luminous intensity-sample luminous intensity) ö tswj luminous siv × 100 feem pua ​​3.

Cov nyhuv scavenging ntawm cistanche polysaccharides ntawm superoxide dawb radicals

Cov tshuaj lom neeg ntawm superoxide dawb radicals yog active, uas yuav ua rau fatty acids ntawm biofilm los tsim peroxides, uas nyob rau hauv lem ua puas hlwb [6]. Hauv daim ntawv no, txoj kev dopamine tau siv los txiav txim siab lub peev xwm ntawm superoxide dawb radicals. Saib daim duab 1 rauLub peev xwm scavenging ntawm Cistanche ntawm superoxide dawb radicals.Cov txiaj ntsig tau pom tias:Cistanche polysaccharides tuaj yeem cuam tshuam cov superoxide dawb radicals tsimlos ntawm qhov pib-oxidation ntawm dopamine, thiab nyob rau hauv qhov ntsuas concentration ntau (0-8}m gömL), nws nce nrog qhov nce ntawm cov concentration, tab sis lub peev xwm khawb qis dua li ntawm ascorbic acid.

Tshem tawm cov nyhuv ntawm cistanche polysaccharide OH superoxide dawb radical

OH superoxide radical yog ib qho ntawm cov pa oxygen dawb radicals tshaj plaws hauv cov pa oxygen. Nws tuaj yeem cuam tshuam nrog yuav luag txhua yam macromolecule hauv cov hlwb uas muaj sia nyob, uas tuaj yeem ua rau lipid peroxidation ntawm lub cev nqaij daim tawv, protein polymerization, thiab depolymerization, nucleic acid fragmentation, thiab lwm yam. Cov txheej txheem biochemical ua rau cov ntaub so ntswg cell pathology, ua rau muaj ntau yam kab mob thiab. ua kom lub cev laus [7]. Daim duab 2 qhia txog lub peev xwm tshem tawm ntawm cistanche polysaccharides ntawm OH superoxide dawb radicals. Cov txiaj ntsig kev sim qhia tau tiascov polysaccharide ntawm Cistanche muaj qhov cuam tshuam zoo rau ·OHdawb radicals,thiab nws nce nrog qhov nce ntawm qhov concentration.

Lub peev xwm scavenging ntawm Cistanche cistanche polysaccharide ntawm D PPH dawb radical

D PPH radical (diphenyl-iab hydrazino radical) yog cov dawb radical ruaj khov, uas yog ntshav hauv cov kuab tshuaj organic thiab muaj zog nqus nyob ze 517nm. Tom qab ntxiv cov tshuaj antioxidants, ib feem ntawm cov dawb radicals yuav raug tshem tawm thiab kev nqus yuav tsis muaj zog, uas tuaj yeem siv los ntsuas cov tshuaj antioxidant ntawm cov tshuaj [8]. Lub peev xwm scavenging ntawm Cistanche polysaccharides ntawm D PPH dawb radicals yog pom nyob rau hauv daim duab 3. Cov kev sim tau pom tau hais tias lub peev xwm scavenging ntawm Cistanche deserticola polysaccharide ntawm D PPH dawb radicals yog zoo correlated nrog nws cov concentration, thiab lub peev xwm scavenging yog ntau dua li ntawm. ascorbic acid.

Lub peev xwm scavenging ntawm cistanche polysaccharides ntawm 1O 2 dawb radicals

Singlet oxygen zoo siab molecular oxygen nrog cov khoom siv tshuaj lom neeg. Txij li cov hluav taws xob tsis sib xws muaj qhov nyiam ua khub, cov dawb radicals qhia cov khoom siv tshuaj lom neeg ntau dua [9]. Daim duab 4 qhia txog kev tshem tawm cov peev txheej ntawm Cistanche deserticola polysaccharide ntawm 1O 2 dawb radicals. Cov txiaj ntsig kev sim qhia tau hais tias muaj peev xwm ntawmCistanche polysaccharideskom tshem tawm singlet oxygen qis dua li ntawm ascorbic acid nyob rau hauv thaj tsam ntawm {{0}} 0m gömL. Thaum cov concentration ntau dua 4.0m gömL, lub peev xwm tshem tawm Cistanche cistanche polysaccharides ntau dua li ntawm ascorbic acid.

Hauv kev xaus: Los ntawm kev sib txawv reactive oxygen-dawb radical systems, in vitro antioxidant kev ua ntawm Cistanche polysaccharide tau kawm kom ntxaws. Nws tuaj yeem pom los ntawm cov txiaj ntsig kev sim uas nyob rau hauv qhov kev ntsuam xyuas ntau yam, Cistanche polysaccharide extract muaj qhov cuam tshuam rau superoxide dawb radicals O, OH superoxide dawb radicals, D PPH dawb radicals, thiab 1 O 2 dawb radicals. Cov teebmeem tsis zoo ib yam. Cov txiaj ntsig tau pom tias Cistanche polysaccharide yog ib qho zoo heev exogenous natural antioxidant, uas tsim nyog rau kev tshawb fawb ntxiv, kev loj hlob, thiab kev siv dav.

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