Lub tswv yim tshiab rau kev tshuaj xyuas cov khoom nquag hauv Cistanche Tubulosa Raws li Spectrum-Effect Relationship Analysis Thiab Network Pharmacology
Feb 13, 2023
Abstract
Cistanche tubulosa(Schenk) R. Wight yog cov tshuaj ntsuab muaj txiaj ntsig hauv Suav teb. Txoj kev tshawb no tsom los tshawb txog cov peev txheej ntawm cov txheej txheemC. tubulosantawmkev ua haujlwm antioxidantsiv spectrum-npaum kev sib raug zoo thiab networkpharmacology thiab muaj peev xwm ntawm kev siv tshuaj ntsuab dregs. Nyob rau hauv no ua hauj lwm, txawv extracts ntawmC. tubulosa, suav nrog cov khoom siv tshuaj ntsuab, cov dej rho tawm, thiab cov khoom siv tshuaj ntsuab, tau raug tshuaj xyuas siv cov cuab yeej ua haujlwm siab ua kua chromatography (HPLC). Tsis tas li ntawd, cov haujlwm antioxidant tau kwv yees hauv vitro, suav nrog 2, 2-diphenyl-1-picrylhydrazyl; superoxide anion; thiab hydroxyl radical scavenging kev soj ntsuam. Cov kev sib raug zoo ntawm spectrum-zoo ntawm HPLC cov ntiv tes thiab cov peev xwm lom tau raug tshuaj xyuas los ntawm ib nrab qhov tsawg kawg nkaus squares regression, bivariate correlation tsom xam, thiab redundancy tsom xam. Tsis tas li ntawd, network pharmacology tau siv los kwv yees qhovmuaj peev xwm mechanismsntawmC. tubulosanyob rau hauv kev kho mob ntawmcov kab mob antioxidant. Raws li cov txiaj ntsig, kaum ib lub ncov siab tau sib koom los ntawm cov khoom sib txawv. Geniposidic acid, echinacoside, verbascoside, tubuloside A, thiab isoacteoside tau txheeb xyuas thiab muab piv nrog ntau hom kev sib txawv.C. tubulosa. Txoj kev tshawb fawb txog kev sib raug zoo spectrum qhia tau hais tias ncov A6 yuav yog qhov txiav txim siab tshaj plaws ntawm peb daim ntawv. Raws li lub network pharmacology, muaj 159 lub hom phiaj cov noob sib koom los ntawm cov khoom nquag thiab cov kab mob antioxidant. Cov hom phiaj cuam tshuam txog kev ua haujlwm antioxidant thiab cov kev cuam tshuam tau raug tham. Peb cov txiaj ntsig tau muab lub hauv paus theoretical rau kev rov ua dua cov tshuaj ntsuab residues thiab lub peev xwm mechanisms ntawm C. tubulosa hauv kev kho cov kab mob antioxidant.
Nyem qhov no kom tau txais cov ntaub ntawv ntxiv txog Cistanche Active Components
Nug kom paub meej ntxiv: wallence.suen@wecistanche.com 0015292862950
1. Taw qhia
Cistanche tubulosa(Schenk) R. Wight, ib qho ntawm cov tshuaj nquag siv hauv tsev neeg Cistanche, yog hu ua "Ginseng ntawm suab puam" rau nws cov txiaj ntsig kev noj qab haus huv ntau [1]. Kev tshawb nrhiav tshuaj niaj hnub no tau tshawb pom tias tsev neeg Cistanche muaj ntau yam tshuaj, xws liantioxidant, tshuaj tiv thaiv kab mob, hypoglycemic, antidepressant, kev txawj ntse txhim kho, thiabantimicrobial teebmeem [2]. Thawj chemical pawg hauvCistanche tshuaj ntsuaba yog phenylethanoid glycosides (PhGs) [1]. PhGs muab los ntawm Cistanche hom tuaj yeem txhawb nqa cov haujlwm ntawm superoxide dismutase (SOD) thiab glutathione peroxidase (GSH-Px) [3]. Echinacoside, ib tug tswv cuab ntawm PhG tsev neeg, thim rov qab zoohauv vivooxidative kev nyuaj siab tshwm sim los ntawm cov khoom noj muaj qabzib siab los ntawm kev txo qis ntawm nitrous oxide system (NOS) kev ua haujlwm thiab phosphor-eNOS qhia [4]. Lub cev pob txha ntawm PhG muaj phenyl ethyl cawv thiab glycosyl moieties. PhGs ntseeg tau tias muaj cov haujlwm tseem ceeb vim yog phenolic hydroxyl pawg hauv lawv cov qauv. Kev ua haujlwm antioxidant ntawm PhGs nce nrog cov phenolic hydroxyl pawg [5].
