Kev Ntsuam Xyuas Ntawm Txoj hnyuv Thauj Khoom Ntawm Ib Phenylethanoid Glycoside-Rich Extract Los Ntawm Cistanche Deserticola Hla Lub Caco-2 Cell Monolayer Model

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 sib txawv ntawm cov hlwb paub tab thiab tsim cov monolayers tsis zoo. Cov hlwb uas nyob ib sab adhere ntawm nruj junctions tsim nyob rau hauv lub apical sab ntawm lub monolayer, uas muaj peev xwm cais cov passively.thiab nquag thauj cov tshuaj hla cov txheej txheem epithelial. 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.

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. 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 sijhawm, 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.

Cistanche deserticola YC Ma (CD), cov nroj tsuag holoparasitic, yog ib hom tshuaj suav tshuaj suav tshuaj feem ntau 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.Phenylethanoid glycosides (PhGs), suav nrog echinacoside (EC), acteoside (AC), thiab isoacteoside (IS), thiab lwm yam, yog ib chav kawm ntawm polyphenolic compounds. Lawv suav hais tias yog ib qho tseem ceeb ntawm cov khoom siv bioactive ntawm Cistanche hom. Cov kev tshawb fawb pharmacological tau pom tias bioactivity ntawm PhGs muaj ntau haiv neeg thiab suav nrog kev tiv thaiv oxidative, tiv thaiv qaug zog, hepatoprotective, immunomodulatory, anti-inflammatory, thiab neuroprotective teebmeem. Txawm li cas los xij, cov yam ntxwv ntawm txoj hnyuv thauj mus los ntawm PhGs tsis tau tshawb xyuas. 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.

Cistanche tubulosa extract

NATURAL CISTANCHE TUBULOSA rau kev txhim kho kev sib deev PHGS75% ECH 30% ACT 12%

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%) tau yuav los ntawm Yuav Tsum Biotechnology Co. (Chengdu, Suav). Dulbecco's modified Eagle's medium (DMEM), fetal bovine serum (FBS), thiab non-tseem ceeb 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 CD

Cov khoom siv cua qhuav CD yog hmoov thiab muab rho tawm los ntawm percolation nrog 70% ethanol. PhG-nplua nuj feem tau npaj raws li tau piav qhia yav dhau los thiab muab rho tawm nrog dej-saturated n-butyl cawv. Cov dej cawv extract tau concentrated thiab qhuav nyob rau hauv txo siab. Macroporous resin-UV spectrophotometric ntsuas cov ntsiab lus PhG ntawm 78.4%. Cov qauv kawg sawv cev rau 1.75% tawm los 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% 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 ntawm 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 haujlwm hauv qab no: kev ruaj ntseg, linearity, rhiab heev, precision (intra- thiab inter-hnub sib txawv), 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 heev nyob rau hauv lub xub ntiag ntawm vitamin C (0.4%) vim hais tias tus txheeb ze tus qauv sib txawv (RSD) qhov tseem ceeb nyob rau hauv lub ncov cheeb tsam yog <3.5%. 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% 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 pub dhau peb (siab, nruab nrab, thiab qis) concentration. Cov txiaj ntsig rov qab los ntawm cov txheej txheem rau cov tshuaj ntsuam xyuas tau muab sau tseg rau hauv Table 3. Qhov nruab nrab qhov rov qab los ntawm 90.0% mus rau 96.4%, qhia qhov tseeb zoo. Hauv kev xaus, cov txheej txheem HPLC tau tsim muaj kev txaus siab txog linearity, rhiab heev, precision, thiab raug rau qhov kom muaj nuj nqis ntawm AC, IS, thiab EC hauv kev thauj tsis.

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 qhov kev ntsuas nkhaus. Qhov ntau npaum li cas ntawm linearity yog 0. 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 tsom xam 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% thiab 4.72% rau intra- thiab inter-hnub sib txawv, raws li, qhia tau hais tias zoo precision. Yog li, txoj kev tsim bioassay yog txaus siab txog linearity, precision, thiab raug rau qhov kom muaj nuj nqis ntawm PRE nyob rau hauv ib tug thauj tsis.

Cell kab lis kev cai

Caco{{0}} hlwb raug coj los ntawm 37 degree thiab 95% txheeb ze av noo nyob rau hauv ib qho chaw uas muaj 5% CO2 thiab nruab nrab muaj DMEM, 10% FBS, 1% NEAA, 1% 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% 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.

cistanche tubulosa extract

NATURAL CISTANCHE TUBULOSA FOR SOOTHING THIAB LAXATIVE PHGS75% ECH 30% ACT 12%

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 rau basolateral (BL) cov kev tshawb fawb kev qhia lossis BL chamber (2.5 ml) hauv BL rau AP cov lus qhia. Rau AC, IS, thiab EC kev soj ntsuam 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% (w/v) vitamin C tau ntxiv rau kev ruaj khov cov qauv los txiav txim siab AC, IS, thiab EC cov ntsiab lus los ntawm HPLC, thiab cov qauv rau PRE bioassay yog assayed tam sim ntawd 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 raug 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 raug nthuav tawm raws li qhov txhais tau tias ± SD.

