Lub Mechanism ntawm Cistanche Deserticola Txhawb Txoj hnyuv ua haujlwm

Yuan Gao, Chuanjie Zong, Fen Liu, Lei Fang, Runlan Cai, Yue Shi, Xi Chen, Yun Qi

Abstract

Phenylethanoidglycosides(PhGs), chav kawm ntawm polyphenolic tebchaw, suav hais tias yog ib qho ntawm cov khoom siv bioactive tseem ceeb ntawmCistanchedeserticolaYC Ma (CD), uas nws cov extract yog qhov ncauj siv hauv tshuaj suav tshuaj. Txawm hais tias yav dhau los kev tshawb fawb pharmacological tau tshaj tawm tias PhGs ua haujlwm ntau yam, lawv cov kab mob hauv plab hnyuv tsis tau qhia meej. Hauv txoj kev tshawb no, peb tau tshawb xyuas cov hnyuv permeability ntawm PhG-nplua nuj extract (PRE) los ntawmCistancheDeserticolaraws li kev sib xyaw ua ke hauv Caco-2 cell monolayer qauv siv bioassay system. Cov txiaj ntsig tau pom tias PRE feem ntau yog thauj los ntawm qhov tsis zoo ntawm qhov nqus tsis zoo ntawm qhov kev tawm mus rau hauv qhov concentration gradient yam tsis muaj efflux, uas muab cov tshuaj pharmacokinetic rau kev siv tshuaj kho mob ntawm PhGs.Cistanche Deserticola. Peb kuj tau txiav txim siab txoj hnyuv permeability ntawm peb qhov lojPhGs[Acteoside (AC), isoacteoside (IS), thiab echinacoside (EC)]by HLPC. Tsis tas li ntawd, peb tau tsim ib txoj kev tshawb nrhiav HPLC-fluorescence tshiab los txiav txim siab qhov flux ntawm AC thiab IS. Raws li xav tau, cov yam ntxwv thauj ntawm pebPhGsyog raws li cov PRE, qhia tias tam sim no bioassay system yog tsim nyog thiab txhim khu kev qha rau kev ntsuam xyuas ntawm cov yam ntxwv thauj ntawm active ingredient pawg (AIG) nyob rau hauv PRE. Ntxiv mus, qhov system no kuj yuav tsim nyog rau lwm cov nroj tsuag rho tawm uas tsim nyog bioactivity.

Hu rau:joanna.jia@wecistanche.com/ WhatsApp: 008618081934791

Cistanche deserticola muaj ntau yam teebmeem, nyem qhov no kom paub ntau ntxiv

Taw qhia

Caco-2 kab ntawm tes, uas tau muab los ntawm tib neeg txoj hnyuv adenocarcinomas, pom cov yam ntxwv zoo li enterocyte. Raws li ib txwm muaj, Caco-2 cov hlwb spontaneously sib txawv ntawm cov hlwb paub tab thiab tsim cov monolayers tsis zoo [1]. Cov hlwb uas nyob ib sab ua raws li kev sib tshuam nruj tsim nyob rau sab apical ntawm lub monolayer, uas tuaj yeem cais cov tshuaj uas tsis muaj zog thiab nquag thauj cov tshuaj hla cov txheej txheem epithelial [2]. Vim yog cov morphological thiab biochemical zoo sib xws rau cov enterocytes ib txwm, Caco-2 cell monolayers ua ib tug zoo-tsim nyob rau hauv vitro qauv rau txoj kev kawm ntawm lub plab hnyuv muaj peev xwm nqus thiab thauj cov yam ntxwv ntawm cov tshuaj [3, 4].

Hauv kev sib piv rau cov tshuaj, cov nroj tsuag rho tawm (PE) yog cov khoom sib xyaw uas nws cov kev ua haujlwm lom neeg thiab cov khoom siv nquag feem ntau tsis pom zoo [5]. Ntxiv mus, cov khoom siv hauv plab hnyuv ntawm PE, tsis zoo li cov khoom ntawm nws cov khoom, muaj feem cuam tshuam nrog kev siv tshuaj kho mob. Kev ntsuas Flux rau cov qauv ntsuas thoob plaws Caco-2 cell monolayer feem ntau koom nrog cov txheej txheem tshuaj, xws li HPLC, LC/MS, thiab lwm yam. Txawm hais tias cov txheej txheem no yog cov cuab yeej muaj zog, lawv nyuaj, siv sij hawm, kim, thiab qee zaus xav tau cov cuab yeej ntse. Qhov tseem ceeb tshaj, tsis yog ib leeg lossis ib pawg neeg tsawg tuaj yeem cuam tshuam PE tag nrho. Yog li, ib txoj hauv kev tshiab ywj pheej ntawm kev txiav txim siab ntawm cov neeg koom nrog yuav tsum tau tsim los txheeb xyuas thiab ntsuas cov yam ntxwv ntawm kev thauj mus los ntawm PE.

