Antioxidant Activity, Phenolic Profile, Thiab Nephroprotective Potential Ntawm Anastatica Hierochuntica Ethanolic Thiab Aqueous Extracts Tawm tsam CCl4-Induced Nephrotoxicity hauv nas

Hu rau: ali.ma@wecistanche.com

Tariq I. Almundarij et al

Abstract:Kaff-e-Maryam (Anastatica hierochuntica L.) feem ntau siv los kho ntau yam teeb meem kev noj qab haus huv, tshwj xeeb tshaj yog kom yooj yim rau kev yug menyuam thiab txo cov kab mob ntsig txog kev sib deev. Txoj kev tshawb no tsom los ntsuas qhov cuam tshuam ntawm A. hierophantic ethanolic (KEV), thiab aqueous (KAE) extraction CCl 4-induced oxidative stress thiab nephrotoxicity hauv nas siv cov cim biochemical raulub raumkev ua haujlwmthiab antioxidant raws li txoj cai raws li kev kuaj histopathological ntawm lub raum cov ntaub so ntswg.A. hierophantic muaj 67.49 mg GAE g-1 ntawm tag nrho cov phenolic compounds (TPC), 3.51 µg g-1 ntawm tag nrho carotenoids (TC), thiab 49.78 thiab 17.45 mg QE g-1 ntawm tag nrho cov flavonoids (TF) thiab tag nrho flavonols (TFL), raws. Nws ua rau 128.71 µmol ntawm TE g-1 ntawm DPPH-RSA thiab 141.92 µmol ntawm TE g-1 ntawm ABTS-RSA. A. hierophantic nthuav qhia superiorkev ua haujlwm antioxidantlos ntawm inhibiting linoleic acid radicals thiab chelating oxidation hlau. Kev txheeb xyuas HPLC tau ua rau 9 thiab 21 phenolic acids thiab 6 thiab 2 flavonoids hauvKEVthiab KAE nrog qhov tseem ceeb ntawm sinapic thiab syringic acids, feem. Intramuscular txhaj ntawm vit. E plus Se thiab kev tswj hwm qhov ncauj ntawm KEE, KAE, thiab KEE ntxiv rau KAE ntawm 250 mg kg-1 lub cev hnyav tau rov qab ua cov ntshav creatinine, urea, K, tag nrho cov protein, thiab qib albumin. Tsis tas li ntawd, lawv txo qis malondialdehyde (MOD), rov qab txo qis glutathione (GSH), thiab txhim kho superoxide dismutase (SOD) qib. KEE, KAE, thiab KEE plus KAE tiv thaivlub raumlos ntawm CCl4-nephrotoxicity raws li lawv feem ntau attenuated induced oxidative stress. Tag nrhonephron uaseem yog kwv yees li 83.27 feem pua, 97.62 feem pua, thiab 78.85 feem pua ​​rau KEE, KAE, thiab KEE ntxiv rau KAE, feem. Ob leeg vit. E plus Se thiab A. hierophantic extracts attenuated the histopathological alteration in CCl4-kho nas. Hauv kev xaus, A. hierophantic, tshwj xeeb tshaj yog KAE, muaj peev xwm rov qab oxidative stability thiab txhim khoraumua haujlwm tom qab CCl4mob raum raug mobzoo dua KEE. Yog li, A. hierophantic muaj peev xwm los ua tus neeg sawv cev kho mob muaj txiaj ntsig hauv kev kho mob ntawm cov tshuaj tiv thaiv nephrotoxicity.

