mwwXaiv lus
Tsev / Kev paub / Ntsiab lus

Kev paub

Antioxidant Thiab Anti-Fatigue Constituents ntawm Okra

Mar 21, 2022

Fangbo Xia, Yu Zhong, Mengqiu Li, Qi Chang, Yonghong Liao, Xinmin Liu * thiab Ruile Pan *


Lub koom haum ntawm Kev Txhim Kho Kev Kho Mob, Peking Union Medical College, Suav Academy ntawm Kev Kho MobSciences, No 151, North Road Malianwa, Haidian District, Beijing 100193


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



Abstract


Okra (Abelmoschus esculentus (L.) Moench), cov zaub noj qab nyob zoo, tau dav dav hauv thaj chaw sov thiab thaj chaw sov. Cov kev tshawb fawb yav dhau los tau ua pov thawj tias okra pods muajtiv thaiv qaug zog,thiab lub hom phiaj ntawm qhov kev tshawb fawb no yog los qhia meej txog qhovtiv thaiv qaug zogcov pov thawj. Txhawm rau ua tiav qhov no, peb faib cov noob txiv ntoo (OPD) rau hauv cov noob (OSD) thiab tawv nqaij (OSK) thiab muab piv rau cov ntsiab lus ntawm tag nrho cov polysaccharides, tag nrho cov polyphenols, tag nrho flavonoids, isoquercitrin, thiab quercetin-3-O-gentiobiose thiab cov antioxidant. kev ua haujlwm hauv vitro thiabkev tiv thaiv kev qaug zoghauv vivo ntawm OSD thiab OSK. Cov ntsiab lus ntawm tag nrho cov polyphenols thiab tag nrho polysaccharides yog 29.5 feem pua ​​thiab 14.8 feem pua ​​hauv OSD thiab 1.25 feem pua ​​thiab 43.1 feem pua ​​hauv OSK, feem. Tag nrho cov flavonoids, isoquercitrin, thiab quercetin -3-O-gentiobiose (5.35 feem pua ​​, 2.067 feem pua ​​, thiab 2.741 feem pua ​​, raws li) tsuas yog kuaj pom hauv OSD. Antioxidant assays, suav nrog 1-diphenyl{17}}picrylhydrazyl (DPPH) scavenging, ferric txo antioxidant zog (FRAP) thiab txo hwj chim kuaj, thiab qhov hnyav-loaded ua luam dej kuaj pom OSD muaj cov antioxidant tseem ceeb thiablos tiv thaiv qaug zog. Ntxiv mus, kev txiav txim siab biochemical qhia tau hais tiaskev tiv thaiv kev qaug zogntawm OSD yog tshwm sim los ntawm kev txo cov ntshav lactic acid (BLA) thiab urea nitrogen (BUN), txhim kho hepatic glycogen cia thiab txhawb kev muaj peev xwm antioxidant los ntawm kev txo qis malondialdehyde (MDA) qib thiab nce superoxide dismutase (SOD) thiab glutathione peroxidase (GSH- Px) qib. Cov txiaj ntsig no tau ua pov thawj tias okra noob yog qhovtiv thaiv qaug zogib feem ntawm okra pods thiab polyphenols thiab flavonoids yog cov khoom xyaw nquag.


Ntsiab lus: okra; tiv thaiv qaug zog; antioxidant; polyphenols



Cistanche

1. Taw qhia


Kev qaug zog yog ib qho teeb meem ntawm lub cev, uas txhais tau hais tias nyuaj rau kev pib lossis txhawb nqa kev yeem ua haujlwm [1]. Sib nrug los ntawm kev muaj hnub nyoog zuj zus tuaj thiab nthuav tawm hauv kev mob qog noj ntshav, kev nyuaj siab, kab mob HIV, ntau yam sclerosis, thiab cov neeg mob Parkinson tus kab mob, kev qaug zog tau dhau los ua cov tsos mob tshwm sim ntau dua hauv cov tib neeg ib txwm muaj nrog kev nce qib ntawm lub neej niaj hnub [2-4]. Cov kev tshawb fawb hauv zej zog loj tau pom tias ntau dua ib nrab ntawm cov neeg laus yws yws txog kev qaug zog [5,6]. Ntev mus ntev los yog qaug zog ntev tsis tsuas yog txo qis lub neej zoo, tab sis kuj ua rau mob nkees nkees thiab lwm yam kab mob, thiab tseem ua rau karoshi (qhov txawv txav tuag los ntawm cov kab mob plawv los ntawm kev ua haujlwm ntau dhau [7]). Muaj ntau txoj kev xav txog qhov ua rau qaug zog, xws li kev qaug zog, uas hais tias kev qaug zog yog tshwm sim los ntawm qhov tsis muaj ATP rau actin-myosin coupling, Na` / K` siv thiab Ca2` uptake los ntawm sarcoplasmic reticulum [8-11. ]; homeostasis cuam tshuam txoj kev xav, uas qhia tias qaug zog yog tshwm sim los ntawm tsub zuj zuj ntawm ntau yam metabolic byproducts [10,12]; thiab kev puas tsuaj txoj kev xav qhia tias kev qaug zog tuaj yeem tshwm sim vim tsis ua haujlwm ntawm ib lossis ntau qhov chaw raws txoj hauv kev ntawm kev tsim hluav taws xob [13]. Ntawm cov no, oxidative kev nyuaj siab yog dav xav tias ua lub luag haujlwm tseem ceeb hauv etiology ntawm qaug zog [10,13–16]. Antioxidants los ntawm qee cov tshuaj ntsuab tau pom tias muaj txiaj ntsig los tiv thaiv kev qaug zog, uas qhia tau hais tias kev kho antioxidant yuav yog qhov muaj txiaj ntsig zoo los tiv thaiv kev qaug zog [17-20]. Yog li ntawd, cov neeg siv khoom tab tom nrhiav ntau cov tshuaj tiv thaiv antioxidant hauv lawv cov khoom noj kom txo tau oxidative puas tsuaj thiab tawm tsam qaug zog. Okra (Abelmoschus esculentus (L.) Moench, Tsev Neeg: Malvaceae), tseem hu ua poj niam ntiv tes, bhindi, thiab gumbo, yog cov nroj tsuag txhua xyoo nyob rau hauv teb chaws Africa thiab tau loj hlob nyob rau hauv ntau lub teb chaws thoob ntiaj teb, feem ntau nyob rau hauv tropical, subtropical, thiab thaj chaw sov sov.