Cov txiaj ntsig

Kev sib xyaw ntawm AC, IS, thiab EC hauv PRE los ntawm CD

Vim tias AC, IS, thiab EC suav hais tias yog bioactive lojPhGs ntawm Cistanche hom, peb 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%, 1.84%, thiab 32.83%, feem. Yog li, peb lub tebchaw no suav nrog 61.27% ntawm PRE; Ntxiv mus, cov txiaj ntsig saum toj no qhia tau tias PhG cov ntsiab lus hauv PRE yog 78.4%. Yog li, AC, IS, thiab EC suav txog kwv yees li 80% ntawm PhGs hauv PRE.

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

Kev tshawb fawb ntawm covanti-oxidative kev ua haujlwm ntawm PhGslos ntawm CD tau tshaj tawm, thiab AC, IS, thiab EC account rau kwv yees li 80% 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 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 kwv yees li 60% ntawm PRE, AC thiab EC pom muaj zog tag nrho cov kev tiv thaiv oxidative ntau dua li PRE thiab IS. Vim tias IS (1.84%) yog ib feem me me ntawm PRE, lwm yam khoom siv, xws li cov tshuaj tiv thaiv tsis muaj zog tiv thaiv IS, kuj tseem ua kom pom tseeb hauv PRE. Kuj ceeb tias, tag nrho cov tshuaj tiv thaiv oxidative sib txawv ze li ntawm 7-fold ntawm AC thiab nws cov isomer IS, qhia tias tsis yog cov phenolic hydroxyl pab pawg nkaus xwb tab sis kuj yog txoj hauj lwm ntawm phenolic hydroxyl pawg hauv cov molecule cuam tshuam cov nyhuv anti-oxidative. .

Kev lees paub ntawm Caco-2 monolayers

Txhawm rau kom siv tau Caco-2 cell monolayer system, Papp qhov tseem ceeb ntawm propranolol (ib qho cim thauj tau zoo) thiab Lucifer daj (tus cim thauj tsis zoo) los ntawm AP mus rau BL hla Caco-2 monolayers tau txiav txim siab 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. Qhov kev ntsuam xyuas kev ua alkaline phosphatase kuj tau lees paub tias Caco-2 cell monolayers tau zoo piv rau cov hnyuv me me epithelium.

Lub permeability ntawm AC, IS, thiab EC

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 were nearly on the order of 10–7 cm/s, indicating that these compounds were poorly permeable. Furthermore, efflux or active transport was not observed because the ratios of Papp BL to AP / Papp AP to BL for AC, IS, and EC were between 0.90 ~ 1.55, and the criterion of net efflux proposed by the FDA Guidance is a ratio less than 2. 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 diffusion.

Lub permeability ntawm PRE

Cov txiaj ntsig saum toj no qhia tau tias tag nrho cov tshuaj tiv thaiv oxidative muaj peev xwm ntawm PRE tsawg kawg yog vim 61.27% 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 tshuaj tiv thaiv oxidative muaj peev xwm thiab suav cov 'Papp qhov tseem ceeb los cuam tshuam cov yam ntxwv ntawm kev thauj mus los 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. Ntxiv mus, qhov bidirectional TR ntawm PRE nce linearly ntawm kwv yees li 300 thiab 900 ug / ml (Fig. 5). Qhov tsis muaj kev xav tau ntawm qhov kev xav tau qhia tias qhov kev sib kis tsis zoo yog qhov tseem ceeb ntawm kev thauj mus los ntawm PRE.

cistanche tubulosa extract

NATURAL CISTANCHE TUBULOSA rau kev txhim kho kev tiv thaiv kab mob PHGS75% ECH 30% ACT 12%

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 cov kev soj ntsuam ntawm cov yam ntxwv thauj ntawm PE. Txhawm rau ntsuas cov khoom thauj ntawm PE ntau dua kev tshawb fawb, qee cov kws tshawb fawb tau txheeb xyuas ntau yam khoom, 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 siv hauv PE tso saib cov yam ntxwv sib txawv kiag li, 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 media 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 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 oxidative muaj peev xwm 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 zoo thiab nqus tau tsis zoo ('Papp < 1.0 × 10–6 cm / s), thiab nws tsis tau qhia efflux, qhia 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 cov kab sib txawv Caco{10}}.

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 haujlwm, xws li cov khoom lag luam metabolites, cov khoom lag luam degraded, thiab txawm tias qee cov cytokines secreted los ntawm Caco-2 hlwb teb rau stimulation uas yuav cuam tshuam rau lub hom phiaj bioactivity. Yog li, tus multifactor tau hais los saum 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 zog nrog hauv vivo cov xwm txheej dua li tus nqi canonical Papp. Hauv txoj kev tshawb no, peb tau qhia meej txog qhov tsis zoo thiab tsis zoo ntawm PRE raws li nws qhov 'Papp tus nqi, thiab qhov xwm txheej no yuav cuam tshuam nrog kev siv ntau npaum li cas thiab lub sijhawm kho mob ntev ntawm CD. 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 receiver chamber, uas ua rau muaj kev sib tw rau kev tsim lub tshuab bioassay los ntsuas cov yam ntxwv ntawm kev thauj mus los ntawm PE. Tsis tas li ntawd, bioactivity yuav tsum nyob ntawm ntau yam 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 hauv cov Caco-2 sib txawv. 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 plab hnyuv thauj khoom ntawm AIG hauv PEs yog ua tau. Txoj hauv kev no kuj tseem yuav tsim nyog rau lwm yam PE muab kev tsim nyog bioactivity.