Ntsiab lus:Kaf-e-Maryam; polyphenols; bioactivity; theem nrab metabolites;lub raum cim; antioxidant enzymes;lub raumkev ua haujlwm tsis zoo

Cistanche NZ tiv thaiv kab mob raum, nyem qhov no kom tau txais cov qauv

1. Taw qhia

Kab mob raumyog qhov thib 9 ua rau tuag nrog ntau dua 1 ntawm 7, uas yog, 15 feem pua ​​​​ntawm Asmeskas cov neeg laus lossis 37 lab tus tib neeg, kwv yees tias muajmob raum mob(CKD) [1]. Remarkably, feem ntau ua rau CKD raws li sau tseg nyob rau hauv 2015 yog ntshav qab zib mellitus ua raws li ntshav siab thiab glomerulonephritis [2]. Lwm yam ua rau CKD muaj xws li idiopathic (feem ntau txuam nrog raum me me ntawm lub raum ultrasound) [3]. Yav dhau los, CCl4 tau siv rau hlau degreasing, qhuav tu, ntaub spotting, tua hluav taws, thiab grain fumigation [4]. Nws ua rau mob hnyav hauv daim siab, ntsws, thiab cov qog ntshav thiab hauv cov ntshav los ntawm kev tsim cov dawb radicals [5]. Raws li kev tshawb pom ntawm Ogeturk li al. [6], raug rau cov kuab tshuaj no ua rau mob hnyav thiab mob raum puas. Dawb radical-induced lipid peroxidation ntseeg tau tias yog ib qho ntawm cov tseem ceeb ua rau cell membrane puas, ua rau ntau yam pathological mob [7]. Cov tiam ntawm reactive radicals tau cuam tshuam rau hauv CCl 4-induced nephrotoxicity, uas yog koom nrog hauv lipid peroxidation thiab tsub zuj zuj ntawm cov proteins uas tsis ua haujlwm, ua rau raug mob raum [8]. Amazing, ib txwm siv cov nroj tsuag tshuaj tau loj hlob nyob rau hauv xyoo tas los no, thiab ntau qhov kev tshawb fawb tau lees paub lawv txoj haujlwm kho mob tiv thaiv ntau yam mob [9–12].

2. Cov ntaub ntawv thiab cov txheej txheem

2.1. Kev npaj ua qauv

cov qauv ntawm Kaff-e-Maryam (A. hierophantic L.) cog tau yuav los ntawm ib lub khw muag khoom hauv nroog Buraydah, cheeb tsam Qassim, Saudi Arabia. Cov khoom siv cog tau raug lees paub los ntawm Lub Tsev Haujlwm Saib Xyuas Kev Tsim Khoom thiab Kev Tiv Thaiv, Tsev Kawm Qib Siab Kev Ua Liaj Ua Teb thiab Veterinary Medicine, Qassim University, Saudi Arabia. Cov qauv raug ntxuav nrog dej huv kom tshem tawm cov xuab zeb thiab av los ntawm nplooj thiab tom qab ntawd cov khoom siv cua qhuav (ntawm 28 ± 1 ◦C rau 48 h.) yog cov hmoov txhuam thiab khaws cia rau hauv cov hnab polyethylene opaque ntawm 4 ± 1 ◦C mus txog thaum siv.

2.2. Kev npaj ntawm Ethanolic thiab Aqueous Extracts

Kwv yees li 200 g ntawm qhuav A. hierophantic tau muab rho tawm nrog 300 mL 70 feem pua ​​​​ethanol hauv Soxhlet extractor los npaj ethanolic extraction (KEE). Cov extract yog concentrated los ntawm ib tug teb evaporator ntawm 40 ◦C kom evaporate cov kuab tshuaj seem, ces mus rau qhuav nyob rau hauv ib tug N2 kwj. Lub aqueous extraction (KAE) tau ua raws li tau piav qhia los ntawm Asuzu [23] nrog kev hloov kho me me. Ob puas grams ntawm qhuav nroj tsuag cov khoom tau ntxiv rau 500 mL ntawm kub sterile distilled dej. Tom qab ntawd cov sib tov yog shaken zoo thiab tso cai rau sawv ntsug rau 1 h. Tom qab ntawd ib lub reflux condenser txuas mus rau lub hwj thiab tom qab ntawd rhaub kom txog thaum boiling maj mam rau 10 min, txias, shaken zoo, thiab lim los ntawm Whatman No. 1 lim ntawv. Cov filtrate tau evaporated los ntawm lub teb evaporator, ces mus rau dryness nyob rau hauv ib tug N2stream. Cov dej cawv thiab aqueous extracts (250 mg mL-1) tau tsim tshiab hauv cov dej distilled los siv rau kev tswj qhov ncauj.