Reactive oxygen hom (ROS) muaj lub luag haujlwm tseem ceeb hauv ntau cov txheej txheem physiological thiab cov txheej txheem tiv thaiv tseem ceeb. Kev nquag raug rau ROS siab ntau tuaj yeem ua rau tsis muaj kev puas tsuaj rau cov protein, lipids, thiab nucleic acids. ROSs tuaj yeem ua cov molecules nruab nrab (xws li hydrogen peroxide), ions (piv txwv li, superoxide anion), lossis radicals (xws li, hydroxyl radicals) thiab siv lawv cov teebmeem los ntawm kev tswj hwm ntawm tes taw qhia cascades [6]. Lawv sai sai thiab tshem tawm yog cuam tshuam los ntawm ntau yam mechanisms. Lawv yog lub teeb yuag thiab diffuse yooj yim hla luv luv [7]. Lub superoxide anion () yog ib qho ua ntej ntawm feem ntau ROSs thiab ib hom kab nruab nrab hauv cov tshuaj tiv thaiv oxidative. Hydroxyl radicals () yog catalyzed los ntawm txo kev hloov pauv hlau, uas yuav nyob rau hauv lem yuav reoxidised los ntawm. Ib qho kev tsis txaus ntseeg ntawm kev tsim ntau dhau ntawm ROS thiab txwv tsis pub muaj kev tiv thaiv antioxidant ua rau muaj ntau yam txheej txheem tseem hu ua "oxidative stress" [8]. Yog tias qhov tsis txaus ntseeg no tuaj yeem kho tau, kev tswj hwm ntawm ntau lub tshuab tiv thaiv yuav raug tswj xyuas. Kev ntxhov siab oxidative cuam tshuam nrog ntau yam kab mob pathological, xws li mob qog noj ntshav, kab mob plawv, kab mob hauv lub paj hlwb, thiab ntshav qab zib [6]. Tsis tas li ntawd, 2, 2-diphenyl-1- picrylhydrazyl (DPPH•) radical yog xim thiab ruaj khov zoo kawg li, thiab yog li ntawd yog ib qho ntawm cov radicals ntau tshaj plaws hauv ntau cov kev tshawb fawb [9]. Kev soj ntsuam DPPH scavenging yog ib qho yooj yim-rau-ua, yog txoj hauv kev los ntsuas tag nrho cov antioxidant peev xwm ntawm botanical lossis tshuaj ntsuab rho tawm [10]. Yog li ntawd, DPPH, superoxide anion, thiab hydroxyl radical scavenging assays tau raug siv los soj ntsuam cov antioxidant muaj peev xwm ntawm txawv extracts ntawmC. tubulosa.
Cov kauj ruam tsim tawm rau cov tshuaj suav tshuaj (TCM) yog qhov nyuaj thiab koom nrog kev txiav, ua, rho tawm, concentration, thiab ziab [11]. Nws yog ib qho tseem ceeb uas cov khoom siv hauv cov khoom siv tshuaj ntsuab (HMs) yuav ploj mus thaum lub sijhawm tsim khoom lag luam zoo li no. PhGs yog tus cwj pwm los ntawm tsawg kawg ib qho glycosyl moiety ntawm lawv cov tub ntxhais, uas txiav txim siab lawv cov khoom. Cov tebchaw no yog dej soluble thiab yooj yim rau rho tawm los ntawm cov txheej txheem ib txwm muaj. Nws tseem yog rooj plaub uas cov tshuab nqus dej tau siv dav hauv kev tsim TCM [12]. Cov dej uas tau muab rho tawm (WEs) yog tsim los rau hauv ntau hom tshuaj xws li ntsiav tshuaj, granules, capsules, thiab mixs. Inevitably, qhov poob ntawm cov khoom xyaw lom tshwm sim thaum lub sij hawm xws li ib tug loj-scale ntau lawm txheej txheem. Qhov no yog vim li cas kev ntsuam xyuas zoo yog ib kauj ruam tseem ceeb thiab yuav tsum tau ua kom ntseeg tau tias cov khoom zoo ib nrab ntawm cov txheej txheem. Qee cov kws tshawb fawb twb tau siv cov khoom seem no los ntawm kev hloov kho cov qauv.Astragalus membranaceousresidue tau huv si los tsim cov polysaccharide uas txhim kho kev paub tsis meej los ntawm kev hloov cov plab microbiota hauv cov nas mob ntshav qab zib [13]. Ib qho nruab nrab polysaccharide muab rho tawm los ntawmCodonopsis pilosularesidue nthuav tawm cov nyhuv hypoglycemic [14]. Ntau cov tshuaj ntsuab residues (HRs) yog tsim nyob rau hauv Suav teb. Kev rov siv dua ntawm HRs tau dhau los ua qhov kev tshawb fawb tshiab thiab tshiab raws li cov txheej txheem TCM tau dhau los ua kom tshiab [15]. Lub peev xwm rau qhov sib txawv ntawm cov khoom antioxidant ntawm ntau yam tshuaj ntsuab thiab lawv cov WEs tau dhau los ua qhov tseem ceeb ntawm peb cov kev xav. Ntxiv rau qhov xav txog WEs ntawmC. tubulosa, nws vam tias yuav txiav txim siab seb HRs puas tuaj yeem ua pov thawj muaj txiaj ntsig zoo li cov neeg sib tw TCMs thiab tshawb nrhiav cov hauv kev rau kev hloov pauv pov tsegC. tubulosacov khoom pov tseg rau hauv cov khoom siv tau.