CistanchedeserticolaYC Ma (CD), cov nroj tsuag holoparasitic, yog ib hom tshuaj suav tshuaj uas siv los kho lub raum tsis muaj zog, lub cev tsis muaj zog, thiab cem quav, thiab cov kev siv no tau raug sau tseg hauv Suav Pharmacopoeia [6]. Phenylethanoid glycosides (PhGs), suav nrog echinacoside (EC), acteoside (AC), isoacteoside (IS), thiab lwm yam, yog chav kawm ntawm polyphenolic compounds [7]. Lawv suav hais tias yog ib qho ntawm cov tseem ceeb bioactive cov khoom ntawmCistanchelaj [8]. Pharmacological kev tshawb fawb tau pom tias bioactivity ntawmPhGsmuaj ntau haiv neeg thiab suav nrog cov tshuaj tiv thaiv oxidative [9], tiv thaiv qaug zog [10], hepatoprotective [11], immunomodulatory [12], anti-inflammatory [7, 13] thiab cov teebmeem neuroprotective [14]. Txawm li cas los xij, txoj hnyuv thauj yam ntxwv ntawmPhGstsis tau tshawb nrhiav. Hauv txoj kev tshawb no, peb tau tshawb xyuas cov hnyuv permeability ntawm PhG-nplua nuj extract (PRE) los ntawm CD raws li ib qho kev sib koom ua ke thiab permeability ntawm peb loj PhGs (AC, IS, thiab EC) nyob rau hauv txawv Caco-2 hlwb. Peb cov txiaj ntsig tau qhia tias PRE feem ntau yog thauj mus los ntawm qhov tsis zoo ntawm qhov nqus tsis zoo ntawm cov pa tawm mus rau qhov concentration gradient yam tsis muaj efflux, uas muab cov tshuaj pharmacokinetic rau kev kho mob ntawm PhGs hauv CD.

Cov ntaub ntawv thiab cov txheej txheem

Khoom siv

The human intestinal Caco-2 cell line was obtained from the American Type Culture Collection (ATCC, Rockville, MD, USA). AC, IS and EC (>98 feem pua) tau yuav los ntawm Yuav Tsum Biotechnology Co. (Chengdu, Suav). Dulbecco's modified Eagle's medium (DMEM), fetal bovine serum (FBS), thiab non-essential amino acids (NEAA) tau tsim los ntawm Gibco BRL (Grand Island, NY, USA). 6-zoo TranswellTM daim hlau (insert membrane loj hlob cheeb tsam 4.67 cm2 ) tau los ntawm Corning (Costar) Inc. (Tewksbury, MA, USA). Rat-tail collagen tau txais los ntawm Sigma-Aldrich (St. Louis, MO, USA). Tag nrho cov tshuaj reagents thiab tshuaj rau HPLC tsom xam yog qib analytical.

Kev npaj PRE los ntawm Cistanche deserticola

Cov khoom cua-dried CD yog hmoov thiab muab rho tawm los ntawm percolation nrog 70 feem pua ​​​​ethanol. PhG-nplua nuj feem tau npaj raws li tau piav qhia yav dhau los [10] thiab muab rho tawm nrog dej-saturated n-butyl cawv. Cov cawv extract tau concentrated thiab qhuav nyob rau hauv lub siab txo. Macroporous resin-UV spectrophotometry [15] ntsuas PhG cov ntsiab lus ntawm 78.4 feem pua. Cov qauv kawg sawv cev rau 1.75 feem pua ​​​​ntawm cov khoom siv raw los ntawm qhov hnyav qhuav. Cov qauv tau muab khaws cia ntawm -20 degree kom txog thaum siv ntxiv.