2.3. Tag nrho cov ntsiab lus Phenolic (TPC)

TPC cov ntsiab lus ntawm A. hierophantic tau txiav txim siab raws li txoj kev hloov kho los ntawm Bettaieb li al. [24]. Cov txiaj ntsig tau muab piv rau ib qho kev qhia ntawm cov kab mob gallic acid (GA) tus qauv nkhaus ua nyob rau hauv thaj tsam ntawm 50–500 mg mL−1 (R2=0.99), thiab TPC raug xam raws li mg ntawm gallic acid sib npaug (GAE ) ib gram ntawm A. hierophantic (mg ntawm GAE g-1 ).

2.4. Tag nrho Carotenoids (TC), Total Flavonoids (TF), thiab Tag Nrho Flavonols (TFL)

Raws li qhia los ntawm Al-Qabba et al. [10], 5 g ntawm A. hierophantic tau muab rho tawm ntau zaus nrog acetone thiab roj av ether sib tov (1: 1, v / v). Tag nrho cov ntsiab lus carotenoids (TC) tau txiav txim siab spectrophotometrically ntawm 451 nm. TC tau qhia tias mg g-1 DW. Cov ntsiab lus TF ntawm A. hierophantic tau soj ntsuam raws li cov txheej txheem piav qhia los ntawm Mohdaly li al. [25]. Cov ntsiab lus TF raug xam raws li mg quercetin sib npaug (QE) ib 100 g-1 DW. Hauv tib lub ntsiab lus, TFL cov ntsiab lus tau ua tiav [26]. Lub absorbance ntawm 440 nm tau sau tseg, thiab TFL raug xam raws li mg quercetin sib npaug (QE) ib 100 g-1 DW.

2.5. Antioxidant Capacity Determination

DPPH radical scavenging assay: RSA raug kuaj spectrophotometrically nyob ntawm qhov bleaching ntawm DPPH ntshav-xim kua raws li cov txheej txheem hloov los ntawm Lu li al. [27]. Lub peev xwm antiradical tau nthuav tawm raws li µmol ntawm Trolox equivalents (TE) ib gram ntawm A. hierophantic (µmoL TE g−1 ). ABTS radical scavenging kev ua ub no: RSA ntawm A. hierophantic tiv thaiv ABTS radicals tau sim los ntawm txoj kev yoog ntawm Lu li al. [27]. Cov txiaj ntsig tau qhia tias µmoL TE g-1. -carotene-linoleic acid bleaching assay: Qhov feem pua ​​​​ntawm cov tshuaj tua kab mob antioxidant ntawm A. hierophantic tau soj ntsuam nyob rau hauv cov ntsiab lus ntawm -carotene bleaching piv rau butylated hydroxyanisole (BHA) thov kev hloov kho spectrophotometric raws tu qauv tsim los ntawm Koleva li al. [28]; Cov txiaj ntsig tau muab raws li BHA feem pua. Chelating kev txiav txim ntawm A. hierophantic ntawm ferrous ions: Kev ua haujlwm chelating ntawm A. hierophantic tau ntsuas raws li kev cai los ntawm Zhao li al. [29]. Qhov inhibition feem pua ​​​​ntawm ferrozine-Fe2 ntxiv rau kev tsim cov hlau chelates tau ntsuas thiab nthuav tawm li mg mL-1 thaum ethylenediaminetetraacetic acid (EDTA) raws li kev tswj xyuas zoo.

