Party1: Anti-inflammatory Thiab Antioxidant Properties Of Finger Millet (Eleusine Coracana (L.) Gaertn.) Ntau yam Cultivated hauv Sri Lanka

Yog xav paub ntxiv. tiv taujtina.xiang@wecistanche.com

Kev nthuav dav ntawm cov kab mob inflammatory-mediated thiab oxidative stress-associated kab mob yog nce thoob ntiaj teb, tsim kom muaj kev xav tau ntxiv rau qhov tshiab ntawmanti-inflammatorytus neeg sawv cev thiabantioxidants. Txoj kev tshawb no tau tsom mus rau kev txiav txim siab hauv vitro arachidonate {{0}} lipoxygenase (A5-LOX), xanthine oxidase (XO), hyaluronidase, oxidative burst inhibitory kev ua ub no, thiab antioxidant zog ntawm Ravi, Rawana , thiab Oshadha ntiv tes millet ntau yam siv ethanolic thiab methanolic rho tawm. Ntawm tag nrho cov extracts, cov methanolic extract ntawm Oshadha nthuav tawm qhov siab tshaj plaws A5-LOX (ICsvalue∶ 484.42ug/ml) thiab XO (ICsvalue∶764.34ug/ml) inhibitory kev ua ub no. Tag nrho cov extracts pom tsawg dua 50 feem pua ​​​​hyaluronidase inhibitory kev ua haujlwm ntawm 1 mg / ml concentration, Methanolic extracts pom muaj peev xwm inhibitory muaj peev xwm ntawm cov pa oxygen reactive (ROS) tsim los ntawm tag nrho cov ntshav phagocytes, nrog ICs0cov nqi sib xws nruab nrab ntawm 26.9 thiab 27.7 ug / ml, piv rau ibuprofen (IC.value: 11.18ug / ml). Tag nrho cov extracts pom muaj zog inhibition ntawm ROS ua los ntawm polymorphonuclear neutrophils cais los ntawm tib neeg cov ntshav thaum piv rau ibuprofen (ICsvalue: 2.47 ug / ml) thiab ICsvalues ​​ntawm methanolic thiab ethanolic extracts ranged ntawm 0.29 mus rau 0.47 ug / ml thiab 1.70 ug / ml thiab 1.35 ug / ml. raws. Tag nrho cov extracts muaj ntau npaum li cas ntawm phenolic tebchaw nrog rauflavonoidsthiab muaj peev xwm mus khawb 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic) acid (ABTS)cation,2,2-diphenyl-1-picryl-hydrazyl( DPPH), thiab oxygen radicals. Tsis tas li ntawd, lawv muaj peev xwm txo tau cov hlau ions thiab chelate hlau ions terminating radical generating tshua. Qhov no yog thawj tsab ntawv ceeb toom ntawm A5-LOX, XO, hyaluronidase, thiab oxidative tawg inhibitory zog ntawm ib qho kev rho tawm ntawm txhua tus ntiv tes millet ntau yam cog hauv Sri Lanka. Cov kev tshawb pom tau qhia txog lub peev xwm ntawm kev siv cov ntiv tes millet rho tawm los ua qhov chaw ntawm cov tshuaj tiv thaiv kab mob. Tsis tas li ntawd, cov kev tshawb pom tau qhia tias Ravi, Rawana, thiab Oshadha ntau yam yog qhov zoo ntawm antioxidants. Yog li ntawd, kev noj cov ntiv tes millet ntau yam tsis tu ncua tuaj yeem ua lub luag haujlwm tseem ceeb hauv kev tiv thaiv thiab kev noj haus ntawm cov kab mob oxidative kev nyuaj siab.

Nyem qhov no kom paub ntau yam khoom

1. Taw qhia

Kev mobyog kev tiv thaiv tiv thaiv ntawm covtiv thaiv kab moblub kaw lus kom tshem tawm cov teeb meem stimuli xws li cov kab mob, cov hlwb puas, thiab kev tsis haum tshuaj lossis tshuaj lom neeg nrog rau kev pib kho. Txawm hais tias o yog ib qho lus teb yog tias tsis tswj tau, kev mob ntau dhau ua rau muaj ntau yam mob hnyav thiab mob ntev. Tam sim no, feem ntau ntawm cov kab mob inflammatory-mediated yog nce thoob ntiaj teb, tsim kom muaj kev xav tau tshiab thiab muaj txiaj ntsig zoo los tiv thaiv kab mob [1, 2].