Kev txiav txim siab ntawm AC, IS, thiab EC los ntawm HPLC

Shimadzu HPLC system nruab nrog LC daws software tau siv los ntsuas cov ntsiab lus ntawm AC, IS, thiab EC hauv PRE. Ib qho rov qab-theem Intersil C18 kem (4.6 mm × 250 mm, 5 μm) tau siv thiab khaws cia ntawm chav tsev kub. Cov theem mobile yog acetonitrile thiab dej uas muaj 0.1 feem pua ​​​​phosphoric acid (v/v) nrog gradient elution (Table 1) ntawm tus nqi ntws ntawm 1.0 ml / min. UV spectrophotometer ntes tau teem rau 334 nm. Txhawm rau txiav txim siab qhov flux ntawm AC thiab IS, peb tau tsim ib txoj hauv kev tshiab HPLC-fluorescence detection (HPLC-FLD). Tom qab kev soj ntsuam cov qauv thiab cov fluorescence wavelength scanning (cov ntaub ntawv tsis qhia), peb tau txais qhov zoo tshaj plaws fluorescence nrhiav pom tej yam kev mob rau AC (Ex: 338 nm, Em: 448 nm) thiab IS (Ex: 320 nm, Em: 434 nm)

HPLC txoj kev tshuaj ntsuam xyuas tau raug siv tau siv cov yam ntxwv ua tau zoo hauv qab no: kev ruaj ntseg, linearity, rhiab heev, precision (intra- thiab inter-day variability), thiab raug. Cov analytes nyob rau hauv lub thauj tsis tau muab khaws cia ntawm 25 degree nyob rau hauv qhov tsaus ntuj rau 24 teev, thiab lawv stability yog ntsuas. Cov analytes tau ruaj khov nyob rau hauv lub xub ntiag ntawm vitamin C (0.4 feem pua) vim hais tias tus txheeb ze tus qauv sib txawv (RSD) qhov tseem ceeb nyob rau hauv lub ncov cheeb tsam yog <3.5 feem="" pua.="" cov="" kab="" sib="" npaug="" sib="" npaug,="" sib="" txuas="" coefficient,="" ntau="" yam="" ntawm="" linearity,="" lod,="" thiab="" loq="" rau="" ac,="" is,="" thiab="" ec="" yog="" qhia="" hauv="" table="" 2.="" lods="" (18–3{20}}="" nm)="" thiab="" loqs="" (60–100="" nm)="" qhov="" tseem="" ceeb="" qhia="" tau="" hais="" tias="" hplc-fld="" thiab="" hplc-uv="" txoj="" kev="" muaj="" kev="" nkag="" siab="" zoo.="" rsd="" qhov="" tseem="" ceeb="" uas="" qhia="" txog="" qhov="" tseeb="" ntawm="" txoj="" kev="" yog="" tag="" nrho=""><2 feem="" pua="" ​​​​rau="" qhov="" sib="" txawv="" hauv="" nruab="" nrab="" hnub,="" qhia="" qhov="" tseeb="" zoo.="" rau="" qhov="" kev="" ntsuam="" xyuas="" rov="" qab,="" ac,="" is,="" thiab="" ec="" tau="" ntxiv="" rau="" cov="" khoom="" thauj="" tsis="" zoo="" kom="" ua="" tiav="" peb="" qib="" siab="" (siab,="" nruab="" nrab,="" thiab="" qis).="" cov="" txiaj="" ntsig="" rov="" qab="" los="" ntawm="" cov="" txheej="" txheem="" rau="" cov="" tshuaj="" ntsuam="" xyuas="" tau="" sau="" tseg="" hauv="" table="" 3.="" qhov="" nruab="" nrab="" qhov="" rov="" qab="" tau="" yog="" li="" ntawm="" 90.0="" feem="" pua="" ​​​​rau="" 96.4="" feem="" pua,="" qhia="" qhov="" tseeb="" zoo.="" hauv="" kev="" xaus,="" cov="" txheej="" txheem="" hplc="" tau="" tsim="" muaj="" kev="" txaus="" siab="" rau="" kev="" hais="" txog="" linearity,="" rhiab="" heev,="" precision,="" thiab="" raug="" rau="" qhov="" kom="" muaj="" nuj="" nqis="" ntawm="" ac,="" is,="" thiab="" ec="" hauv="" kev="" thauj="" khoom="">