Cov pods ntawm okra tau ntev tau siv los ua zaub thiab ib qhov chaw ntawm cov tshuaj noj. Hauv cov tshuaj Asian thiab African ib txwm siv, okra pods yog siv los ua cov khoom noj mucilaginous los tawm tsam gastritis [21]. Cov kev tshawb fawb pharmacological tau qhia tias okra muaj cov tshuaj tiv thaiv kab mob antioxidant, neuroprotective, antidiabetic, antihyperlipidemic, thiab tiv thaiv kev qaug zog [22-26]. Txawm hais tias kev ua haujlwm los tiv thaiv kev qaug zog ntawm okra pods tau raug pov thawj ua ntej [26], me ntsis paub txog nws cov haujlwm tiv thaiv kev qaug zog thiab cov khoom siv. Cov ntaub ntawv yav dhau los tau tshaj tawm tias cov txiv ntoo okra muaj cov ntsiab lus siab ntawm polysaccharides, polyphenols, thiab flavonoids [27–34]. Nws kuj tau pom tias polyphenols thiab flavonoids muaj zog antioxidant thiab tiv thaiv kev qaug zog hauv kev tshawb fawb yav dhau los [15,18,20,35,36]. Dab tsi ntxiv, peb qhov kev sim ua ntej ua tau zoo, uas peb siv cov tshuaj chloride ferric los ua cov xim tsim reagent, qhia tias okra noob extract pom muaj zog phenolic cov tshuaj tiv thaiv; Txawm li cas los xij, nws cov tawv nqaij extract tsis pom zoo li cov tshuaj tiv thaiv (cov ntaub ntawv ntxiv), uas pom tau tias cov noob txiv ntoo muaj cov ntsiab lus siab dua ntawm polyphenols thiab flavonoids dua li cov tawv nqaij okra. Yog li, nws tuaj yeem kwv yees tias cov noob txiv ntoo tuaj yeem yog cov tshuaj tiv thaiv kev qaug zog ntawm cov txiv hmab txiv ntoo, thiab polyphenols thiab flavonoids yuav yog cov khoom xyaw nquag. Lub hom phiaj ntawm txoj kev tshawb no yog los qhia meej txog cov tshuaj tiv thaiv kev qaug zog ntawm okra pods. Txhawm rau ua tiav qhov no, peb faib cov txiv hmab txiv ntoo tshiab rau hauv cov noob thiab tawv nqaij thiab muab piv rau cov ntsiab lus tshuaj ntawm tag nrho cov polysaccharides, tag nrho cov polyphenols, tag nrho flavonoids, isoquercitrin, thiab quercetin-3-O-gentiobiose, thiab cov haujlwm antioxidant hauv vitro thiab anti- qaug zog ua si hauv vivo ntawm okra noob thiab tawv nqaij. Tag nrho cov txiaj ntsig no tau qhia tias cov noob okra yog cov tshuaj tiv thaiv kev qaug zog ntawm okra pods, thiab polyphenols thiab flavonoids yog cov koom haum nquag. Lub peev xwm mechanisms ntawm kev tiv thaiv kev qaug zog hauv okra kuj tau kawm.


2. Ntu kev sim


2.1. Cog Khoom thiab Extraction


Cov txiv hmab txiv ntoo tshiab tau yuav los ntawm kev ua lag luam (Sanya, Hainan Province, Tuam Tshoj) thaum Lub Xya Hli 2013. Cov nroj tsuag tau lees paub los ntawm xibfwb Bengang Zhang, Lub Tsev Kawm Ntawv ntawm Tshuaj Kho Mob, Suav Academy ntawm Kev Kho Mob Sciences, thiab Peking Union Medical College, Beijing, Tuam Tshoj, qhov twg daim ntawv pov thawj (No. 20130705) tau muab tso rau hauv lub koom haum Herbarium. Ob feem ntawm 2.5 Kg tshiab okra pods tau npaj. Ib feem tau lyophilized ncaj qha kom tau txais cov noob qoob loo qhuav (251.3 g). Lwm feem tau muab faib ua cov noob txiv ntoo thiab cov tawv nqaij tawv tawv, lyophilized kom tau, raws li, cov noob txiv ntoo qhuav (50.1 g) thiab qhuav okra tawv (200.5 g). Lub okra pods, noob, thiab tawv nqaij tau zoo hauv av thiab muab rho tawm nrog 1500 mL boiling dej txhua 1h (3 zaug). Txhua cov kua lim tau sib xyaw ua ke thiab ua kom muaj zog hauv lub tshuab nqus tsev, kom tawm cov khoom seem ntawm okra pods (OPD, 105.4 g), okra noob (OSD, 20.5 g), thiab okra tawv (OSK, 84.1 g), feem. Yog li, qhov piv ntawm cov extract ntawm OPD: OSK: OSD yog hais txog 5: 4: 1. Tag nrho cov qauv tau muab khaws cia rau ntawm '20 ˝C kom txog thaum tom ntej tshuaj ntsuam xyuas thiab kev sim tsiaj.


Acteoside of Cistanche

2.2. Tshuaj thiab Reagents


Isoquercitrin thiab quercetin-3-O-gentiobiose raug cais tawm ntawm okra pods los ntawm peb lub chaw soj nstuam, ntawm purities yog tshaj 98 feem pua, los ntawm high-performance kua chromatography (HPLC) tsom xam. Rutin, Gallic acid, thiab D-glucose qauv tau yuav los ntawm National Institute for Food and Drug Control (Beijing, Suav). Folin-Ciocalteu's phenol reagent (1N, Coolaber Science & Technology, Beijing, Suav) tau siv rau kev txiav txim siab ntawm phenols. Cov khoom siv Ferric txo qis antioxidant zog (FRAP) tau yuav los ntawm Beyotime Biotechnology (Beijing, Suav). Lub 1, 1-diphenyl-2-picrylhydrazyl (DPPH) thiab Trolox tau yuav los ntawm Sigma-Aldrich Inc. (St. Louis, MO, USA). Phosphate buffer (PBS, Gibco) tau yuav los ntawm Thermo Fisher (Waltham, MA, USA). Cov kuab tshuaj, suav nrog acetonitrile (Thermo Fisher, MA, USA) thiab formic acid (CNW, Dusseldorf, Lub teb chaws Yelemees), yog qib HPLC, thiab dej tau txais los ntawm Milli-Q dej purification system (Millipore, MA, USA). Cov khoom siv tshuaj ntsuam xyuas cov ntshav lactate (BLA), ntshav urea nitrogen (BUN), hepatic glycogen (HG), glutathione peroxidase (GSH-Px), thiobarbituric acid (TBA), thiab superoxide dismutase (SOD) tau yuav los ntawm Nanjing Jiancheng Biotechnology Lub koom haum (Nanjing, Suav). Tag nrho lwm cov tshuaj reagents thiab tshuaj siv nyob rau hauv txoj kev tshawb no yog ntawm analytical qib.