Arachidonate 5-lipoxygenase(A5-LOX) yog ib qho enzyme tseem ceeb uas koom nrog hauv biosynthesis ntawm cov neeg muaj peev xwm kho mob ntawm cov kab mob inflammatory thiab kev tsis haum tshuaj. Nws catalyzes tsim cov leukotrienes los ntawm arachidonic acid [3]. Leukotrienes yog endogenous mediators nyob rau hauv lub pathogenesis ntawm ib tug xov tooj ntawm inflammatory kab mob xws li mob hawb pob, ua xua rhinitis, mob bronchitis, rheumatoid mob caj dab, inflammatory plob tsis so tswj kab mob, thiab ua xua [3,4]. Yog li ntawd, inhibition ntawm A5-LOX yog qhov tseem ceeb hauv kev tiv thaiv thiab kev kho mob ntawm ntau yam kab mob.

Xanthine oxidase (XO) catalyzes oxidation ntawm hypo-xanthine rau xanthine thiab xanthine rau uric acid[5]. XO inhibitors thaiv cov biosynthesis ntawm uric acid thiab thiaj li txo cov qib ntawm uric acid thiab vascular oxidative kev nyuaj siab. Thaum lub sij hawm catalytic ua ntawm XO, reactive oxygen hom (ROS) yog tsim, thiab yog li ntawd, XO plays lub luag hauj lwm pathogenetic nyob rau hauv lwm yam inflammatory kab mob thiab. Yog li ntawd, XO inhibitors yog qhov tseem ceeb hauv kev kho mob ntawm ntau yam kab mob inflammatory nrog los ntawm kev ua haujlwm catalytic ntawm XO [2,5].

Hyaluronidase yog ib qho lysosomal enzyme uas catalyzes hydrolysis ntawm mucopolysaccharides xws li hyaluronic acid. Hauv rheumatoid mob caj dab, hyaluronidase ntau dhau degrades hyaluronic acid thiab txo nws cov nqi thiab molecular hnyav, ua rau mob caj dab [6]. Inhibition ntawm hyaluronic acid degradation yog qhov tseem ceeb thiab tseem ceeb hauv kev tswj hwm hyaluronidase-mediated pathological condi-tions [7].

Nyob rau hauv cov mob inflammatory, lub cev tiv thaiv kab mob yog attracted rau qhov chaw ntawm o. Thaum muaj kev tiv thaiv thiab kev tiv thaiv kab mob, NADPH oxidases nyob hauv lub cev tiv thaiv kab mob tau qhib thiab tsim ROS hauv ntau qhov tsim kom muaj oxidative tawg [1, 8]. Tsawg thiab nruab nrab ntawm ROS muaj txiaj ntsig zoo hauv cov txheej txheem physiological sib txawv. Txawm li cas los xij, overproduction ntawm ROS deregulates cellular zog uas nyob rau hauv lem txhim kho tus mob inflammatory [8,9]. Yog li, inhibition ntawm ROS-induced oxidative tawg yog ib qho kev kho mob los tiv thaiv thiab tswj cov kab mob inflammatory-mediated.

Thaum oxidative phosphorylation,dawb radicalssuav nrog ROS yog tsim los ntawm cov khoom lag luam [10] .Ntxiv rau cov cellular metabolism hauv ib txwm muaj, muaj ntau lwm yam uas ua rau kev tsim cov dawb radicals, thiab yog tias tus nqi tsim tawm ntawm cov dawb radicals ntau tshaj tus nqi ntawm neutralization, oxidative kev nyuaj siab. yog tsim, thiab nws tseem ceeb contributes rau tag nrho cov kab mob inflammatory. Txij li cov tshuaj antioxidants muaj peev xwm tiv thaiv kev tsim cov dawb radicals thiab quenching dawb radicals, qhov sib npaug ntawm antioxidants thiab dawb radicals yog qhov tseem ceeb los tswj lub cev kev ua haujlwm kom zoo [8,11,12].

Finger millet(Eleusine coracana(L.)Gaertn.) yog qhov tseem ceeb tshaj plaws me me nplej nyob rau hauv lub tropics, thiab ntau yam kev kho mob ntawm cov ntiv tes millet tau tshaj tawm yav dhau los [13-17]. Txawm li cas los xij, rau cov ntiv tes millet ntau yam uas tam sim no cog thiab noj hauv Sri Lanka, tsis muaj pov thawj tshawb fawb txog kev kho mob.