Kev txiav txim siab ntawm PRE los ntawm bioassay system

Lub bioassay (tag nrho cov tshuaj tiv thaiv oxidative muaj peev xwm) tau ua raws li FRAP (ferric txo / antioxidant zog) kev soj ntsuam peb yav dhau los tau piav qhia [16] thiab siv tau raws li cov kab sib txawv thiab qhov tseeb (intra- thiab inter-hnub hloov pauv). Lub linearity ntawm txoj kev bioassay tau txiav txim siab raws li cov calibration curves. Qhov ntau npaum li cas ntawm linearity yog 0.0625 − 40 ug/ml (y=0.0258x ntxiv rau 0.0968, R2=0.996). Qhov tseeb ntawm txoj kev tsim bioassay tau txiav txim siab nyob rau hauv cov nqe lus ntawm intra- thiab inter-hnub variability rau kev soj ntsuam ntawm PRE (10.0 ug / ml). Txoj kev rov ua dua ib hnub ntawm txoj kev tau txiav txim siab raws li tsib qhov kev txiav txim sib law liag rau tib hnub. Qhov nruab nrab hnub rov ua tau raug ntsuas raws li tsib qhov kev txiav txim sib law liag rau peb hnub sib txawv. RSD qhov tseem ceeb rau qhov tseeb ntawm txoj kev yog 1.014 feem pua ​​​​thiab 4.72 feem pua ​​​​rau intra- thiab inter-hnub sib txawv, raws li, qhia tau hais tias zoo precision. Yog li, txoj kev tsim bioassay yog qhov txaus siab rau kev sib tw linearity, precision, thiab raug rau qhov kom muaj nuj nqis ntawm PRE hauv kev thauj tsis tau.

Cell kab lis kev cai

Caco{{0}} cov hlwb raug coj los ntawm 37 degree thiab 95 feem pua ​​​​ntawm cov av noo nyob rau hauv ib qho chaw uas muaj 5 feem pua ​​CO2 thiab nruab nrab muaj DMEM, 10 feem pua ​​FBS, 1 feem pua ​​​​NEAA, 1 feem pua ​​L-glutamine, penicillin ( 100 U / ml), thiab streptomycin (100 ug / ml). Qhov nruab nrab tau hloov pauv txhua hnub thaum lub sijhawm kev loj hlob ntawm tes thiab kev sib txawv. Cov hlwb tau loj hlob hauv 75-cm2 lub khob yas thiab sau txhua 3-5 hnub nrog 0.05 feem pua ​​​​EDTA-trypsin. Rau kev thauj mus los, cov hlwb tau cog ntawm qhov ntom ntawm 1 × 105 hlwb / cm2 ntawm Transwell inserts coated nrog collagen. Kwv yees li 21 hnub tom qab cog, cov monolayers tau siv rau kev sim thauj. Lawv txoj kev ncaj ncees tau txiav txim siab los ntawm kev ntsuas qhov trans-epithelial hluav taws xob tsis kam (TEER) hla cov monolayers nrog EVOM nruab nrog ENDOHM-SNAP (World Precision Instruments, Inc., USA). TEER qhov tseem ceeb ntawm Caco-2 cell monolayer xav tau tshaj 500 O∙cm2.

Kev sim thauj mus los

Cov monolayer tau ntxuav nrog thauj tsis muaj (P-buffer muaj 10 mM HEPES, 1 mM sodium pyruvate, 10 mM qabzib, 3 mM CaCl2 thiab 145 mM NaCl, pH 7.4) thiab tom qab ntawd ua ntej incubated rau 20 min ntawm 37 degree. Tom qab tshem tawm cov kev thauj mus los, cov khoom thauj tshiab tsis muaj cov qauv kuaj tau muab ntxiv rau hauv apical (AP) chamber (1.5 ml) hauv AP mus rau basolateral (BL) cov kev tshawb fawb qhia lossis BL chamber (2.5 ml) hauv BL rau AP cov kev tshawb fawb. Rau AC, IS thiab EC kev soj ntsuam uas siv HPLC, ib qho aliquot (300 ul) raug tshem tawm ntawm txhua chav txais khoom ntawm lub sijhawm sib txawv (30, 60, 90, thiab 120 min). Lub receiver chamber tau ntxiv nrog tib lub ntim ntawm cov tshiab preheated (37 degree) thauj tsis tau tom qab txhua qhov kev kuaj. Cov qauv sau tau muab khaws cia ntawm -20 degree rau kev siv ntxiv. Rau kev txiav txim siab PRE siv bioassay, tsuas yog cov qauv ntawm 120 min raug coj mus. Vim qhov tsis ruaj khov ntawm PRE, AC, IS, thiab EC, 0.4 feem pua ​​(w/v) vitamin C tau ntxiv kom ruaj khov cov qauv txhawm rau txiav txim siab AC, IS, thiab EC cov ntsiab lus los ntawm HPLC, thiab cov qauv rau PRE bioassay tau kuaj tam sim tom qab kuaj. Qhov pom meej permeability coefficient (Papp lossis 'Papp) tus nqi ntawm txhua tus qauv raug xam.