2.3. Tsiaj


Txiv neej ICR nas (20–22 g) tau yuav los ntawm Vital River Laboratories (Qualified No.: SCXK 2012-0001, Beijing, Suav). Cov nas tau nyob hauv pawg ntawm 6 tus tsiaj ib lub tawb nyob rau hauv qhov kub thiab txias (23 ˝C ˘ 2 ˝C) thiab av noo (50 feem pua ​​˘ 10 feem pua) ntawm 12/12-h lub voj voog / tsaus nti (lub teeb rau ntawm 8:00 teev sawv ntxov txog 8:00 teev tsaus ntuj). Cov tsiaj muaj kev nkag tau dawb rau kev noj zaub mov zoo thiab ua kom tsis muaj dej haus hauv SPF (Specific pathogen Free) tsiaj tsev. Tag nrho cov txheej txheem sim tau ua raws li kev saib xyuas thiab kev pom zoo los ntawm Academy of Experimental Animal Center ntawm lub koom haum ntawm Kev Txhim Kho Kev Kho Mob thiab ua raws li NIH Phau Ntawv Qhia rau Kev Saib Xyuas thiab Kev Siv Cov Tsiaj Looj (Eighth edition)[37].



2.4. Chemical Analysis ntawm OSD thiab OSK


2.4.1. Kev txiav txim siab ntawm Tag Nrho Flavonoid (TF) Cov Ntsiab Lus


Cov ntsiab lus TF tau ntsuas los ntawm NaNO{{0}}Al(NO)3 colorimetric assay raws li Cov Qauv Txheej Txheem Kev Ntsuas thiab Kev Ntsuas Cov Khoom Noj Ua Haujlwm, uas tau tshaj tawm los ntawm National Health thiab Family Planning Commission ntawm PRC hauv 2{{20}}03, nrog kev hloov kho me me. Luv luv, 50 mg qauv tau yaj nyob rau hauv 40 mL distilled dej thiab cov qauv tov tau muab rho tawm los ntawm 50 mL chloroform peb zaug. Tom qab ntawd, cov kua dej tau diluted rau 50 mL thiab 2 mL ntawm cov tshuaj diluted yog adsorbed los ntawm 5 g polyamide resin thiab eluted nrog 20 mL 70 feem pua ​​​​ethanol. Lub 70 feem pua ​​​​ethanol elution yog concentrated rau 10 mL nyob rau hauv lub tshuab nqus tsev vacuum. Peb milliliters ntawm concentrated elution tau muab tso rau hauv 10 mL volumetric hwj, thiab ces 2 mL 70 feem pua ​​​​ethanol, 0.3 mL NaNO2 (5 feem pua), thiab 0.3 mL Al (NO) 3 ntxiv. Tom qab 5 feeb, 2 mL NaOH (1.0 M) tov tau ntxiv rau cov tshuaj tov thiab tso cai sawv ntsug rau 6 min. Thaum kawg, 2.4 mL 30 feem pua ​​​​ethanol tau ntxiv. Qhov nqus tau raug ntsuas tawm tsam qhov khoob ntawm 510 nm nrog UNICO -2012 UV-VIS spectrophotometer (Shanghai, Suav). Tag nrho cov ntsiab lus flavonoid tau suav los ntawm kev ntsuas nkhaus siv rutin raws li tus qauv (20-100 mg / L). Cov txiaj ntsig tau tshaj tawm tias yog mg ntawm rutin sib npaug ib mg ntawm cov qauv.


Echinacoside of Cistanche

2.4.2. Kev txiav txim siab ntawm Tag Nrho Polyphenol (TP) Cov Ntsiab Lus


Cov ntsiab lus TP tau txiav txim siab siv Folin-the Ciocaiteu txoj kev nrog kev hloov kho me ntsis [38]. Hauv luv luv, 50 mg qauv yog tov nrog 25 mL ntawm 50 feem pua ​​methanol tov, ces 0.5 mL qauv tov, 0.3 mL Folin-Ciocaiteu's reagent thiab 10 mL sodium carbonate (10 feem pua) tau txaus. sib tov, thiab tom qab ntawd lub ntim tau hloov mus rau 25 mL nrog dej distilled. Cov sib tov tau tso cai sawv ntsug ntawm 50 ˝C hauv qhov tsaus ntuj rau 1 teev. Absorbance tau ntsuas ntawm 765 nm. Ib qho calibration nkhaus ntawm Gallic acid tau npaj. Cov txiaj ntsig tau qhia tias yog mg ntawm Gallic acid sib npaug ib mg ntawm cov qauv.


2.4.3. Kev txiav txim siab ntawm Isoquercitrin thiab Quercetin -3-O-Gentiobiose


Quantification ntawm isoquercitrin thiab quercetin{{0}}O-gentiobiose tau ua los ntawm HPLC-UV siv tsib-point calibration nkhaus (r2=0.999) nyob rau hauv thaj tsam ntawm 5–5{{23 }}0 µg/mL. Isoquercitrin thiab quercetin-3-O-gentiobiose (5 mg, feem) thiab cov qauv (10 mg) tau yaj hauv 10 mL methanol, feem. Tag nrho cov kev daws teeb meem raug lim los ntawm 0.45-µm polytetrafluoroethylene fifilters ua ntej HPLC tsom xam. Waters 600 HPLC twj tso kua mis nrog lub Waters 2489 UV / Visible detector tau ua hauj lwm los cais cov tebchaw los ntawm ib tug reversed-phase kem (Thermo synchronize aQ, 4.6 mm ˆ 250 mm, 5 µm) ntawm tus khub tus nqi ntawm 1 mL / min. Cov theem txawb muaj xws li acetonitrile (A) thiab 0.1 feem pua ​​​​acetic acid hauv dej (B). Qhov kev pab cuam gradient elution yog raws li nram no: 10 feem pua ​​A ntawm 0 min thiab linearly mus txog 40 feem pua ​​ntawm 40 min, 90 feem pua ​​ntawm 41 mus rau 50 min, thiab 10 feem pua ​​ntawm 51–65 min. Lub wavelength rau UV nrhiav tau yog 354 nm thiab cov kab ke kub tau teem rau ntawm 25 ˝C. Cov tebchaw tau txheeb xyuas los ntawm kev sib piv nrog lub sijhawm khaws cia ntawm cov qauv thiab ntsuas los ntawm kev suav cov cheeb tsam hauv qab qhov nkhaus nrog cov qauv sab nraud.