thiab muaj peev xwm muaj txiaj ntsig kev noj qab haus huv rau cov neeg siv khoom. Yog li ntawd, nws yog ib qho tsim nyog los kawm txog cov tshuaj tiv thaiv kab mob thiab tshuaj tua kab mob ntawm Sri Lankan ntiv tes millet ntau yam thiab kom paub txog lawv lub peev xwm los txhim kho cov khoom noj khoom haus thiab kev noj qab haus huv ntawm cov neeg siv khoom. Qhov tshwm sim ntawm cov kab mob inflammatory-mediated thiab oxidative kev ntxhov siab cuam tshuam nrog cov kab mob yog nce thoob ntiaj teb, tsim kom muaj kev xav tau tshiab rau cov khoom siv tshuaj tiv thaiv kab mob thiab cov tshuaj antioxidants[1, 2]. Nrog rau qhov keeb kwm yav dhau los no, txoj kev tshawb fawb tam sim no tau tsom mus rau kev ntsuam xyuas Sri Lankan ntiv tes millet ntau yam rau hauv vitro los tiv thaiv cov khoom hauv cov ntsiab lus ntawm A5-LOX, XO, hyaluronidase, oxidative burst inhibitory kev ua ub no, thiab cov khoom siv tshuaj tua kab mob antioxidant siv cov tshuaj antioxidant sib txawv. .

2. Cov ntaub ntawv thiab cov txheej txheem

2.1.Sample Sau thiab Npaj. Ntiv tes millet ntau yam uas tau pom zoo rau kev cog qoob loo hauv Sri Lanka los ntawm Lub Tsev Haujlwm Saib Xyuas Kev Ua Liaj Ua Teb, uas yog Ravi, Rawana, thiab Oshadha, tau sau los ntawm Field Crops Research and Development Institute (FCRDI), Mahailuppallama, Sri Lanka. Cov ntau yam no tau cog rau hauv qhov chaw sim nyob rau hauv Low Country Dry Zone, ntawm FCRDI, Mahailuppallama, thiab cov noob tau ntawv pov thawj los ntawm Seed Certification Service ntawm Department of Agriculture, Sri Lanka.

Ntiv tes millet noob tau dehulled (TM 05C, Satake Cor-poration, Nyiv), thiab hmoov nplej los ntawm tag nrho cov nplej tau los ntawm milling (Pulverisette 14, Fritsch, Lub teb chaws Yelemees) thiab dhau los ntawm 0.5 hli sieve. Cov hmoov nplej tag nrho tau muab rho tawm nrog ethanol thiab methanol cais. Tag nrho cov hmoov nplej (100g) tau muab rho tawm nrog cov kuab tshuaj (400 ml) hmo ntuj ntawm chav tsev kub (28 ± 2 degree) siv lub tshuab sib nqus thiab centrifuged ntawm 4000 rpm rau 20min. Lub supernatant tau sau sib cais, thiab cov seem tau muab rov ua dua ob zaug siv tib lub sijhawm. Cov supernatants tau muab tso ua ke thiab evaporated kom qhuav nyob rau hauv qhov txo qis ntawm 40 degree siv lub evaporator teb (R-114, Büchi Labortechnik AG, Switzerland). Cov kuab tshuaj tsis muaj kuab tshuaj tau muab khaws cia rau hauv cov thawv ntim airtight ntawm -20 degree kom txog thaum siv rau kev tshuaj xyuas.

2.2.Enzymes, Tshuaj, thiab khoom siv. Folin-Ciocalteu phenol reagent, aluminium chloride, 2,4, 6-Tri(2-}pyridyl)-s-triazine (TPTZ), 2,2'-azobis (2-}amidinopropane) dihydrochloride (AAPH), 2, 2-diphenyl-1-picrylhydrazyl (DPPH), quercetin, 6-hydroxy-2,5,7,8-tetramethylchroman{19} }carboxylic acid (Trolox), ethylenediaminetetraacetic acid (EDTA), 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) diammonium ntsev (ABTS), fluorescein, 3-( 2-Pyridyl)-5,6-diphenyl-1,2,4-triazine-p, pdisulfonic acid monosodium ntsev

hydrate (ferrozine), arachidonate 5-lipoxygenase, linoleic acid, baicalein, xanthine oxidase, xanthine, allopurinol, hyaluronidase, hyaluronic acid sodium ntsev, p-dimethylaminobenzaldehyde, gallic acid, tannic sulfenoxide, thiab diupdyl sulfen. Sigma-Aldrich, MO, USA. HBSs* plus thiab HBSS tau yuav los ntawm Thermo Fisher Sci-entific Inc., Massachusetts, USA. Zymosan tau yuav los ntawm Wako Pure Chemical Industries Ltd., Osaka, Nyiv.