Kev sib raug zoo ntawm spectrum yog siv los txiav txim siab cov khoom zoo hauv cov khoom sib xyaw ua ke thiab muaj kev cuam tshuam rau sab hauv zoo ntawm cov tshuaj ntsuab. Nws yog indispensable nyob rau hauv tus txheej txheem ntawm modernization thiab internationalization ntawm herbal tshuaj. Txij li thaum kev tshawb fawb txog kev sib raug zoo ntawm cov tshuaj ntsuab yog ua raws li tus ntiv tes chromatographic, yuav tsum muaj txoj hauv kev tsim nyog los tsim cov ntiv tes uas qhia txog cov khoom xyaw tshuaj ntawm cov tshuaj ntsuab. High-performance kua chromatography (HPLC) yog ib qho tseem ceeb analytical txoj kev uas muaj ntau yam zoo, xws li kev sib cais siab, zoo stability, high efficiency, thiab high quantitative precision. PLSR (ib feem tsawg squares regression) ua kom yooj yim cov ntaub ntawv qauv thiab kev txheeb xyuas kev sib raug zoo ntawm ob pawg ntawm qhov sib txawv los ntawm kev siv tus qauv regression [16]. Hauv kev txheeb xyuas kev sib raug zoo ntawm kev sib raug zoo (BCA), cov qhab nia xeem tau cuam tshuam nrog cov tswv yim muaj feem cuam tshuam los tsim cov pov thawj siv tau [17]. RDA (kev txheeb xyuas qhov tsis txaus ntseeg) tau siv los txheeb xyuas thawj cov zej zog microbial ntsig txog kev muaj peev xwm lom neeg tshwj xeeb, tab sis peb siv nws rau kev sib raug zoo spectrum-zoo [18]. Txhawm rau ntsuas qhov sib txheeb coefficients, PLSR thiab BCA tau siv. Cov txiaj ntsig tau raug txheeb xyuas siv RDA los txiav txim siab seb cov qauv twg tsim nyog rau kev kawm txog kev sib raug zoo ntawm kev sib raug zoo nrogC. tubulosa.
Hauv cov txheej txheem ntawm "multicomponent therapeutics, biological network" hauv network pharmacology, peb sim tshawb nrhiav cov hom phiaj ntawm cov kab mob nquag thiab cov kab mob, uas tuaj yeem ua lub luag haujlwm tseem ceeb hauv kev muab cov ntaub ntawv siv rau kev tiv thaiv kab mob.Cistanche tshuaj ntsuabnthuav tawm cov khoom muaj peev xwm ntau thiab ntau lub hom phiaj hauv cov ntawv qhia dhau los [19, 20]. Biomarkers ntawm kev puas tsuaj oxidative cuam tshuam nrog tib neeg cov kab mob tau sau tseg [21]. Ob peb txoj kev tshawb fawb qhia meej txog cov tshuaj tiv thaiv oxidative ntawmC. tubulosa. Yog li ntawd, lub network pharmacology technology tau saws los soj ntsuam bioactive molecules ntawmC. tubulosathiab mechanisms ntawmC. tubulosativ thaiv oxidation.