2.4.4. Kev txiav txim siab ntawm Tag Nrho Polysaccharide (TPS) cov ntsiab lus


Cov ntsiab lus TPS tau txiav txim siab nrog txoj kev phenol-sulfuric acid [39,40]. Hauv luv luv, 80 mg qauv tau yaj hauv 50 mL dej distilled, thiab tom qab ntawd cov protein raug tshem tawm nrog txoj kev lim hiam (cov qauv tshuaj: chloroform: N-butyl cawv=20}: 4: 1). Tom qab ntawd, ib qho plaub-fold ntawm ethanol tau ntxiv rau cov kua nplaum uas ua rau ua rau cov kua roj ntsha polysaccharide extract. Cov tshuaj ethanol tau centrifuged ntawm 617.4 ˆg rau 30 min kom tau cov dej nag. Lub precipitate tau ntxuav ua ntu zus nrog ethanol, acetone, thiab ether, thiab yaj hauv 250 mL dej distilled. Tom qab ntawd, 0.1 mL polysaccharide tov yog tov nrog 0.9 mL distilled dej, 1 mL 5 feem pua ​​ntawm phenol tov, thiab 5 mL sulfuric acid nyob rau hauv ib lub raj kuaj, thiab cia nyob rau hauv chav tsev kub rau 30 min. Lub absorbance tau ntsuas ntawm 490 nm nrog UNICO -2012 UV-VIS spectrophotometer (Shanghai, Suav). Ib qho calibration nkhaus ntawm qabzib tau tsim, thiab TPS cov ntsiab lus tau txiav txim siab los ntawm qhov sib npaug ntawm qhov sib npaug ntawm qhov nkhaus. Cov txiaj ntsig tau qhia tias yog mg ntawm cov piam thaj sib npaug ib mg ntawm cov qauv.


2.5. Hauv Vitro Antioxidant Assay


Lub peev xwm antioxidant ntawm OPD, OSK, thiab OSD tau kuaj pom los ntawm peb txoj hauv kev, suav nrog {{{0}}}diphenyl-2-picrylhydrazyl (DPPH) scavenging, ferric txo antioxidant zog (FRAP), thiab txo lub zog, raws li cov ntaub ntawv [19,41] nrog kev hloov kho me ntsis. Trolox tau siv los ua kev tswj hwm zoo, thiab cov txiaj ntsig tau qhia tias Trolox sib npaug muaj peev xwm antioxidant. Raws li OPD tau suav nrog OSK thiab OSD, txhawm rau teeb pom kev ua haujlwm ntawm cov khoom siv ntawm okra pods, qhov concentration ntawm OSK thiab OSD tau siv los ua OPD sib npaug. Raws li cov kev xeem ua ntej, qhov concentration ntau ntawm OPD, ODS, OSK yog 0.0{{10}}4–{14}}.8, {{17 }}. -{{40}}.16 mg/mL rau DPPH assay; 1. 0–4. thiab 0.1–4, 0.08–3.2 thiab 0.02–0.8 mg/mL rau kev txo hwj chim tshuaj ntsuam xyuas, ntsig txog rau DPPH assay, qauv tshuaj (50 µL) nrog ib tug proportional range (0.004–0.8 mg OPD/mL, 0.0032–0.64 mg OSK/ mL thiab 0.0008–0.16 mg OSD/mL) yog tov nrog DPPH tov (100 µL, 1.28 ˆ 10´4 mol/L) rau kev ntsuas cov dawb radical-scavenging kev ua (A1) thiab 95 feem pua ​​ethanol (100 µL) rau kev tswj. (A2). Cov dej distilled (50 µL) yog tov nrog DPPH tov (100 µL) rau qhov dawb paug (A0). Qhov nqus tau raug ntsuas ntawm 517 nm tom qab cov kev daws teeb meem sib xyaw thiab khaws cia hauv chav sov li 30 min. Lub peev xwm txhawm rau txhawm rau DPPH radical tau suav nrog kev sib npaug hauv qab no:


Flavonoids of Cistanche

Kev ua ub ua no (%){{0}}[ 1-(A1-A2/{A0]* 100 feem pua ​​. (1)


FRAP ntawm peb cov qauv kev daws teeb meem (1.0-4.0 mg OPD/mL, 0.8–3.2 mg OSK/mL, thiab 0.2– {{10}}.8 mg OSD/mL) tau kuaj pom raws li kev qhia ntawm Beyotime Institute of Biotechnology. Cov tshuaj diluted qauv (5 µL) yog tov nrog FRAP ua hauj lwm tov (18{22}} µL) thiab khaws cia rau 5 feeb ntawm 37 ˝C. Tom qab ntawd qhov absorbance ntawm cov tshuaj tiv thaiv sib tov tau sau tseg ntawm 593 nm. Tus qauv nkhaus tau npaj siv FeSO4, xws li ntawm 0.15 txog 1.5 hli. Hauv kev txiav txim siab txo hwj chim, cov qauv kev daws teeb meem (0.5 mL rau 0.1–4 mg OPD/mL, 0.08–3.2 mg OSK/mL thiab 0. , pH 6.6) thiab poov tshuaj ferricyanide tov (1 feem pua, w/v), thiab incubation yog nqa tawm rau 20 min ntawm 50 ˝C. Trichloroacetic acid (0.5 mL, 10 feem pua, w / v) tau ntxiv rau qhov sib tov, uas yog centrifuged ntawm 201.6 ˆ g rau 10 min. Tom qab ntawd, txheej txheej sab saud ntawm cov tshuaj (0.5 mL) yog diluted nrog dej distilled (0.5 mL) thiab ces ferric chloride (0.1 mL, 0.1 feem pua, w / v) ntxiv rau nws. Lub absorbance yog ntsuas ntawm 700 nm.