Luminol (3-aminophthalhydrazide) tau yuav los ntawm Alfa Aesar GmbH & Co KG, Karlsruhe, Lub teb chaws Yelemees. Lymphocyte cais nruab nrab tau yuav los ntawm MP Biomedicals, Inc., Ohio, USA. Tag nrho lwm yam tshuaj thiab reagents siv nyob rau hauv qhov kev sim yog ntawm ACS, HPLC, thiab analytical qib. Txhawm rau txiav txim siab cov pa oxygen radical absorbance muaj peev xwm, fluorescence microplate nyeem ntawv (SpectraMax-Gemini EM, Molec-ular Devices Inc., USA) tau siv. Rau kev soj ntsuam chemiluminescence, luminometer (Luminoskan RS, Labsystems, Finland) tau siv. Rau tag nrho lwm yam bioassays, tus nyeem ntawv microplate (Spectra-Max Plus384, Molecular Devices Inc, USA) tau siv.

2.3.Kev txiav txim siab tag nrho cov ntsiab lus Phenolic. Tag nrho cov ntsiab lus phenolic (TPC) tau txiav txim siab raws li txoj kev Folin-Ciocalteu 【18】. Ntiv tes millet extract (20ul) yog tov nrog 110ul ntawm freshly npaj 10 zaug diluted Folin-Ciocalteu reagent thiab 70ul ntawm sodium carbonate tov (10 feem pua ​​w/v) thiab incubated ntawm chav tsev kub (RT) rau 30min. Absorbance tau ntsuas ntawm 765 nm. Gallic acid tau siv los npaj cov qauv nkhaus (y=0.0532x ntxiv rau 0.0339;r=0.9992), thiab TPC raug xam raws li mg gallic acid sib npaug (GAE) ib 100g ntawm hmoov rau qhuav. hnyav hauv paus.

2.4. Kev txiav txim siab ntawm tag nrho cov ntsiab lus Flavonoid. Tag nrho cov ntsiab lus flavonoid (TFC) tau txiav txim siab raws li txoj kev aluminium chloride colorimetric [19]. Ntiv tes millet extract (100ul) tau tov nrog txhuas chloride tov (2 feem pua ​​​​w / v, 100ul), incubated rau 10min ntawm RT, thiab lub absorbance yog ntsuas ntawm 415nm. Quercetin tau siv los npaj cov qauv nkhaus (y=0.0349x-0.2091; r²=0.9974), thiab TFC raug xam raws li mg quercetin sib npaug rau 100g ntawm hmoov ntawm qhov hnyav qhuav.

2.5.Kev txiav txim siab ntawm DPPH Radical Scavenging Activity. Lub peev xwm txhawm rau txhawm rau DPPH radicals tau txiav txim siab raws li txoj kev piav qhia los ntawm Blois [20]. Ntiv tes mil-let extract (50 ul) tau tov nrog 90ul ntawm methanol thiab 60ul ntawm DPPH tov (0.02 feem pua ​​w/v) thiab incubated ntawm RT nyob rau hauv qhov tsaus ntuj rau 10min. Absorbance qhov tseem ceeb ntawm cov qauv (As) thiab tswj (Ac) tau ntsuas ntawm 517nm. Trolox tau siv raws li tus qauv. DPPH radical scavenging kev ua si raws li qhov feem pua ​​​​ inhibition tau suav nrog kev sib npaug (1). Tus nqi ICs tau suav nrog siv cov kab lus teb cov koob tshuaj.

2.6. Kev txiav txim siab ntawm ABTS Cation Radical Scavenging Activity. Lub peev xwm txhawm rau txhawm rau ABTS cation radicals tau txiav txim siab raws li txoj kev piav qhia los ntawm Re et al. [21] nrog qee qhov kev hloov kho. ABTS cation radical tov yog npaj los ntawm incubating 10mg ntawm ABTS nyob rau hauv 2.5 ml ntawm poov tshuaj persulphate ntawm 37 degree C nyob rau hauv qhov tsaus ntuj rau 16h thiab diluting 7 zaug nrog 50mM phosphate buffer saline (pH 7.4). Ntiv tes millet extract (12.5ul) tau tov nrog phosphate buffer saline (147.5ul) thiab freshly npaj ABTS cation radical tov (40ul) thiab incubated ntawm RT nyob rau hauv qhov tsaus ntuj rau 10min. Absorbance qhov tseem ceeb ntawm tus qauv (As) thiab tswj (Ac) tau ntsuas ntawm 734nm. Trolox tau siv raws li tus qauv. ABTS cation radical scavenging kev ua si raws li qhov feem pua ​​​​ inhibition tau suav nrog Equation(2).ICs tus nqi raug suav nrog siv cov kab lus teb.