Cov ntaub ntawv tsom xam

Cov txiaj ntsig Papp hauv AP BL lossis BLAP cov lus qhia ntawm AC, IS thiab EC tau suav ua raws li qhov sib npaug hauv qab no: Papp {{0}} (4Q/4t)/(AC0), qhov twg Papp yog qhov pom meej permeability coefficient (cm / s) txiav txim los ntawm HPLC. (4Q / 4t) yog tus nqi ntawm cov tsos ntawm cov tshuaj ntsuam xyuas (AC, IS, lossis EC) ntawm sab txais (μmol / s); A yog thaj tsam ntawm qhov ntxig (cm2); C0 yog qhov kev sim thawj zaug sib xyaw ua ke ntawm tus neeg pub dawb (μmol / ml). Tshwj xeeb, chav tsev loj "ug" tau siv tsis yog "μmol" thaum 'Papp qhov tseem ceeb raug xam. Cov ntaub ntawv yog qhia raws li qhov txhais tau tias ± SD.

Cov txiaj ntsig

Kev sib xyaw ntawm Acteoside, Isoacteoside, thiab Echinacoside hauv PRE los ntawm Cistanche Deserticola

Vim tias AC, IS thiab EC suav hais tias yog qhov tseem ceeb ntawm PhGs bioactive ntawm Cistanche hom [8], peb tau txheeb xyuas lawv cov ntsiab lus hauv PRE ntawm HPLC-UV. Tus neeg sawv cev chromatogram tau qhia hauv daim duab 1. Kev txheeb xyuas ntawm cov khoom siv no yog ua raws li kev sib piv lub sijhawm khaws cia thiab UV spectrum nrog cov qauv tseeb ntawm lub wavelength ntawm 334 nm. Cov ntsiab lus ntawm AC, IS thiab EC hauv PRE yog 26.60 feem pua, 1.84 feem pua, thiab 32.83 feem pua. Yog li, peb lub tebchaw no suav nrog 61.27 feem pua ​​​​ntawm PRE; Ntxiv mus, cov txiaj ntsig saum toj no qhia tau tias PhG cov ntsiab lus hauv PRE yog 78.4 feem pua. Yog li, AC, IS thiab EC suav txog kwv yees li 80 feem pua ​​​​ntawm PhGs hauv PRE.

Tag nrho cov tshuaj tiv thaiv oxidative muaj peev xwm ntawm PRE, AC, IS, thiab EC

Kev tshawb fawb txog kev tiv thaiv oxidative ntawm PhGs los ntawm CD tau tshaj tawm [9], thiab AC, IS thiab EC suav txog kwv yees li 80 feem pua ​​​​ntawm PhGs hauv PRE. Yog li, tag nrho cov tshuaj tiv thaiv oxidative muaj peev xwm ntawm PRE, AC, IS thiab EC tau soj ntsuam thiab tau qhia siv FRAP qhov tseem ceeb (× 10-6 mmol). Raws li pom hauv daim duab 2, thaum tus nqi FRAP yog 8 × 10-6 mmol, qhov kawg ntawm PRE, AC, IS thiab EC yog 6.20 ug / ml, 3.14 ug / ml, 22.92 ug / ml, thiab 4.89 ug / ml, ntsig txog, qhia tias tag nrho cov tshuaj tiv thaiv oxidative muaj peev xwm ntawm plaub cov qauv no suav nrog hauv qab no: AC > EC > PRE > IS. Lub bioactivity ntawm PRE yog ntaus nqi rau nws cov active ingredient pawg (AIG). Txog li ntawm 60 feem pua ​​​​ntawm PRE, AC thiab EC tau pom muaj zog tag nrho cov kev tiv thaiv oxidative ntau dua li PRE thiab IS. Vim tias IS (1.84 feem pua) yog ib feem me me ntawm PRE, lwm yam khoom siv, xws li cov tsis muaj zog tiv thaiv oxidative IS, kuj tseem ua kom pom tseeb hauv PRE. Kuj ceeb tias, tag nrho cov tshuaj tiv thaiv oxidative sib txawv los ntawm ze li ntawm 7-fold ntawm AC thiab nws isomer IS, qhia tsis tau tsuas yog tus naj npawb ntawm phenolic hydroxyl pawg [9] tab sis kuj txoj hauj lwm ntawm phenolic hydroxyl pab pawg nyob rau hauv lub molecule cuam tshuam lub anti. - oxidative nyhuv.