2.6. Los tiv thaiv kev qaug zog ntawm OPD, OSK thiab OSD thiab Biochemical Analysis


Raws li tib lub laj thawj raws li kev ntsuas hauv vitro antioxidant, cov koob tshuaj tiv thaiv kev qaug zog ntawm OPD, OSK thiab OSD tau tsim los ua OPD pawg (0.75, 1.50, thiab 3.{{ 6}} g/kg lub cev hnyav), OSK pawg (0.60, 1.20, thiab 2.40 g/kg lub cev hnyav) thiab OSD pawg (0.15, 0.30 thiab 0.60 g / kg lub cev hnyav). Cov nas tau noj nrog cov zaub mov yooj yim hauv qhov chaw sim ua ntej qhov kev sim. Tom qab 3 hnub ntawm kev hloov kho, 120 nas tau muab faib ua kaum pawg: pawg tswj hwm (CG), 3 OPD, OSK, thiab OSD pawg txhua. Txhua pawg muaj 12 nas. Pab pawg tswj hwm (CG) tau kho nrog dej tsis muaj menyuam, thaum lwm pab pawg tau kho nrog cov khoom sib txuas. Cov dej / cov khoom rho tawm tau raug tswj hwm qhov ncauj (8:{24}} teev sawv ntxov) rau cov nas ib hnub ib zaug rau 21 hnub, thiab kev sim qhib qhov chaw thiab qhov hnyav-loaded ua luam dej (WLST) tau ua rau hnub 20 thiab 21, raws li. Tom qab WLST, cov nas raug tshem tawm ntawm lub pas dej, qhuav nrog cov ntaub so ntswg, thiab muab tso rau hauv lawv cov qub tawb. Cov neeg sim ntawm WLST thiab qhov kev sim qhib teb tau ua rau qhov muag tsis pom qhov kev sim thiab cov tsiaj txhu tau randomized ua ntej koom nrog kev sim. Txhawm rau nthuav tawm cov txiaj ntsig los tiv thaiv kev qaug zog ntawm OSD thaum lub sijhawm rov qab los, nas tau muab tshuaj loog nrog ether thiab cov ntshav tau sau los ntawm txhua pab pawg tom qab WLST 24 teev. Cov qauv ntshav tau txais los ntawm centrifugation (896 ˆ g, 15 min, 4 ˝C). Daim siab tau muab tshem tawm sai sai thiab ntxuav hauv dej khov-txias saline. Tag nrho cov qauv tau khov rau hauv cov kua nitrogen thiab khaws cia ntawm '80 ˝C rau kev soj ntsuam ntxiv.


2.6.1. Qhib-Field Test


Txhawm rau txheeb xyuas qhov cuam tshuam ntawm OPD, OSK, thiab OSD ntawm cov haujlwm locomotor, cov nas raug soj ntsuam tau siv lub tshuab qhib lub khoos phis tawj-pab tswj tau tsim los ntawm peb [42,43]. Cov cuab yeej muaj plaub lub tso tsheb hlau luam (30 cm inch, 40 cm nyob rau hauv qhov siab) nrog lub koob yees duab video tsau rau saum. Kev sim tau ua nyob rau hauv ib chav nyob ntsiag to, thiab cov cuab yeej tau teeb pom kev zoo los ntawm lub teeb ci ntawm 120 Lux ntawm lub qab nthab. Ib teev tom qab txhaj tshuaj, txhua tus nas tau muab tso rau ntawm qhov chaw ntawm lub tank hlau thiab tso cai rau kev tshawb nrhiav dawb rau 2 min thiab qhov kev ncua deb mus rau hauv 4 min, uas yog qhov ntsuas qhov ntsuas ntawm lub tshuab ua haujlwm, suav nrog software. Plaub nas tau sim ib txhij.


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2.6.2. Weight-Loaded Swimming Test (WLST)


WLST tau ua los ua kom qaug zog raws li cov txheej txheem tau piav qhia yav dhau los nrog kev hloov kho me ntsis [19,44,45]. Ib teev tom qab noj tshuaj, cov nas tau ntim nrog cov ntawv txhuas uas txuas rau tib txoj hauj lwm ntawm lawv cov tails thiab hnyav li ntawm 3 feem pua ​​ntawm lawv lub cev hnyav. Tom qab ntawd cov nas uas hnyav hnyav tau raug yuam kom ua luam dej hauv cov neeg tau txais cylindrical (inch 40 cm), uas tau ntim nrog dej (25 ˘ 1 ˝C) mus rau qhov tob ntawm 30 cm. Kev qaug zog tau raug cais raws li kev poob ntawm kev sib koom ua ke thiab tsis tuaj yeem nce mus rau saum npoo li ntawm 10 s, thiab lub sijhawm ua luam dej tau sau tseg tam sim ntawd. Cov dej hauv pas dej tau hloov tom qab txhua qhov kev sib tham.



2.6.3. Kev soj ntsuam ntawm Biochemical Parameters Muaj feem xyuam rau qaug zog


Serum tau npaj rau kev txiav txim siab ntawm cov ntshav urea nitrogen (BUN) thiab ntshav lactic acid (BLA). Lub siab tau npaj rau kev txiav txim siab ntawm malondialdehyde (MDA) sib npaug, tag nrho superoxide dismutase (SOD), glutathione peroxidase (GSH-Px), thiab hepatic glycogen (HG). Txhua qhov kev ntsuas biochemical tau txiav txim siab siv cov khoom siv hauv kev lag luam, thiab tag nrho cov txheej txheem ua raws li cov chaw tsim khoom cov lus qhia. Luv luv, cov ntsiab lus ntawm HG tau txiav txim siab siv anthrone reagent ntawm 620 nm, thaum BLA tau txiav txim siab siv nitrotetrazolium xiav chloride reagent ntawm 530 nm. BUN tau soj ntsuam los ntawm urease nrog glutamate dehydrogenase ua ke enzyme txoj kev ntawm 640 nm. GSH-PX kev ua haujlwm tau ntsuas hauv kev sib txuas los ntawm kev ntsuas qhov txo qis ntawm triphosphopyridine nucleotide (NADPH) ntawm 340 nm. Kev soj ntsuam rau SOD kev ua haujlwm tau ua raws li nws lub peev xwm los tiv thaiv oxidation los ntawm O2 ua los ntawm xanthine-xanthine oxidase system. Ib chav tsev ntawm SOD kev ua haujlwm tau txhais tias yog tus nqi uas txo qhov nqus ntawm 450 nm los ntawm 50 feem pua. Cov ntsiab lus ntawm MDA sib npaug tau txiav txim siab ntawm 532 nm los ntawm kev ua haujlwm nrog TBA los tsim cov khoom lag luam chromophoric ruaj khov.