2.7.Kev txiav txim siab ntawm Oxygen Radical Absorbance Capacity (ORAC). ORAC raug soj ntsuam raws li txoj kev piav qhia los ntawm Ou li al.[22] nrog qee qhov kev hloov kho.Fluorescein (4.8uM) thiab AAPH (40mg / ml) daws tau npaj hauv 75mM phosphate buffer (pH 7.4). Ntiv tes millet extract (10ul) tau tov nrog 40ul ntawm phosphate tsis thiab 100ul ntawm fluorescein thiab incubated ntawm 37 degree rau 10min. AAPH (50ul) tau ntxiv, thiab kev lwj ntawm fluorescein tau ntsuas ntawm kev ua kom zoo thiab emission wavelengths ntawm 494nm thiab 535nm, raws li, ntawm 37 degree rau 35min ntawm kuv min intervals siv lub fluorescence microplate nyeem ntawv. Tro-lox tau siv raws li tus qauv. Thaj chaw nyob rau hauv qhov nkhaus (AUC) tus nqi ntawm cov qauv (AUCs), tswj (AUC), thiab Trolox (AUC-) tau kaw. ORAC tus nqi raug suav nrog Equation(3) qhov twg conc. stands rau concentration. Cov txiaj ntsig tau nthuav tawm raws li mg Trolox sib npaug rau 100g ntawm hmoov ntawm qhov hnyav qhuav.

2.8. Kev txiav txim siab ntawm Ferrous Ion Chelating (FIC) Kev Ua Haujlwm. Lub peev xwm los chelate ferrous ions tau txiav txim siab raws li txoj kev piav qhia los ntawm Carter [23]. Ntiv tes millet extract (100ul) yog tov nrog 1 mM ferrous sulphate tov (20ul) thiab 40ul ntawm distilled dej. Tom qab ntawd, 1 mM ferrozine tov (40ul) tau ntxiv thiab incubated rau 10min ntawm RT. Absorbance qhov tseem ceeb ntawm cov qauv (As) thiab tswj (Ac) tau sau tseg ntawm 562nm. EDTA tau siv los ua tus qauv. FIC kev ua ub no raws li feem pua ​​​​chelation tau suav nrog kev sib npaug (4).

2.9.Kev txiav txim siab ntawm Ferric Txo Antioxidant zog (FRAP). FRAP tau txiav txim siab raws li txoj kev piav qhia los ntawm Benzine thiab Szeto [24] nrog qee qhov kev hloov kho. FRAP reagent tau npaj los ntawm kev sib xyaw 300mM acetate buffer (pH 3.6), 20mM ferric chloride, thiab 10mM TPTZ hauv qhov sib piv ntawm 10: 1: 1 thiab incubating ntawm 37 degree C rau 10min. Ntiv tes millet extract (20ul) tau tov nrog 30ul ntawm acetate tsis thiab 150ul ntawm freshly npaj FRAP reagent, incubated ntawm RT rau 8min, thiab absorbance tau kaw ntawm 600nm. Trolox tau siv los npaj cov qauv nkhaus (y=0.17x ntxiv rau 0.15; 产=1.00), thiab FRAP tau xam raws li mg Trolox sib npaug rau 100g ntawm hmoov ntawm qhov hnyav qhuav.

2.10.Kev txiav txim siab ntawm A5-LOX Inhibitory Activity. A{{0}}LOX inhibitory kev ua haujlwm raug soj ntsuam raws li txoj kev piav qhia los ntawm Tappel [25] nrog qee qhov kev hloov kho. Ntiv tes millet extract (10ul) tau tov nrog 115ul ntawm 100mM sodium phosphate buffer (pH 8.0) thiab 50ul ntawm A5-LOX kua (5000U / ml) thiab incubated ntawm RT rau 10min. Cov tshuaj tiv thaiv tau pib nrog qhov sib ntxiv ntawm 25ul ntawm 0.08mM linoleic acid. Qhov kev hloov ntawm absorbance tau sau tseg ntawm 234nm ntawm 1min ntu rau 10min ntawm RT, thiab qhov siab tshaj plaws tshaj tawm (Vmxk) qhov tseem ceeb ntawm cov qauv (Vmaxs) thiab tswj (Vmaxc) tau kaw. Baicalein tau siv los ua tus qauv. A5-LOX inhibitory kev ua ub no raws li qhov feem pua ​​​​-hnub inhibition tau suav nrog Equation(5).ICs.value tau suav nrog siv cov kab lus teb tshuaj.