Kev lees paub ntawm Caco-2 monolayers

Txhawm rau kom siv tau cov Caco-2 cell monolayer system, Papp qhov tseem ceeb ntawm propranolol (ib qho zoo-trans ported marker) thiab Lucifer daj (ib qho cim thauj tsis zoo) los ntawm AP mus rau BL hla Caco-2 monolayers tau txiav txim siab raws li 1.51 × 10–5 cm / s thiab 2.8 × 10–7 cm / s, ntsig txog, thiab cov txiaj ntsig tau pom zoo nrog cov ntawv tshaj tawm yav dhau los [17, 18]. Qhov kev ntsuam xyuas alkaline phosphatase kuj tau lees paub tias Caco-2 cell monolayers tau zoo piv rau cov hnyuv me me epithelium [19]

Lub permeability ntawm Acteoside, Isoacteoside, thiab Echinacoside

In general, the Papp values of well-absorbed drugs were high (>1.< 1.0="" ×="" 10–6="" cm/s)="" [3].="" as="" shown="" in="" table="" 4,="" the="" papp="" values="" of="" ac,="" is="" and="" ec="" was="" nearly="" on="" the="" order="" of="" 10–7="" cm/s,="" indicating="" that="" these="" compounds="" were="" poorly="" permeable.="" furthermore,="" efflux="" or="" active="" transport="" were="" not="" observed="" because="" the="" ratios="" of="" papp="" bl="" to="" ap="" papp="" ap="" to="" bl="" for="" ac,="" is="" and="" ec="" was="" between="" 0.90="" ~="" 1.55,="" and="" the="" criterion="" of="" net="" efflux="" proposed="" by="" the="" fda="" guidance="" is="" a="" ratio="" less="" than="" 2="" [20].="" based="" on="" the="" kinetic="" curves="" presented="" in="" fig.="" 3,="" the="" bidirectional="" transport="" percentages="" of="" ac,="" is="" and="" ec="" at="" 200="" μm="" increased="" linearly="" with="" time.="" the="" transport="" rate="" (tr)="" values="" of="" the="" three="" compounds="" increased="" linearly="" in="" both="" directions="" between="" approximately="" 100="" and="" 300="" μm="" (fig.="" 4).="" these="" results="" indicate="" that="" the="" main="" transport="" mechanism="" of="" ac,="" is="" and="" ec="" is="" also="" passive="">

Lub permeability ntawm PRE

Cov txiaj ntsig saum toj no qhia tias tag nrho cov tshuaj tiv thaiv oxidative muaj peev xwm ntawm PRE yog tsawg kawg yog vim 61.27 feem pua ​​​​ntawm AIG los ntawm PRE. Yog li, peb tau soj ntsuam TR ntawm PRE raws li nws cov concentration-zoo nkhaus ntawm tag nrho cov antioxidative muaj peev xwm thiab xam cov 'Papp qhov tseem ceeb nyob rau hauv thiaj li yuav cuam tshuam cov yam ntxwv thauj ntawm PRE. Qhov 'Papp qhov tseem ceeb ntawm PRE rau AP!BL thiab BL!AP yog (2.16 ± 0.26) × 10–7 cm/s thiab (3.13 ± 0.29) × 10–7 cm/ s, ntsig txog, qhia tias qhov sib xyaw no tsis zoo permeable. Tsis tas li ntawd, efflux lossis active thauj tsis tau tshwm sim vim qhov piv ntawm 'Papp BL rau AP / 'Papp AP rau BL rau PRE yog 1.45 [20]. Ntxiv mus, qhov bidirectional TR ntawm PRE nce linearly ntawm kwv yees li 300 thiab 900 ug / ml (Fig. 5). Qhov tsis muaj qhov kev nyiam ntawm qhov kev tshwm sim qhia tau hais tias passive diffusion yog lub ntsiab kev thauj mus los ntawm PRE.