2.7. Kev txheeb cais


Kev txiav txim tshuaj tshuaj tau ua nyob rau hauv triplicate thiab cov txiaj ntsig tau qhia raws li qhov txiaj ntsig. Cov tshuaj tua kab mob antioxidant tau ua tiav hauv triplicate thiab cov txiaj ntsig tau qhia tias yog qhov tseem ceeb ntawm tus qauv sib txawv. Cov ntaub ntawv coj tus cwj pwm thiab biochemical tau txheeb xyuas los ntawm SPSS statistical software (SPSS 19.0 Inc., Chicago, IL, USA). Ib txoj kev tsom xam ntawm qhov sib txawv (Ib-txoj kev ANOVA) nrog qhov tsawg kawg nkaus qhov sib txawv (LSD) tau siv rau kev sib piv ntawm pab pawg. P tus nqi tsawg dua 0.05 tau suav tias yog qhov tseem ceeb. Cov txiaj ntsig tau nthuav tawm raws li qhov txhais tau tias yog tus qauv yuam kev ntawm qhov nruab nrab (SEM).


3. Cov txiaj ntsig


3.1. Chemical Analysis ntawm OSK thiab OSD


Raws li pom nyob rau hauv Table 1, cov ntsiab lus ntawm TF thiab TP hauv OSD tau ntau dua li cov hauv OSK, lub sijhawm no, cov ntsiab lus ntawm TPS hauv OSD qis dua li hauv OSK. Cov tebchaw ntawm isoquercitrin thiab quercetin-3-O-gentiobiose tsuas yog kuaj pom hauv OSD siv HPLC (pom hauv daim duab 1).


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3.2. Hauv Vitro Antioxidant Kev Ua Haujlwm ntawm OPD, OSK, thiab OSD


Raws li tau pom nyob rau hauv Table 2, cov kev ua antioxidant ntawm OPD thiab OSD tau ua pov thawj los ntawm DPPH, FRAP, thiab txo cov kev ntsuas hluav taws xob, thiab OSD tau pom tias muaj kev ua haujlwm antioxidant zoo dua li OPD. Txawm li cas los xij, OSK tau pom qhov tsis muaj zog hauv kev txo qis kev ntsuas hluav taws xob thiab tsis muaj txiaj ntsig hauv DPPH thiab FRAP kev xeem. Yog li, OSD yog ib feem ntawm okra pods rau kev ua haujlwm antioxidant



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3.3. Cov teebmeem ntawm Locomotor Kev Ua Haujlwm ntawm OPD, OSK, thiab OSD


Locomotor kev ua ub no ntawm cov nas tau sim nyob rau 20 hnub tom qab tau kho nrog OSD. Raws li pom nyob rau hauv daim duab 2, cov koob tshuaj ntau ntawm OSD pawg (0.6 g/kg) txo qis tag nrho cov nas nyob hauv qhov chaw tshiab raws li piv nrog rau pawg tswj hwm (p=0. 04). Txawm li cas los xij, tsis muaj qhov cuam tshuam tseem ceeb rau lwm pab pawg ntawm tag nrho qhov kev ncua deb tau pom. Cov txiaj ntsig no tau pom tias okra pods thiab noob tsis muaj kev cuam tshuam rau hauv nruab nrab paj hlwb. Ntxiv mus, nws tuaj yeem txiav txim siab tias OSD tuaj yeem muaj cov nyhuv sedative me ntsis ntawm koob tshuaj siab (0.6 g / kg).



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3.4. Qhov cuam tshuam rau lub sijhawm ua luam dej ntawm OPD, OSK, thiab OSD (WLST)


Raws li pom hauv daim duab 3, tom qab ntxiv OPD, OSK, thiab OSD rau 21 hnub, OPD (3 g/kg) thiab OSD (0.3 thiab 0.6 g/kg) tuaj yeem kho. ua kom cov nas ua luam dej ntev ntev, uas ncav cuag 57.84 ˘ 12.37, 46.51 ˘ 5.82 thiab 70.05 ˘ 12.07 min (p < 0="" .05,="" p="">< 0.05="" thiab="" p="">< 0.="" hauv="" pawg="" tswj="" hwm="" (15.20="" ˘="" 1.49="" min).="" lub="" caij="" no,="" opd="" (0.75="" thiab="" 1.5="" g="" kg),="" osd="" (0.15="" g="" kg)="" thiab="" osk="" (0.6,="" 1.2="" thiab="" 2.4="" g="" kg)="" kev="" kho="" mob="" tsis="" pom="" qhov="" nce="" ntawm="" lub="" sijhawm="" ua="" luam="" dej="" hauv="" nas="" (="" 14.73="" ˘="" 1.28,="" 23.96="" ˘="" 3.70="" thiab="" 27.34="" ˘="" 5.72="" min,="" ntsig="" txog)="" piv="" rau="" pawg="" tswj="" hwm="" (p="0.897," p="0.398" thiab="" p="0.268," ntsig="" txog="" ),="" tab="" sis="" osd="" (0.15="" g="" kg)="" tau="" pom="" qhov="" sib="" txawv="" ntawm="" lub="" sijhawm="" ua="" luam="" dej,="" tab="" sis="" tsis="" qhia="" qhov="" sib="" txawv="" tseem="" ceeb="" (p="0.225)." ntxiv="" mus,="" one-way="" anovas="" qhia="" qhov="" sib="" txawv="" tseem="" ceeb="" ntawm="" lub="" sijhawm="" ua="" luam="" dej="" ntawm="" cov="" nas="" ntawm="" pawg="" (f="" (3,="" 40)="12.839," p="">< 0.001).="" lub="" sijhawm="" ua="" luam="" dej="" ntawm="" cov="" koob="" tshuaj="" osd="" pawg="" siab="" (0.6="" g="" kg)="" tau="" pom="" qhov="" sib="" txawv="" raws="" li="" piv="" rau="" cov="" pab="" pawg="" nruab="" nrab="" koob="" (0.3="" g="" kg)="" (p="0.019)," thiab="" tib="" qhov="" txiaj="" ntsig="" tau="" pom="" nruab="" nrab="" ntawm="">