Hauv txoj kev tshawb no, chromatographic fingerprints thiab antioxidant kev ua ub no ntawm HMs, WEs, thiab HRs los ntawm 11 pawg ntawmC. tubulosatau soj ntsuam ib txhij ntawm HPLC thiab tshuaj tiv thaiv antioxidant. Ib qho kev sib raug zoo ntawm kev sib raug zoo ntawm HPLC cov ntiv tes thiab cov teebmeem antioxidant ntawmC. tubulosatau qhia meej meej los ntawm kev txheeb xyuas kev sib raug zoo. Cov kev tshawb fawb uas twb muaj lawm feem ntau tshaj tawm txog cov khoom tiv thaiv oxidative ntawmC. tubulosatshuaj ntsuab [22–24], tab sis ob peb ntawm lawv hais txog qhov muaj peev xwm antioxidant ntawm HEs. Tsis tas li ntawd, lub luag haujlwm ntawm HRs tej zaum yuav tsis txaus ntseeg, thiab txoj kev tshawb no muab lub sijhawm tshiab los coj kom zoo dua ntawm kev tshawb fawb tshiab hauv daim teb no. Lub hom phiaj ntawm kev tshawb fawb no yog txhawm rau txheeb xyuas cov khoom xyaw tseem ceeb hauv thiab cov haujlwm antioxidant ntawm HMs, WEs, thiab HRs ntawmC. tubulosa. Tom qab ntawd, cov tshuaj chemometrics tau siv los txheeb xyuas qhov kev sib raug zoo ntawm kev sib raug zoo rau HMs, WEs, thiab HRs los ntawm 11 pawg ntawmC. tubulosa. Nov yog thawj zaug uas sib txawv ntawm HMs, WEs, thiab HRs ntawmC. tubulosatau kawm li no. Nyob rau hauv tas li ntawd, lub network pharmacology tsom xam tau ua kom elucidate lub hauv paus mechanisms ntawmC. tubulosahauv kev kho cov kab mob antioxidant.
1. Taw qhia
Cistanche tubulosa(Schenk) R. Wight, ib qho ntawm feem ntau siv tshuaj ntsuab nyob rau hauv lubCistanchetsev neeg, hu ua "Ginseng ntawm suab puam" rau nws cov txiaj ntsig kev noj qab haus huv ntau yam [1]. Kev tshawb nrhiav tshuaj niaj hnub tau tshawb pom tiasCistanchetsev neeg muaj ntau yam tshuaj, xws li antioxidant, anticancer, hypoglycemic, antidepressant, kev txawj ntse txhim kho, thiab cov tshuaj tiv thaiv kab mob [2]. Lub hauv paus chemical pawg hauv Cistancheherb a yog phenylethanoid glycosides (PhGs) [1]. PhGs muab los ntawm Cistanche hom tuaj yeem txhawb nqa cov haujlwm ntawm superoxide dismutase (SOD) thiab glutathione peroxidase (GSH-Px) [3]. Echinacoside, ib tug tswv cuab ntawm PhG tsev neeg, thim rov qab zoo nyob rau hauv vivo oxidative kev nyuaj siab tshwm sim los ntawm high-qabzib noj cov zaub mov ntawm downregulation ntawm cov nitrous oxide system (NOS) kev ua si thiab phospho-eNOS qhia [4]. Lub cev pob txha ntawm PhG muaj phenylethyl cawv thiab glycosyl moieties. PhGs ntseeg tau tias muaj cov haujlwm tseem ceeb vim yog phenolic hydroxyl pawg hauv lawv cov qauv. Kev ua haujlwm antioxidant ntawm PhGs nce nrog cov phenolic hydroxyl pawg [5].