Kev sib tham

Piv nrog rau cov khoom siv tshuaj purified heev, PE feem ntau yog cov khoom sib xyaw uas muaj ntau pua cov khoom sib txawv nrog cov khoom sib txawv ntawm lub cev. Yog li ntawd, PE exerts systematic, multitarget, thiab multichannel synergistic nkaus vim lawv complex AIG [21], uas cuam tshuam rau kev tsom xam ntawm cov yam ntxwv thauj ntawm PE. Txhawm rau ntsuas cov khoom thauj khoom ntawm PE ntau qhov kev tshawb fawb, qee cov kws tshawb fawb tau txheeb xyuas ntau yam, tsis yog ib feem, hauv PE [22–24]. Txawm li cas los xij, qee qhov txwv tsis pub muaj peev xwm cuam tshuam txog PE tag nrho. Txawm li cas los xij, kev txiav txim siab tag nrho cov khoom hauv PE yog tsis yooj yim sua. Ntxiv mus, yog tias muaj ntau yam khoom hauv PE tso saib cov yam ntxwv sib txawv ntawm kev thauj mus los, cov yam ntxwv ntawm kev thauj mus los ntawm PE yuav nyuaj rau txheeb xyuas.

Hauv xyoo 1990, bioassay systems tau siv los ntsuas TR ntawm cov tshuaj tua kab mob [25]. Los ntawm 2005, Eguchi thiab nws cov npoj yaig [26] tau soj ntsuam cov kev tiv thaiv oxidative ntawm cov zaub ntug hauv paus extract los ntawm kev siv BL xov xwm los ntawm txawv Caco-2 hlwb; txoj hauv kev no yog qhov tsim nyog los xav txog hauv vivo cov xwm txheej.

Raws li daim ntawv tshaj tawm yav dhau los ntawm kev ua haujlwm ntawm PhGs [9] thiab peb cov txiaj ntsig (Daim duab 2), TR ntawm AIG hauv PRE tuaj yeem raug soj ntsuam los ntawm kev txiav txim siab tag nrho cov tshuaj tiv thaiv kab mob ntawm qhov nruab nrab hauv lub receiver chamber. Tom qab kev sim thauj, peb pom tias TR ntawm PRE zoo sib xws hauv ob qho kev qhia, thiab kev thauj mus los no tsis txaus thiab nqus tau tsis zoo ('Papp < ​​1.0="" ×="" 10–6="" cm="" s)="" [3],="" thiab="" nws="" tsis="" tau="" qhia="" tias="" efflux,="" tawm="" tswv="" yim="" tias="" passive="" diffusion="" down="" ib="" concentration="" gradient="" yog="" lub="" ntsiab="" thauj="" mechanism="" ntawm="" pre="" (fig.="" 5).="" tsis="" tas="" li="" ntawd,="" cov="" yam="" ntxwv="" ntawm="" kev="" thauj="" mus="" los="" ntawm="" pre="" zoo="" ib="" yam="" nrog="" cov="" ac,="" is,="" thiab="" ec,="" cov="" khoom="" siv="" tau="" zoo="" uas="" ua="" rau="" muaj="" kev="" tiv="" thaiv="" oxidative="" hauv="" pre="" (fig.="" 4="" thiab="" table="" 4).="" cov="" txiaj="" ntsig="" no="" tau="" xav="" tau="" thiab="" qhia="" tau="" tias="" tam="" sim="" no="" bioassay="" system="" tsim="" nyog="" thiab="" txhim="" khu="" kev="" qha="" rau="" kev="" soj="" ntsuam="" ntawm="" cov="" yam="" ntxwv="" ntawm="" kev="" thauj="" mus="" los="" ntawm="" aig="" hauv="" pre="" hauv="" kev="" sib="" txawv="" caco-2="">