2.11.Determination of XO Inhibitory Activity. XO inhibitory kev ua haujlwm tau txiav txim siab raws li txoj kev piav qhia los ntawm Lee li al. [26] nrog qee qhov kev hloov kho. Finger millet extract (10}ul) tau tov nrog 150ul ntawm 50 mM sodium phosphate buffer (pH 7.4) thiab 20ul ntawm XO (0.15U / ml) thiab incubated rau 10min ntawm RT. Cov tshuaj tiv thaiv tau pib nrog qhov sib ntxiv ntawm 20ul ntawm 0.1 mM xanthine. Qhov kev hloov ntawm absorbance tau sau tseg ntawm 295nm ntawm min ntu rau 15min ntawm RT, thiab Vmax qhov tseem ceeb ntawm cov qauv (Vmaxs) thiab tswj (Vmaxc) tau sau tseg. Allopurinol tau siv los ua tus qauv. XO inhibitory kev ua si raws li qhov feem pua ​​​​ inhibition tau suav nrog Equation(6).ICs.value tau suav nrog cov koob tshuaj teb cov duab.

2.12.Kev txiav txim siab ntawm Hyaluronidase Inhibitory Activity. Hyaluronidase inhibitory kev ua haujlwm tau raug soj ntsuam raws li txoj kev piav qhia los ntawm Sahasrabudhe thiab Dedhar [27] nrog qee qhov kev hloov kho. Ntiv tes millet extract (50ul) tau tov nrog 10ul ntawm hyaluronidase (8400U/ml) thiab incubated ntawm 37 degree rau 10min. Lub enzyme tau qhib los ntawm kev ntxiv 20ul ntawm calcium chloride (12.5mM) thiab incubating ntawm 37 degree rau 10min. Cov tshuaj tiv thaiv tau pib nrog qhov sib ntxiv ntawm sodium hyaluronate (50ul) thiab incubated ntawm 37 degree C rau 40min. Tom qab ntawd, 10ul ntawm 0.9 M sodium hydroxide thiab 20ul ntawm 0.2M sodium borate tau ntxiv thiab incubated ntawm 100 degree rau 3min. Tom qab txias rau RT, 50ul ntawm 67mM p-dimethylaminobenzaldehyde tau ntxiv thiab incubated ntawm 37 degree rau 10min. Absorbance qhov tseem ceeb ntawm cov qauv (As) thiab tswj (Ac) tau ntsuas ntawm 585nm. Tannic acid tau siv raws li tus qauv. Hyaluronidase inhibitory kev ua si raws li qhov feem pua ​​​​ inhibition tau suav nrog Equation (7).

2.13.Determination of Oxidative Burst Inhibitory Activities. Anti-inflammatory muaj peev xwm nyob rau hauv cov nqe lus ntawm oxidative burst inhibitory kev ua ub no nyob rau hauv tag nrho cov tib neeg cov ntshav thiab cais polymorphonuclear neutrophils tau txiav txim siv lub luminol enhanced chemiluminescence assay [28, 29] nrog rau ib co kev hloov. Kev sim tau ua nyob rau ntawm Dr. Panjwani's Center for Molecular Medicine and Drug Research, International Center for Chemical and Biological Sciences, University of Karachi, Pakistan, thiab lub koom haum muaj kev ncaj ncees clearance rau kev kawm ntawm tib neeg cov ntshav los ntawm lub ywj siab ethics committee, ICCBS, UoK . Tsis yog:ICCB-S/IEC-008-BC-2015/Protocol/1.0.

2.13.1. Kev txiav txim siab ntawm Oxidative Burst Inhibitory Activity nyob rau hauv tag nrho tib neeg cov ntshav. Tib neeg cov ntshav (1ml) tau txais aseptically sau los ntawm cov hlab ntsha mus rau hauv lub raj heparinized los ntawm cov neeg ua haujlwm pabcuam noj qab haus huv tsis haus luam yeeb, uas tsis noj tshuaj lossis tshuaj ntxiv rau ntau tshaj ib lub lis piam, thiab diluted (1:20 dilution) nrog HBSs ntxiv * (Hanks Balanced Cov tshuaj ntsev, muaj calcium thiab magnesium). Ntiv tes millet extract (25ul) yog tov nrog diluted tag nrho cov ntshav (25 ul) thiab incubated ntawm 37 degree C rau 15 feeb. Tom qab ntawd, 25ul ntawm serum opsonized zymosan thiab 25ul ntawm luminol tau ntxiv. Lub oxidative tawg ROS ntau lawm tau saib xyuas rau 50min siv lub luminometer nrog rov kuaj dua ntawm 30s ntu thiab 1 s cov ntsiab lus ntsuas lub sijhawm. Ibuprofen tau siv los ua tus qauv tshuaj. Qhov feem pua ​​​​ inhibition tau suav nrog Equation (8).

qhov twg RLUis lub luminometer nyeem nyob rau hauv cov nqe lus ntawm cov txheeb ze lub teeb units (RLU) rau kev tswj thiab RLU yog lub luminometer nyeem nyob rau hauv RLU rau cov qauv. Tus nqi ICs tau suav nrog siv cov kab lus teb cov koob tshuaj.