2.6. Polyphenolic Compound Fractionation ntawm A. hierophantic Aqueous thiab Ethanolic Extracts

Kev txiav txim siab ntawm polyphenols los ntawm ethanolic thiab aqueous rho tawm tau ua los ntawm HP1100 (Agilent Technologies, Palo Alto, CA, USA) HPLC system nruab nrog ib qho pib-sampler, quaternary twj, thiab diode array detector (Hewlett Packard 1050) siv ib kem (Altima C18 150 × 4.6 mm, 5 µm) nrog 5 mm Altima C18 tus neeg saib xyuas kem (Alltech, Nettetal, Germany) raws li Goupy li al. [30]. Cov kuab tshuaj siv tau yog gradient ntawm A (acetic acid 2.5%), B (acetic acid 8%), thiab C (acetonitrile). Cov kuab tshuaj sib xyaw yog 1 mL min-1, thiab kev sib cais tau ua tiav ntawm 35 ◦C. Lub ntim ntim yog 10 µL. Phenolic tebchaw raug soj ntsuam los ntawm lwm tus qauv calibration thiab qhia raws li mg g-1 DW ntawm equivalents (ntxiv rau )-catechin rau flavan-3-ols thiab sib npaug coumarin rau polar aromatic compounds. Qhov sib txawv ntawm 8 feem pua ​​​​yog txiav txim siab ntawm tsib qhov kev rho tawm ntawm phenolics los ntawm tib tus qauv. Tag nrho cov txiaj ntsig yog qhov txhais tau tias ntawm kev txhaj tshuaj. Polyphenols thiab lawv cov derivatives tau txheeb xyuas thiab ntsuas ntawm 280 thiab 320 nm, thaum flavonoids tau txheeb xyuas ntawm 360 nm.

2.7. Kev sim tsim

Txhua qhov kev sim tau pom zoo los ntawm Pawg Neeg Saib Xyuas Kev Ncaj Ncees Tsiaj (IAEC) ntawm QU (No. {{0}}), KSA, uas yog tswj hwm los ntawm Lub Hom Phiaj ntawm Kev Tswj thiab Kev Saib Xyuas Kev Ua Haujlwm ntawm Tsiaj (CPCSEA) Pawg neeg nyob rau hauv National Committee of Bioethics (NCBE), Ua raws li cov kev cai ntawm txoj cai ntawm kev coj ncaj ncees ntawm kev tshawb fawb ntawm cov tsiaj nyob. Tag nrho ntawm 36 tus txiv neej albino nas tau siv rau hauv txoj kev tshawb fawb tam sim no thiab muab faib ua 6 pawg ntawm 6 tus tsiaj txhua tus thiab kho raws li hauv qab no: Pawg I (Control) tau txais kev txhaj tshuaj intraperitoneal (ip) ntawm cov roj txiv roj (1.0 mL kg−1 ob zaug ib lub lim tiam) thiab 0.5 mL dej distilled ntawm qhov ncauj / ib hnub rau 21 hnub ua ke. Pawg II tau txais kev txhaj tshuaj ip ntawm qhov sib tov tshiab ntawm qhov sib npaug ntawm CCl4 thiab cov roj txiv roj (ntawm ib koob ntawm 1.0 mL kg−1 ob zaug ib lub lim tiam) thiab 0.5 mL distilled dej ntawm qhov ncauj / txhua hnub raws li Al-Qabba et al. [10] nrog kev hloov kho me me. Pawg III (paub pawg) tau txais kev txhaj tshuaj intramuscular (im) ntawm 50 mg kg-1 vit. E plus Se (Selepherol, Vetoquinol Co., Magny-Vernois, Fabkis) ob zaug ib lub lim tiam, raws li Asuka li al. [31] thiab El-Desoky et al. [32], thiab 0.5 mL dej distilled ntawm qhov ncauj / txhua hnub. Pab pawg IV tau ua qhov kev sim thiab tau txais 250 mg kg-1 ntawm KEE qhov ncauj / txhua hnub nrog rau CCl4 ip ob zaug hauv ib lub lis piam. Pawg V tau txais 250 mg kg-1 ntawm KAE qhov ncauj / txhua hnub nrog rau CCl4 ip ob zaug hauv ib lub lis piam. Pawg VI tau txais 250 mg kg-1 ntawm KEE ntxiv rau KAE (1: 1) qhov ncauj / txhua hnub nrog rau CCl4 ip ob zaug hauv ib lub lis piam. Nees nkaum plaub teev tom qab kev kho mob zaum kawg (hnub 21), cov nas tsuag tau tshuaj loog los ntawm kev sib xyaw (cawv: chloroform: ether, 1: 2: 3). Cov ntshav kuaj ntshav los ntawm lub plawv dhia tau sau rau txhua tus tsiaj, thiab cov ntshav tau muab cais los ntawm centrifugation ntawm 4000 rpm rau 10 min thiab khaws cia ntawm −20 ◦C rau kev kuaj biochemical.