Hauv kev sib piv rau tus nqi canonical Papp, uas feem ntau siv los cuam tshuam txog kev thauj mus los ntawm ib qho kev sib xyaw, 'Papp tus nqi tau los ntawm TR raws li qhov kev cuam tshuam ntawm cov ntsiab lus nkhaus yuav tsis tsuas yog cuam tshuam cov khoom uas nkag mus rau monolayers nrog cov qauv tsis zoo tab sis Kuj tseem muaj lwm yam cuam tshuam nrog rau lub hom phiaj kev ua ub no, xws li cov khoom lag luam metabolites, cov khoom lag luam degraded, thiab txawm tias qee cov cytokines secreted los ntawm Caco-2 hlwb hauv teb rau stimulation uas tuaj yeem cuam tshuam rau lub hom phiaj bioactivity. Yog li, tus multifactor tau hais los saum toj no, tsis yog tsuas yog cov khoom thauj tsis zoo xwb, txiav txim siab 'Papp tus nqi. Yog li ntawd, 'Papp tuaj yeem cuam tshuam ntau dua hauv Vivo cov xwm txheej dua li tus nqi canonical Papp [26]. Hauv txoj kev tshawb no, peb tau qhia meej txog qhov tsis txaus ntseeg thiab tsis zoo absorbed ntawm PRE raws li nws 'Papp tus nqi, thiab qhov xwm txheej no yuav cuam tshuam rau qhov ntau npaum li cas thiab lub sijhawm kho mob ntev ntawm CD [27]. Txawm li cas los xij, cov kev tshawb pom no yuav muab kev qhia ntau ntxiv rau cov kws kho mob hauv daim ntawv thov CD thaum tsuas yog cov yam ntxwv ntawm AIG feem ntau hauv CD, tsis yog PhGs xwb, tau txheeb xyuas.

Txawm li cas los xij, cov khoom siv bioactivity uas tau xaiv yuav tsum muaj kev nkag siab txaus kom pom nyob rau hauv nruab nrab ntawm lub chaw txais khoom, uas nthuav tawm qhov kev sib tw rau kev tsim cov txheej txheem bioassay los ntsuas cov yam ntxwv ntawm kev thauj mus los ntawm PE. Tsis tas li ntawd, bioactivity yuav tsum nyob ntawm ntau yam khoom raws li qhov ua tau. Hauv peb txoj kev tshawb fawb, tag nrho cov tshuaj tiv thaiv oxidative muaj peev xwm tsim nyog tshaj li ob peb lwm txoj hauv kev (S1 Fig.). Tab sis txawm li ntawd los, peb qhov kev soj ntsuam tsuas tuaj yeem ntes TR ntawm PRE ntawm 120 min.

Sib sau ua ke, txoj kev tshawb fawb tam sim no tau tsim cov txheej txheem bioassay tshiab los ntsuam xyuas cov hnyuv permeability ntawm PRE nyob rau hauv sib txawv Caco-2 hlwb. Cov txiaj ntsig tau pom tau tias qhov tsis zoo absorbed passive diffusion down ib concentration gradient yam tsis muaj efflux yog lub ntsiab thauj mechanism ntawm PRE (Fig. 5), uas muab lub hauv paus pharmacokinetic rau kev kho mob daim ntawv thov ntawm PhGs nyob rau hauv CD. Kev siv cov txheej txheem bioassay tshiab los ntsuas TR thiab yog li xam cov 'Papp qhov tseem ceeb los ntsuas cov khoom siv hauv plab hnyuv ntawm AIG hauv PEs yog qhov ua tau kom meej meej. Txoj hauv kev no kuj tseem yuav tsim nyog rau lwm yam PE muab kev tsim nyog bioactivity

Cov ntaub ntawv txhawb nqa

S1 Fig. Cov teebmeem ntawm PRE ntawm (A) superoxide anion, (B) hydroxyl radical, (C) lipid peroxidation khoom, thiab (D) DPPH radical. Cov txheej txheem ntsuas tau ua raws li cov ntaub ntawv dhau los [1, 2] yam tsis tau siv cov khoom thauj. IC50 (50 feem pua ​​​​ inhibition concentration) thiab SC50 (50 feem pua ​​​​scavenging concentration) qhov tseem ceeb raug xam raws li tus qauv concentration- teb curves. (DOCX)

Sau Kev Pab Txhawb

Tsim thiab tsim cov kev sim: YG XC YQ. Ua qhov kev sim: YG CZ FL LF RC YS. Kev txheeb xyuas cov ntaub ntawv: CZ YG YQ. Pab txhawb cov tshuaj / cov khoom siv / cov cuab yeej tshuaj ntsuam: RC. Sau daim ntawv: YG YQ

Department of Research Center for Pharmacology and Toxicology, Institute of Medicinal Plant Development, Suav Academy of Medical Sciences & Peking Union Medical College, Beijing, PR Tuam Tshoj,

Heilongjiang University of suav tshuaj, Harbin, Heilongjiang, PR Tuam Tshoj