2.13.2. Kev txiav txim siab ntawm Oxidative Burst Inhibitory Activity hauv Isolated Polymorphonuclear Neutrophils. Tag nrho tib neeg cov ntshav (10 ml) tau sib xyaw nrog qhov sib npaug ntawm HBSS (Hanks Balanced Salt Solution, tsis muaj calcium thiab magnesium) thiab lymphocyte cais nruab nrab (LSM) thiab tso cai rau erythrocytes nyob rau 30 min ntawm RT. Tom qab ntawd, cov ntshav tau muab tso rau hauv LSM kom zoo thiab centrifuged ntawm 400 × g rau 20min ntawm RT. Tus supernatant raug tshem tawm; Cov qe ntshav liab nyob rau hauv qhov tshwm sim pellet tau lysed los ntawm kev sib tov nrog dej txias deionized, tov nrog txias HBSS7, thiab centrifuged ntawm 300 × g rau 10min ntawm 4 degree. Cov pellet tau ntxuav ob zaug, thiab qhov ua tau zoo ntawm cov txiaj ntsig ntawm cov tshuaj polymorphonuclear neutrophils raug tshuaj xyuas siv Trypan blue exclusion method. Cov concentration ntawm cov hlwb raug kho rau 1 × 10 degree hlwb / ml. cais polymorphonuclear neutrophils (25 山l) tau tov nrog ntiv tes millet extract (25ul) thiab incubated rau 15 min ntawm 37 degree. Tom qab ntawd, 25ul ntawm serum opsonized zymosan thiab 25ul ntawm luminol tau ntxiv, thiab oxidative tawg ROS ntau lawm tau saib xyuas rau 50 min siv lub luminometer nrog rov ua dua ntawm 30s ntu thiab ls cov ntsiab lus ntsuas lub sijhawm. Ibuprofen tau siv los ua tus qauv tshuaj. Qhov feem pua ​​​​ inhibition tau suav nrog Equation(8).

2.14. Kev txheeb cais. Cov ntaub ntawv ntawm txhua qhov kev sim tau raug txheeb xyuas los ntawm kev siv IBM SPSS Statistics (version 20) software, thiab cov txiaj ntsig tau qhia tias txhais tau tias ± tus qauv (SE). Kev txheeb cais tseem ceeb tau teeb tsa ntawm 95 feem pua ​​​​kev ntseeg siab. Ib txoj kev tsom xam ntawm qhov sib txawv (ANOVA) thiab Tukey qhov kev sim tau siv los txiav txim siab qhov sib txawv ntawm ntau yam thiab rho tawm. Pearson's correlation coefficient tau siv rau kev txheeb xyuas kev sib raug zoo.

Classification ntawm Inflamation

Mob o: feem ntau yog tshwm sim los ntawm liab, o, mob, thiab lwm yam, uas yog, o feem ntau tshwm sim los ntawm cov tshuaj tiv thaiv ntawm vascular system.

Mob o: feem ntau yog qee cov kab mob inflammatory, xws li mob gastroenteritis, kab mob siab ntev, rov tshwm sim gynecological o thiab lwm yam kab mob.

Ua rau Inflamation

Txhua yam uas tuaj yeem ua rau cov ntaub so ntswg puas tuaj yeem ua rau o, uas yog, inflammatory yam tuaj yeem muab faib ua pawg hauv qab no:

(1) Cov khoom siv roj ntsha

Cov kab mob, kab mob, mycoplasmas, fungi, spirochetes, thiab parasites yog feem ntau ua rau mob. Kev mob tshwm sim los ntawm cov kab mob lom neeg kuj hu ua kab mob. Exotoxins thiab endotoxins tsim los ntawm cov kab mob tuaj yeem ua rau cov ntaub so ntswg ncaj qha; Cov kab mob sib kis hauv cov kab mob kis ua rau cov cell necrosis; Qee cov tshuaj tiv thaiv kab mob ua rau cov ntaub so ntswg los ntawm kev tiv thaiv kab mob tom qab kis kab mob, xws li kab mob parasitic thiab tuberculosis.