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3.5. Cov nyhuv ntawm OSD ntawm Biochemical Parameters nyob rau hauv nas tom qab qhov hnyav-loaded Swimming Test


Raws li qhov tshwm sim ntawm qhov ntsuas qhov hnyav-loaded ua luam dej pom, OSD tuaj yeem ua rau cov nas ua luam dej ntev ntev; Txawm li cas los xij, OSK kev kho mob tsis tau pom tias muaj kev nce ntxiv hauv lub sijhawm ua luam dej hauv nas. Yog li, biochemical tsis tau tsuas yog txiav txim siab ntawm OSD cov nas.


3.5.1. Qhov cuam tshuam ntawm OSD ntawm BLA, BUN, thiab HG


Qhov txawv ntawm BLA, BUN thiab HG cov ntsiab lus ntawm cov pab pawg yog qhov tseem ceeb (BLA: F (3, 40)=8.257, p < {{10}}.{{="" 20}}01;="" bun:="" f="" (3,="" 40)="8.596," p="">< 0.{{3{{32}="" }}}}01;="" hg:="" f="" (3,="" 36)="6.960," p="">< 0="" {40}}="" 01).="" raws="" li="" pom="" nyob="" rau="" hauv="" table="" 3,="" cov="" theem="" ntawm="" bla="" thiab="" bun="" hauv="" osd="" pawg="" tau="" qis="" dua="" qhov="" tseem="" ceeb="" hauv="" pawg="" tswj="" 24="" teev="" tom="" qab="" qhov="" hnyav="" loaded="" ua="" luam="" dej="" (bla:="" p="">< 0.05,="" p="">< 0.05="" thiab="" p=""><0.01 rau="" 0.15,="" 0.3="" thiab="" 0.6="" g="" kg,="" raws="" li;="" bun:="" p="">< 0.01="" rau="" 0.15,="" 0.3="" thiab="" 0.6="" g="" kg).="" cov="" theem="" ntawm="" hg="" hauv="" osd="" pawg="" tau="" siab="" dua="" li="" piv="" rau="" pawg="" tswj="" hwm="" (p="">< 0.05,="" p="">< 0.01="" thiab="" p="">< 0.01="" rau="" 0.15,="" 0.3="" thiab="" 0.6="" g="" kg,="" ntsig="">


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3.5.2. Cov teebmeem ntawm OSD ntawm GSH-PX, MDA, thiab SOD


Qhov sib txawv ntawm SOD, GSH-PX cov dej num thiab MDA sib npaug ntawm cov pab pawg yog qhov tseem ceeb (MDA: F (3, 40)=8.355, p < 0.{{18}="" }01;="" sod:="" f="" (3,="" 42)="9.876," p="">< 0.001;="" gsh-px:="" f="" (="" 3,="" 40)="7.959," p="">< 0="" 001).="" raws="" li="" pom="" nyob="" rau="" hauv="" table="" 4,="" qhov="" sib="" npaug="" ntawm="" mda="" hauv="" osd="" pawg="" tau="" qis="" dua="" li="" pawg="" tswj="" hwm="" tom="" qab="" qhov="" hnyav="" loaded="" ua="" luam="" dej="" (p="">< 0.01,="" p="">< 0.="" {45}}1="" thiab="" p="">< 0.="" tsis="" tas="" li="" ntawd,="" cov="" theem="" ntawm="" sod="" thiab="" gsh-px="" hauv="" osd="" pawg="" tau="" nce="" siab="" dua="" li="" piv="" rau="" pawg="" tswj="" hwm="" (sod:="" p="">< 0.01,="" p="">< 0.01="" thiab="" p="">< 0.01;="" gsh-px:="" p="">< 0.05,="" p="">< 0.01="" thiab="" p="">< 0.01="" rau="" 0.15,="" 0.3="" thiab="" 0.6="" g="" kg,="" ntsig="">


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4. Kev sib tham


Cov txiv hmab txiv ntoo tsis paub qab hau yog cov zaub noj qab haus huv uas tau noj hauv ntau thaj chaw ntawm lub ntiaj teb. Cov kev tshawb fawb yav dhau los tau tshaj tawm tias cov txiv hmab txiv ntoo tsis paub qab hau muaj cov tshuaj tua kab mob antioxidant thiab tiv thaiv kev qaug zog [25,46], tab sis nws cov khoom xyaw nquag thiab cov txheej txheem muaj peev xwm tsis meej. Txhawm rau tshawb xyuas cov tshuaj tiv thaiv antioxidant thiab tiv thaiv kev qaug zog ntawm okra pods, peb tau faib cov txiv hmab txiv ntoo tshiab rau hauv cov noob thiab tawv nqaij thiab tau txais OSD, OSK, thiab OPD los ntawm cov txheej txheem rho tawm, tom qab ntawd cov tshuaj antioxidant hauv vitro ntawm OSD, OSK, thiab OPD tau kuaj pom nrog DPPH, FRAP thiab txo lub zog thiab tiv thaiv kev qaug zog hauv vivo ntawm OSD, OSK thiab OPD tau kawm siv qhov hnyav-loaded ua luam dej. Peb cov txiaj ntsig tau pom tias OSD tsis tsuas yog muaj cov tshuaj tiv thaiv antioxidant zoo hauv vitro tab sis kuj tseem ua rau lub sijhawm ua luam dej ntawm cov nas tau ntev dua li piv rau pawg tswj hwm, thaum, OSK tsis pom qhov cuam tshuam rau ob qho tib si. Lub sij hawm tag nrho ntawm kev ua luam dej yog qhov ntsuas ncaj qha cuam tshuam txog kev ua haujlwm ntawm lub cev [47], thiab kev txhawb nqa ntawm kev ua kom muaj zog muaj feem cuam tshuam nrog lub sijhawm ua luam dej ntev dua. Yog li ntawd, cov tshuaj tiv thaiv antioxidant thiab tiv thaiv kev qaug zog ntawm OPD yuav tsum raug txiav tawm raws li nws cov noob. Ntxiv mus, hauv txoj kev tshawb no, OSD (0.3{{10}}}, 0.60 g/kg) tuaj yeem ua rau ntev nas ua luam dej, thiab tib neeg cov koob tshuaj ntawm cov noob okra yuav yog 0.9. ~ 1.8 g / kg (4.5 ~ 8.9 g / kg rau okra pods) raws li lub cev nto [48]. Cov koob tshuaj no tuaj yeem ncav cuag los ntawm tib neeg kev noj zaub mov. Txawm li cas los xij, cov khoom xyaw nquag ntawm okra tseem tsis meej. Cov txiaj ntsig ntawm kev tshuaj xyuas tshuaj hauv txoj kev tshawb fawb tam sim no tau pom tias cov ntsiab lus ntawm polyphenols hauv OSD yog kwv yees li 24 npaug ntau npaum li hauv OSK, tab sis cov ntsiab lus ntawm polysaccharides hauv OSD yog qis dua li hauv OSK (Table 1).