2.7.1 ib. Lub raum Biochemical Analysis

Serum creatinine, urea, tag nrho cov protein, thiab albumin concentrations tau txiav txim los ntawm automated spectrophotometric txoj kev (BM/Hitachi autoanalyzer-911; Boehringer Mannheim, Lub teb chaws Yelemees) raws li cov lus qhia ntawm lub khw. Potassium qib tau txiav txim siab los ntawm nplaim taws photometry ntawm 766 nm.

2.7.2 ib. Kev kwv yees ntawm lub raum Antioxidant Activity

Tom qab sau cov ntshav kuaj, cov tsiaj ntawm txhua pab pawg tau muab txi; txoj cailub raumtau cais tawm sai sai thiab yaug nrog dej khov-txias saline. Tom qab ntawd cov ntaub so ntswg yog txiav, rinsed nyob rau hauv txias saline, blotted qhuav, thiab muab tso rau hauv dej khov tam sim ntawd. Siv cov ntaub so ntswg hluav taws xob homogenizer, feem ntawm cov ntaub so ntswg (1.0 g) tau hnyav thiab homogenized nrog 9 ntim ntawm cov dej khov txias 0.05 M phosphate tsis nyob ntawm pH 7.4. Cov khib nyiab ntawm tes raug tshem tawm los ntawm centrifugation ntawm 12, 000 rpm (4 ◦C) rau 20 min los sau cov supernatants rau kev txiav txim siab ntawm malondialdehyde (MDA) concentration [33], superoxide dismutase (SOD) kev ua haujlwm [34], thiab txo glutathione (GSH) cov ntsiab lus [35]. Protein concentration nyob rau hauvraumhomogenate tau txiav txim siab siv Bradford txoj kev [36].

2.7.3 ib. Nephroprotection feem pua

nephroprotection (F) feem pua ​​​​ntawm vit. E plus Se, KEE, KAE, thiab KEE plus KAE raug xam rau txhua qhov biochemical parameter cais raws li Wakchaure li al. [37] siv cov kab zauv hauv qab no:

qhov twg T=txhais tau tias tus nqi ntawm pab pawg kho mob, C=txhais tau tias tus nqi ntawm pawg tswj qhov zoo, thiab N= txhais tau tias tus nqi ntawm pawg tswj tsis zoo. Ntxiv mus, tag nrho nephroprotection feem pua ​​(TFP feem pua) tau muab piv rau vit. E plus Seas hauv qab no:

2.7.4 ib. Kev tshawb fawb Histopathological

Cov qauv kev txiav txim siab tau sau los ntawm sab laugraumntawm cov nas sib cais thiab kho hauv 10 feem pua ​​​​ntawm formalin saline rau 24 teev. Ntxuav nrog kais dej yog ua raws li lub cev qhuav dej nrog serial dilutions ntawm cawv (methyl, ethyl, thiab absolute ethyl). Cov qauv raug tshem tawm hauv xylene thiab muab tso rau hauv paraffin rau 24 teev ntawm 56 ◦C hauv qhov cub cua kub. Paraffin muv wax cov ntaub so ntswg tau npaj rau ntu ntawm 4-micron thickness siv lub sled microtome. Cov ntaub so ntswg tau sau rau ntawm cov iav slides, deparaffinized, thiab stained nrog hematoxylin thiab eosin rau kev soj ntsuam tsis tu ncua nyob rau hauv ib lub teeb hluav taws xob microscope [38].

2.8. Kev txheeb cais

Cov txiaj ntsig tau pom raws li qhov ntsuas ± tus qauv yuam kev (SE). Qhov tseem ceeb ntawm qhov sib txawv ntawm cov txhais tau tias hauv ntau pab pawg tau tshuaj xyuas siv ib txoj kev tsom xam ntawm qhov sib txawv (ANOVA) ua raws li Duncan qhov kev sim, thiab p-tus nqi ntawm cov txhais tau tias tau muab rau ntawm p < 0.05="" qib="" [39="">

3. Cov txiaj ntsig

3.1. Phytochemicals thiab Antioxidant Capacity ntawm A. hierophantic

Kev txheeb xyuas ntau ntawm A. hierophantic phytochemicals thiab lwm yam tshuaj tiv thaiv antioxidant siv DPPH thiab ABTS radical scavenging, -carotene-linoleic acid bleaching kev ua ub no, thiab chelating peev xwm (CA) tau ua. Raws li pom tau meej hauv Table 1, TPC cov ntsiab lus yog 67.49 mg GAE g-1. Cov ntsiab lus TC yog 3.51 µg g-1. Cov ntsiab lus TF thiab TL yog 49.78 thiab 17.45 mg QE g-1, feem. Ntxiv mus, DPPH-RSA thiab ABTS-RSA tau siv los ntsuas qhov kev nce qib ntawm cov haujlwm antioxidant. Cov txiaj ntsig tau qhia 128.71 µmol ntawm TE g-1 thiab 141.92 µmol ntawm TE g-1 rau DPPH-RSA thiab ABTS-RSA, raws li. Tsis tas li ntawd, lubkev ua haujlwm antioxidant(AOA) ntawm A. hierophantic yog nthuav tawm nyob rau hauv Table 1. Qhov feem pua ​​ntawm inhibition ntawm linoleic acid radicals raug xam raws li 45.74 feem pua ​​​​piv rau BHA siv -Carotene bleaching (-CB) assay. Tsis tas li ntawd, kev ntsuam xyuas ntawm cov hlau-chelating kev ua si tau qhia 42.89 mg g-1, uas zoo li muaj peev xwm cuam tshuam nrog Fe2 ntxiv -ferrozine complex tsim, qhia nws lub peev xwm los chelate oxidation hlau.