2) Cov yam ntxwv ntawm lub cev

Qhov kub thiab txias, tsis tshua muaj kub, cov khoom siv hluav taws xob, hluav taws xob ultraviolet, thiab lwm yam, thiab kev puas tsuaj rau txhua yam.

(3) Chemical yam

Exogenous tshuaj xws li muaj zog acid, muaj zog alkali, thiab turpentine. Endogenous toxic tshuaj xws li decomposition khoom ntawm necrotic ntaub so ntswg thiab metabolites xws li urea accumulated nyob rau hauv lub cev nyob rau hauv tej yam pathological mob.

(4) Lub cev txawv teb chaws

Lub cev txawv teb chaws nkag mus rau tib neeg lub cev los ntawm ntau txoj hau kev, xws li ntau yam hlau, ntoo khib nyiab, plua plav hauv huab cua, kev phais phais, thiab lwm yam, tuaj yeem ua rau muaj kev sib txawv ntawm cov lus teb vim yog lawv qhov sib txawv antigenicity.

(5) Cov ntaub so ntswg necrotic

Ua rau xws li ischemia lossis hypoxia tuaj yeem ua rau cov ntaub so ntswg necrosis, uas yog qhov muaj peev xwm ua rau mob. Congestive hemorrhagic bands thiab infiltration ntawm inflammatory hlwb ntawm cov npoo ntawm tshiab infarcts yog ob qho tib si tshwm sim ntawm o.

(6) Kev ua xua

Thaum lub cev tiv thaiv kab mob tsis zoo, nws tuaj yeem ua rau lub cev tsis tsim nyog lossis ntau dhau, ua rau cov ntaub so ntswg thiab cell puas thiab ua rau mob. Xws li kev tsis haum rhinitis, colitis, urticaria, tuberculosis, glomerulonephritis, thiab lwm yam kab mob.

Cov tshuaj tua kab mob yog cov kab mob bactericidal thiab tiv thaiv kab mob, kev siv mus ntev yuav ua kom lub cev muaj zog ntawm tib neeg lub cev, tua cov kab mob zoo, ua rau lub plab zom mov tsis txaus, ua rau muaj ntau yam kab mob hauv plab hnyuv thiab tsis zoo, thiab ua rau cov kab mob thib ob, txo lub cev. tsis kam. Tsis tas li ntawd, cov tshuaj tua kab mob feem ntau yog metabolized los ntawm lub siab thiab lub raum, thiab kev tsim txom ntawm cov tshuaj tua kab mob feem ntau yuav ua rau lub siab thiab lub raum ua haujlwm. Koj tuaj yeem xyuam xim rau cov lus "siv ceev faj rau cov neeg mob uas muaj lub siab tsis zoo thiab lub raum ua haujlwm" ntawm cov lus qhia ntawm txhua yam tshuaj tiv thaiv. Tsis tas li ntawd, cov tshuaj tua kab mob tuaj yeem ua rau muaj kev tsis haum tshuaj rau cov tshuaj. Nyob rau hauv xyoo tas los no, qhov tshwm sim siab ntawm kev tsis haum rhinitis thiab kev ua xua mob hawb pob yog cuam tshuam nrog kev tsim txom ntawm cov tshuaj tua kab mob.

Kev sim tau pom tias cov extract ntawm Cistanche pipiensis nplua nuj nyob rau hauv echinacoside muaj kev txhim kho zoo ntawm colitis; Cistanche glycoside K thiab piposide B muaj qhov cuam tshuam zoo rau kev tso tawm ntawm nitric oxide hauv hlwb, qhia tias nws cov txiaj ntsig zoo los tiv thaiv.Cistancheyog cov khoom siv tshuaj suav nrog tib lub hauv paus chiv keeb xws li tshuaj thiab zaub mov. Nws yog ib qho ntawm kaum suav tshuaj ntsuab tshuaj ntsuab hauv kuv lub tebchaws. Nws yog ib qho khoom muaj nqis los ntawm qhov tob ntawm cov suab puam thiab lub ntuj tonic. Cistanche Cistanche muaj keeb kwm tshuaj muaj ze li ntawm 2, 000 xyoo. Nws tau sau thawj zaug hauv "Shen Nong's Materia Medica". Xyoo 2005, Cistanche tau suav nrog hauv "Pharmacopoeia ntawm Cov Neeg Sawv Cev ntawm Tuam Tshoj" ua cov khoom siv tshuaj tiag tiag. Cistanche tau ua ib qho zoo tonic txij thaum ub los, tsis muaj cov ntaub ntawv toxicity, uas ua pov thawj tag nrho nws txoj kev nyab xeeb.