Moreover, total flavonoids content and two compounds of isoquercitrin and quercetin-3-O-gentiobiose were only detected in OSD and were not detected in OSK (Table 1). It has been proven that okra seeds contained epigallocatechin oligomers, catechin, and its oligomers, isoquercitrin, quercetin-3-O-gentiobiose, and other catechin and quercetin derivatives [27–29]. Furthermore, previous studies have reported that polyphenols like catechin and flavonoids like quercetin possess anti-fatigue activity due to their antioxidant activity [35,36,49–51]. Therefore, we deduced that polyphenols and flavonoids of OSD might be the antioxidant and anti-fatigue constituents. As for the reason that the effect of OPD was less than the effect of OSD, it might be that the high content of mucilaginous polysaccharides can affect the assimilation of polyphenols and flavonoids in OPD, which needs to be researched in the future. When it comes to the anti-fatigue mechanism of OSD, an open-field test showed that OSD had no central nerve stimulation in mice, which proved that the anti-fatigue of OSD was not through central excitation. Apart from this, it was likely that OSD possessed a slight sedative effect in high doses (>0.6 g/kg). Cov ntaub ntawv xav tau ntau ntxiv los ua pov thawj qhov no vim tias qhov txo qis hauv kev txav mus los kuj tuaj yeem yog qhov tshwm sim ntawm kev txo qis los tshawb nrhiav lossis muaj peev xwm anxiogenic nyhuv. Txawm li cas los xij, hauv qhov kev tshawb fawb no, qhov kev sim qhib-thaj chaw tau siv los tsis suav nrog qhov muaj peev xwm uas OSD tau ua haujlwm los tiv thaiv kev qaug zog los ntawm kev ua kom muaj zog ntawm lub paj hlwb hauv nruab nrab, thiab ntau cov ntaub ntawv los ua kom pom cov nyhuv sedative ntawm OSD tsis tuaj. Tsis tas li ntawd, ntau hom kab mob reactive oxide (ROS) uas tsim thaum lub sij hawm ua haujlwm hnyav tau raug pov thawj tias yog qhov tseem ceeb ntawm lub cev qaug zog. ROS ntau dhau tuaj yeem cuam tshuam rau ntau yam txheej txheem metabolic thiab enzymatic los ntawm kev tawm tsam polyunsaturated fatty acids los tsim MDA thiab ua rau tsis ua haujlwm ntawm biofilms [13,52].


Vim li no, ROS tsis tsuas yog cuam tshuam rau cov kev ua haujlwm enzymatic thaum lub sijhawm siv hluav taws xob thiab cov txheej txheem excitation-contraction coupling ncaj qha tab sis kuj tseem ua rau cov khoom pov tseg xws li BLA thiab BUN. Ntxiv mus, qhov nce qib ntawm BLA yog ib qho tseem ceeb ua rau qaug zog vim yog txo qis pH hauv cov ntaub so ntswg, thiab cov ntsiab lus ntau dua ntawm BUN qhia tias muaj zog ua haujlwm qis. Yog li ntawd, nws tuaj yeem tsis tsuas yog txhim kho kev ua kom muaj zog, tab sis kuj txo lub cev qaug zog thiab txhawb kev rov qab los ntawm kev txhawb nqa kev ua haujlwm ntawm enzymatic antioxidant systems suav nrog SOD thiab GSH-PX, lossis ncaj qha ntxiv antioxidants. Hauv txoj kev tshawb fawb tam sim no, kev txiav txim siab biochemical uas tau ua 24 teev tom qab kuaj ua luam dej qhia tau hais tias OSD kev kho mob txo qis BLA thiab BUN hauv cov ntshav, MDA hauv siab, thiab nce qib HG, SOD, thiab GSH hauv cov ntshav. daim siab thaum lub sij hawm rov qab qaug zog, uas ua pov thawj tias OSD tuaj yeem txo lub cev qaug zog thiab txhawb kev rov qab los. Xav txog cov txiaj ntsig ntawm kev soj ntsuam antioxidant hauv vitro hauv peb txoj kev tshawb fawb, nws tuaj yeem txiav txim siab tias OSD ua si los tiv thaiv kev qaug zog tsis yog los ntawm kev tshem tawm cov dawb radicals ncaj qha tab sis kuj los ntawm kev txhawb nqa cov dej num ntawm cov enzymes antioxidant xws li SOD thiab GSH-PX.


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Nyem daim duab rau Cistanche Uk rau kev qaug zog

5. Cov lus xaus


Hauv kev xaus, txoj kev tshawb fawb tam sim no tau qhia tias OSD yog cov tshuaj tiv thaiv kev qaug zog ntawm okra, thiab polyphenols thiab flavonoids yuav yog cov koom haum nquag ntawm OSD vim lawv cov kev ua haujlwm antioxidant. Txawm li cas los xij, kev qaug zog yog qhov tshwm sim nyuaj, thiab qhov kev tshawb fawb tam sim no ntawm kev tiv thaiv kev qaug zog ntawm okra yog qhov ua ntej. Cov ntsiab lus ntxiv tseem yuav tsum tau teeb pom kev yav tom ntej.



Cov ntaub ntawv


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