3.2. Quantification ntawm A. hierophantic Phenolic Compounds

Kev soj ntsuam kom muaj nuj nqis ntawm phenolic tebchaw rau KEE thiab KAE los ntawm HPLC kev tshuaj xyuas tau ua tiav, thiab cov ntaub ntawv tau teev nyob rau hauv Table 2. Cuaj qhov sib cais phenolic acids thiab rau flavonoids tau txheeb pom nyob rau hauv cov nyiaj tau los ntawm KEE ntawm A. hierophantic. Feem ntau cov phenolic acids yog hydroxycinnamic acids xws li sinapic acid (28.704 mg 100 g−1) ua raws li caffeic acid (6.621 mg 10{ {61}} g−1), rosmarinic acid (2.884 mg 100 g−1), ferulic acid (1.854 mg 100 g−1), thiab cinnamic acid (0.094 mg 100 g−1); thiab hydroxy-benzoic acids xws li p-hydroxybenzoic acid (3.440 mg 100 g-1), protocatechuic acid (1.811 mg 100 g-1), vanillic acid (3.326 mg 100 g-1), thiab syringic acid (1.083 mg 100 g. -1). Flavonoids xws li myricetin (16.269 mg 100 g-1), D-catechin (2.410 mg 100 g-1), kaempferol (0.434 mg 100 g-1), rutin (0.539 mg 100 g-1), apigenin {{56} }glucoside (0.192 mg 100 g-1), thiab quercetin (0.184 mg 100 g-1) raug kuaj pom tsis muaj nuj nqis. Cov phenolic tebchaw nyob rau hauv KAE ntawm A. hierophantic kuj tau txiav txim, thiab cov ntaub ntawv yog tabulated nyob rau hauv Table 2. Syringic acid tau sau tseg raws li lub siab tshaj plaws phenolic acid ntawm 21 phenolics txheeb xyuas. Catechol thiab pyrogallol yog 2.526 thiab 1.589 mg 100 g-1, raws li. Cov ntaub ntawv

qhia tias qee cov phenolic acids xws li caffeic, catechin, chlorogenic, epicatechin, e-vanillic, p-hydroxybenzoic, thiab protocatechuic acids tau kuaj pom nyob rau hauv qhov nruab nrab ntawm 0.725, 0.256, { {6}}.136, 0.193, {{10}}.443, 0.223, thiab 0.454 mg 100 g−1, feem. Nyob rau hauv tib lub ntsiab lus, tsawg npaum li cas ntawm 3,4,5-lub sij hawm ethoxy cinnamic, 4-aminobenzoic, benzoic, cinnamic, coumarin, ellagic, ferulic, gallic, iso-ferulic, -coumaric, p-coumaric , thiab salicylic acid tau txheeb xyuas tom qab raug txheeb xyuas. Epicatechin thiab D-catechin li flavonoids tau muaj nyob hauv KAE ntawm A. hierophantic ib yam nkaus.

3.3. Serum Creatinine, Urea, K, Tag Nrho Protein, thiab Albumin Qib

Kev txhaj tshuaj CCl4 tau txhawb nqa cov ntshav creatinine, urea, thiab k qib hauv GII nas thaum piv rau cov nas tswj (GI). Hloov pauv, tag nrho cov protein thiab albumin qib tau txo qis hauv CCl4-kho nas (Table 3). Vit. E ntxiv rau Se thiab A. hierophantic extracts (G III, IV, V, thiab VI) txo cov kev hloov pauv hauv creatinine thiab urea los ntawm kev txhaj tshuaj CCl4, thaum lawv nce albumin thiab tag nrho cov proteins kom ze rau cov txiaj ntsig ib txwm muaj hauv GI (Table 3. ). Serum k qib tau nce ntxiv hauv CCl4- kho nas (GII) thaum piv rau GI (Table 3). Kev txhaj tshuaj vit. E plus Se thiab kev tswj hwm ntawm A. hierophantic alcoholic thiab aqueous extracts (G IV, V, thiab VI) kuj tau txhim kho qib k thaum piv rau GI (Table 3).

3.4. Lub raum Antioxidant Biomarkers

Raws li qhia hauv Table 4, kev tswj hwm ntawm CCl4 txo qis SOD thiab GSH qib thiab nce qib MDA hauv GII.raumhomogenate cov ntaub so ntswg. Txawm li cas los xij, thaum piv rau GI, nas kho nrog ob qho tib si vit. E plus Se thiab A. hierophantic extracts (GIII, VI, V, thiab VI) tau nthuav tawm qhov kev txhim kho ntawm cov enzymes antioxidant SOD thiab GSH, nrog rau kev txo qis hauv MDA qib (Table 4). A. hierophantic alcoholic extract (GIV) outperformed A. hierophantic aqueous extract (GV) thiab ua ke A. hierophantic alcoholic thiab aqueous extracts nyob rau hauv attenuating antioxidant theem, thiab combating cov txheej txheem autoxidation ua rau cov MDA qis thaum piv rau GI.