Reactive oxygen hom (ROS) muaj lub luag haujlwm tseem ceeb hauv ntau cov txheej txheem physiological thiab cov txheej txheem tiv thaiv tseem ceeb. Kev nquag raug rau ROS siab ntau tuaj yeem ua rau tsis muaj kev puas tsuaj rau cov protein, lipids, thiab nucleic acids. ROSs tuaj yeem yog cov molecules nruab nrab (xws li, hydrogen peroxide), ions (piv txwv li, superoxide anion), lossis radicals (xws li, hydroxyl radicals) thiab siv lawv cov teebmeem los ntawm kev tswj hwm ntawm tes taw qhia cascades [6]. Lawv sai sai thiab tshem tawm yog cuam tshuam los ntawm ntau yam mechanisms. Lawv yog lub teeb yuag thiab diffuse yooj yim hla luv luv [7] Lub superoxide anion () yog ib tug precursor ntawm feem ntau ROSs thiab ib tug nruab nrab hom nyob rau hauv oxidative tshua. Hydroxyl radicals () yog catalyzed los ntawm txo kev hloov hlau, uas tej zaum yuav reoxidised los ntawm . Ib qho kev tsis sib haum xeeb ntawm kev tsim ntau dhau ntawm ROS thiab txwv kev tiv thaiv antioxidant ua rau ntau yam txheej txheem tseem hu ua "oxidative stress" [8]. Yog tias qhov tsis txaus ntseeg no tuaj yeem kho tau, kev tswj hwm ntawm ntau lub tshuab tiv thaiv yuav raug tswj xyuas. Oxidative stress yog koom nrog ntau yam kab mob, xws li mob qog noj ntshav, kab mob plawv, kab mob hauv lub paj hlwb, thiab ntshav qab zib [6]. Tsis tas li ntawd, 2, 2-diphenyl-1- picrylhydrazyl (DPPH•) radical yog xim thiab ruaj khov zoo kawg li, thiab yog li yog ib qho ntawm cov radicals ntau tshaj plaws hauv ntau cov kev tshawb fawb [9]. Kev soj ntsuam DPPH scavenging yog ib qho yooj yim-rau-ua, yog txoj hauv kev los ntsuas tag nrho cov antioxidant peev xwm ntawm botanical lossis herbal extracts [10]. Yog li ntawd, DPPH, superoxide anion, thiab hydroxyl radical scavenging assays tau raug siv los soj ntsuam cov antioxidant muaj peev xwm ntawm txawv extracts ntawmC. tubulosa.
Cov kauj ruam tsim tawm rau cov tshuaj suav tshuaj (TCM) yog qhov nyuaj thiab koom nrog kev txiav, ua, rho tawm, concentration, thiab ziab [11]. Nws yog ib qho tseem ceeb uas cov khoom siv hauv cov khoom siv tshuaj ntsuab (HMs) yuav ploj mus thaum lub sijhawm tsim khoom lag luam zoo li no. PhGs yog tus cwj pwm los ntawm tsawg kawg ib qho glycosyl moiety ntawm lawv cov tub ntxhais, uas txiav txim siab lawv cov khoom. Cov tebchaw no yog dej soluble thiab yooj yim rau rho tawm los ntawm cov txheej txheem ib txwm muaj. Nws kuj tseem yog cov ntaub ntawv uas cov tshuab nqus dej tau siv dav hauv kev tsim TCM [12]. Cov dej uas tau muab rho tawm (WEs) yog tsim los rau hauv ntau hom tshuaj xws li ntsiav tshuaj, granules, capsules, thiab mixs. Inevitably, qhov poob ntawm cov khoom xyaw lom tshwm sim thaum lub sij hawm xws li ib tug loj-scale ntau lawm txheej txheem. Qhov no yog vim li cas kev soj ntsuam zoo yog ib kauj ruam tseem ceeb thiab yuav tsum tau ua kom ntseeg tau tias cov khoom lag luam semifinished zoo los ntawm cov txheej txheem. Qee cov kws tshawb fawb twb tau siv cov khoom seem no los ntawm kev hloov kho cov qauv. Astragalus membranaceus residue tau ua kom huv los tsim cov polysaccharide uas txhim kho kev paub tsis meej los ntawm kev hloov plab microbiota hauv cov nas mob ntshav qab zib [13]. Ib qho nruab nrab polysaccharide muab rho tawm los ntawm Codonopsis pilosula residue tau nthuav tawm cov nyhuv hypoglycemic [14]. Ntau cov tshuaj ntsuab residues (HRs) yog tsim nyob rau hauv Suav teb. Kev rov siv dua ntawm HRs tau dhau los ua qhov kev tshawb fawb tshiab thiab tshiab raws li cov txheej txheem TCM tau dhau los ua tshiab [15]. Lub peev xwm rau qhov sib txawv ntawm cov khoom antioxidant ntawm ntau yam tshuaj ntsuab thiab lawv cov WEs tau dhau los ua qhov tseem ceeb ntawm peb cov kev xav. Ntxiv nrog rau kev txiav txim siab WEs ntawm C. tubulosa, nws vam tias yuav txiav txim siab seb HRs tuaj yeem ua pov thawj muaj txiaj ntsig zoo li cov neeg sib tw TCMs thiab tshawb nrhiav cov hauv kev los hloov cov khoom pov tseg C. tubulosa rau hauv cov khoom siv tau.
Kev sib raug zoo ntawm spectrum yog siv los txiav txim siab cov khoom zoo hauv cov khoom sib xyaw ua ke thiab muaj kev cuam tshuam rau sab hauv zoo ntawm cov tshuaj ntsuab. Nws yog indispensable nyob rau hauv tus txheej txheem ntawm modernization thiab internationalization ntawm herbal tshuaj. Txij li thaum lub spectrum-zoo kev sib raug zoo kev tshawb fawb ntawm cov tshuaj ntsuab yog raws li nyob rau hauv lub c
Daim duab 1 Cov txheej txheem ntawm kev ua cov khoom siv raw ntawm C. tubulosa rau hauv ntau hom
2.3. HPLC Kev Kho Mob ntawm Cistanche
Chromatography tau ua tiav siv Vanquish™ Horizon UHPLC (Thermo Fisher Scientific, Massachusetts, USA) nrog lub ntsuas hluav taws xob ultraviolet thiab ACQUITY UPLC BEH C18 kem (2.1 hli × 100 hli, 1.7 μm) ntawm 3 0 degree. Cov theem txawb muaj 0.1 feem pua phosphoric acid tov (A) hauv acetonitrile (B) hauv gradient elution hom raws li nram no: 0–8 min 5 feem pua – 13 feem pua B, 8-9 min 13 feem pua – 15 feem pua B, 9 -19 min 15 feem pua B, 19–20 min 15 feem pua -5 feem pua B, thiab 20–26 min 5 feem pua B. Tus nqi ntws yog 0.3 mL / min, cov qauv txhaj tshuaj yog 1 µL, thiab lub ntsuas wavelength yog teem rau 238 nm.
2.4. Kev Npaj Cov Qauv thiab Cov Qauv rau HPLC Kev Ntsuam Xyuas Cistanche
2.4.1. Kev npaj cov qauv kev daws teeb meem
Cov qauv kev daws teeb meem tau npaj raws li Zhen li al. nrog kev hloov kho [28]. Cistanche tubulosa tshuaj ntsuab thiab cov dregs tau muab tso rau hauv hmoov (65 mesh). Cov qauv pulverized (1. (250 W, 40 kHz). Cov kev daws teeb meem tau lim los ntawm 0.22-μL microporous membrane. Tom qab ntawd, cov qauv ntawm WEs (0.2 g) tau muab rho tawm nrog 50 mL ntawm 50 feem pua aqueous methanol hauv ib da dej ultrasonic rau 30 min (250 W, 40 kHz). Cov kev daws teeb meem tau lim los ntawm 0.22-μL microporous membrane.
2.4.2. Kev npaj cov txheej txheem daws teeb meem ntawm Cistanche
Cov nqi tsim nyog ntawm tsib qhov sib xyaw ua ke tau yaj hauv 50 feem pua aqueous methanol thiab tom qab ntawd lim los ntawm 0.22-μL microporous membrane kom tawm los ntawm cov txheej txheem sib xyaw. Thaum ntxiv 50 feem pua aqueous methanol rau ib qho 10-mL ntim ntim, cov txheej txheem sib xyaw muaj cov geniposidic acid,echinacoside cov tshuaj, lus verbascoside, tubulosideA, thiab isoacteoside. Pure reference compound solutions raug txhaj rau hauv HPLC system rau kev soj ntsuam zoo, thiab lawv lub sijhawm khaws cia tau raug kaw. Sib piv cov sij hawm khaws cia tso cai rau kev txheeb xyuas cov ntsiab lus.
2.5. Methodology Validation ntawm Cistanche
2.5.1. Precision, Reproducibility, thiab Stability
Cov hmoov ntawm HM1 tau npaj raws li tau piav qhia hauv Tshooj 2.4. Cov txheej txheem precision raug soj ntsuam siv rau kev txhaj tshuaj ua tiav ntawm ib qho kev daws teeb meem, thaum lub sijhawm rov tsim dua tau kwv yees los ntawm kev ua rau 6 qhov rov ua dua ntawm cov qauv. Kev ntsuam xyuas kev ruaj ntseg tau ua los ntawm kev rov ua cov tshuaj txhaj tshuaj ntawm ib qho qauv tshuaj uas tau khaws cia ntawm 15 degree rau 0, 2, 4, 8, 12, thiab 24 h.
2.5.2. Linearity
Cov kev daws teeb meem sib xyaw nrog qhov sib txawv gradient concentrations tau txhaj thiab tshuaj xyuas. Cov concentration ntau rau geniposidic acid, echinacoside, verbascoside, tubuloside A, thiab isoacteoside yog {{0}}.0011–{10}}. 3414 mg/mL, 0. raws. Kev ntsuas nkhaus rau txhua tus qauv tau txais los ntawm kev txiav txim siab qhov sib txheeb ntawm thaj chaw ncov (y) thiab concentration (x, mg / mL) siv tus qauv qis tshaj plaws squares.
2.5.3. Qauv rov qab
Cov qauv rov qab tau raug tshawb xyuas los ntawm kev ntxiv qhov tseeb ntawm cov qauv kev daws teeb meem rau 0.5 g ntawm HM1 qauv hmoov. Cuaj qhov qauv tau npaj ua ke raws li Tshooj 2.4. Qhov nruab nrab tus qauv rov qab los ntawm txhua yam tau txiav txim siab.
2.5.4 ib. Kev txiav txim siab
Raws li tau piav qhia hauv Tshooj 2.4, HMs, Wes, thiab HRs tau npaj ua ke. Cov qauv kev daws teeb meem no tau txhaj tshuaj raws li cov xwm txheej chromatographic tau piav qhia hauv Tshooj 2.3. Lub ncov cheeb tsam raug kaw thiab cov ntsiab lus raug xam. Cov ntaub ntawv tau txheeb xyuas siv GraphPad Prism 8 (GraphPad Software, California, USA). Cov txiaj ntsig tau suav nrog siv ib txoj kev tsom xam ntawm qhov sib txawv ua raws li txoj kev Tukey. tau suav hais tias tseem ceeb heev.
2.6. Fingerprint Tsim thiab Kev Ntsuas ntawm Cistanche
HPLC chromatographic cov ntaub ntawv tau tso tawm los ntawm Chromeleon 7.2.8 Software (Thermo Fisher Scientific, Massachusetts, USA) hauv CDF thiab TXT hom. HM-WE, WE-HR, thiab HM-HR qhov zoo sib xws tau suav nrog siv qhov kev ntsuam xyuas zoo sib xws uas tsim los rau cov ntiv tes chromatographic hauv TCM Software (Version 2004A). HPLC ntiv tes tau kos siv Keeb Kwm 2021 Software (OriginLab, Massachusetts, USA).
2.7. Antioxidant Kev Ntsuas Kev Ntsuas ntawm Cistanche
Cov txheej txheem ntsuas rau qib antioxidant tau ua raws li cov lus qhia hauv ntau yam khoom siv. Qhov ntsuas qhov nqus tau raug ntsuas siv Shimadzu UV-2600i (Shimadzu, Nyiv). Txhua tus qauv tau khiav hauv triplicate, thiab cov ntaub ntawv nruab nrab raug kaw. Ntau cov qauv ntsuas thiab lawv qhov sib thooj absorbance (A) qhov tseem ceeb tau sau tseg. 50 feem pua inhibiting concentration (IC50) tau suav nrog GraphPad Prism 8 (GraphPad Software, California, USA).
Lub cev qhuav dej ethanol tau siv los kho rau xoom.
2.7.2. Assay
Ua ntej, 0.1 g ntawm HM, WE, thiab HR hmoov tau muab rho tawm siv 1 mL ntawm 80 feem pua aqueous ethanol tov. Cov khoom tshwm sim tau sonicated rau 30 min thiab centrifuged ntawm 5 min. Ib qho piv txwv 100 μL tau ntxiv rau hauv cov kua dej ua haujlwm (Table S2). Tom qab cov tshuaj tiv thaiv tau tso cai mus rau ntawm 37 degree rau 10 min, qhov absorbance ntawm 570 nm tau kaw thiab hloov mus rau radical scavenging kev ua () raws li nram no:
Cov dej huv huv tau siv los kho kom xoom.
2.8. Kev txheeb xyuas cov ntaub ntawvntawm Cistanche
2.8.1 ib. PLSR Analysis
Lub ntsiab chromatographic ncov cheeb tsam tau txais kev pab raws li qhov sib txawv ntawm kev ywj pheej (X) thiab qib ntawm cov tshuaj tiv thaiv antioxidant rau ntau qhov kev ntsuam xyuas yog qhov sib txawv (Y). PLSR qauv tau ua tiav siv Unscrambler X 10.4 Software (CAMO Software, Bangalore, India). Qhov hnyav regression coefficients qhia txog kev sib raug zoo ntawm thaj chaw ncov thiab qib kev ua haujlwm antioxidant, thiab cov nyom regression coefficients txhais cov qauv sib npaug.
2.8.2 ib. BCA Analysisntawm Cistanche
Peak thaj chaw yog qhov sib txawv ntawm kev ywj pheej (X), thiab cov qib antioxidant rau ntau qhov kev ntsuam xyuas tau raug kho raws li qhov sib txawv (Y). Tom qab ntawd, BCA ntawm X thiab Y tau txheeb xyuas siv tus qauv Pearson. Cov txheej txheem no tau ua tiav siv SPSS cov ntaub ntawv txheeb xyuas software (SPSS rau Windows 26.0, SPSS Inc., San Francisco, USA).
2.8.3 ib. Heatmap Chart, Venn Chart, thiab RDAntawm Cistanche
Heatmap kab kos tau kos siv GraphPad Prism 8 (GraphPad Software, California, USA). Raws li cov qauv PLSR thiab BCA, qhov kev sib raug zoo ntawm thaj chaw ncov thiab lub peev xwm antioxidant ntawm 11 peaks tau muab teev los ntawm loj mus rau me me, thiab cov qib tsib tau sau tseg. Cov peaks no tau siv los kos duab Venn hauv online (https://bioinformatics.psb.ugent.be/webtools/Venn/). Cov ntaub ntawv rau ntau qhov chaw ncov thiab qib antioxidant rau peb hom C. tubulosa tau pom los ntawm RDA. Cov kev xeem RDA tau ua tiav siv CANOCO Software (Biometris-Plant Research International, Wageningen, Netherlands).
2.9. Network Pharmacology Analysis ntawm Cistanche
2.9.1 ib. Kev tshuaj ntsuam xyuas cov khoom xyaw nquag ntawm C. tubulosa
Tag nrho cov tshuaj lom neeg ntawm C. tubulosa tau txais kev siv tshuaj suav tshuaj suav tshuaj (TCMSP, https://www.tcmsp-e.com/). Qhov kev tshuaj ntsuam xyuas ntawm txhua yam tshuaj lom neeg tau teem tseg raws li qhov ncauj bioavailability (OB) Ntau dua lossis sib npaug rau 30 feem pua thiab cov tshuaj zoo li (DL) Ntau dua lossis sib npaug rau 0.18, raws li. InChIKey ntawm cov khoom xyaw bioactive tau sau los ntawm PubChem database (https://pubchem.ncbi.nlm.nih.gov/). Cov hom phiaj protein ntawm cov tshuaj nquag tau raug tshuaj xyuas los ntawm SwissTargetPrediction database (https://www.swisstargetprediction.ch/). Cov npe lub hom phiaj tau hloov mus rau hauv cov npe noob siv UniPort protein database (https://www.uniprot.org/).
2.9.2 ib. Kev tsim kho ntawm Cov Txheej Txheem-Target Network
Lo lus tseem ceeb "antioxidant" tau siv los tshawb nrhiav cov hom phiaj ntsig txog kab mob ntawm GeneCards database (https://www.genecards.org/) thiab OMIM database (https://omim.org/). Cov kev sib tshuam ntawm cov noob ntawm cov khoom nquag thiab cov hom phiaj ntsig txog kab mob tau pom los ntawm kev siv daim duab Venn online (https://bioinformatics.psb.ugent.be/webtools/Venn/). Lub hom phiaj bioactive ingredient ntawm C. tubulosa tau mapped rau lub hom phiaj genes siv Cytoscape 3.9.1 software (https://cytoscape.org/) rau kev tsim cov tivthaiv-target (CT) network.
2.9.3 ib. Gene Ontology thiab Kyoto Encyclopedia ntawm Genes thiab Genomes Enrichment Analyzes
Gene ontology (GO) enrichment in biological processes (BP), cellular component (CC), and molecular function (MF), thiab kyoto encyclopedia of genes and genomes (KEGG) pathway enrichment tau txheeb xyuas online siv Metascape database (https:// www.metascape.org/) nrog rau qhov "Homo sapiens" chaw. Daim duab npuas pom pom thiab GO histogram tau tsim hauv online (https://www.bioinformatics.com.cn/).
2.9.4 ib. Tsim cov Protein-Protein Interaction thiab Component-Target-Pathway Networks
The overlapping antioxidation-related and predicted targets from active components were used to construct a protein-protein interaction (PPI) using the STRING database (https://stringdb.org/). The conditions were set as described by Xin et al. [29]. The PPI network was visualized using the Cytoscape software. Degree centrality (DC), betweenness centrality (BC), and closeness centrality (CC) were calculated through the "network analysis" function. DC, BC, and CC were set as >100, >0.03, and >0.3, raws. Raws li KEGG txoj hauv kev thiab lub hom phiaj cov noob, Cytoscape software tau siv los tsim kom muaj lub hom phiaj-txoj kev (CTP) network.