3.5. Nephroprotection feem pua

Qhov feem pua ​​​​nephroprotection (kwv yees rau kev tswj tsis zoo (GI) thiab zoo (GII) pawg) ntawmraumkev ua haujlwm xws li creatinine, urea, k, TP, thiab albumin nrog rau kev ua haujlwm antioxidant hauv lub raum homogenate (MDA, SOD, GSH) yog qhia hauv Table 5. Cov nephroprotection feem pua ​​​​tau sau cov nqi siab tshaj li creatinine, urea, k hauv GIII, TP, thiab albumin hauv GV, MDA, thiab GSH hauv GIII thiab SOD hauv GV (Table 5). Tag nrho nephroprotection feem pua ​​txheeb ze rau vit. E plus Se kev kho mob tau sau npe siab tshaj plaws hauv pawg KAE kho (GV, 97.62 feem pua), tom qab ntawd KEE (GIV, 83.27 feem pua), thiab tom qab ntawd KEE ntxiv rau KAE (GVI, 78.85 feem pua), raws li qhia hauv Table 5.

3.6. Cov teebmeem ntawm A. hierophantic Extracts ntawm lub raum Histoarchitecture

Cov txiaj ntsig ntawm kev tshawb nrhiav biochemical tau txais kev txhawb nqa los ntawm kev kuaj histopathological. Table 6 thiab Daim duab 1 qhia txog qib ntawm cov kev hloov hauv histological hauv cov qauv hauv qab ntawm tus nas.lub raumnyob rau hauv ntau pawg sim kho nrog A. hierophantic extracts. Hauv kev tshawb nrhiav tam sim no, lubraumntawm pawg tswj hwm (GI) tau pom tias muaj cov qauv histological (Daim duab 1(I.1)). Lub histoarchitecture ntawm CCl 4- kho nas (GII) pom focal inflammatory cell infiltration (ntxiv rau ntxiv) ntawm cov tubules ntawm lub cortex, congestion (ntxiv rau ntxiv) ntawm cov hlab ntsha ntawm cov tubules (Daim duab 1(II.2)). ), ntau eosinophilic cam khwb cia (ntxiv rau ntxiv) tsim nyob rau hauv lub lumen ntawm ib co tubules, thiab focal hemorrhage (ntxiv rau ntxiv) ntawm cov degenerated tubules ntawm corticomedullary feem (Daim duab 1(II.3), Table 6). Hauv GIII, txhaj tshuaj vit. E plus Se, tswj cov dej cawv extract ntawm A. hierophantic (GIV), aqueous extract ntawm A. hierophantic (GV), thiab kev sib xyaw ntawm A. hierophantic extracts (GVI) attenuated cytotoxic teebmeem ntawm CCl4 thiab kaw me me (ntxiv) rau nruab nrab (ntxiv rau ntxiv) congestion nyob rau hauv cov hlab ntsha ntawm tubules ntawm lub cortex (Daim duab 1(III.4-VI.8), Table 6) nrog zoo-tsim Bowman's capsule nrog glomerulus thiab convoluted tubules enlarged. Tsis tas li ntawd, cov aqueous extract ntawm A. hierophantic (GV) tau sau tseg focal inflammatory cell infiltration (ntxiv rau ntxiv) ntawm corticomedullary feem (Daim duab 1(V.7), Table 6).

4. Kev sib tham

Phytochemicals feem ntau ua tau zoo dawb radical scavengers thiab suav hais tias yog cov nroj tsuag zoo tshaj plaws antioxidants. Cov tshuaj polyphenolic tau xav tias muaj cov tshuaj tiv thaiv kab mob thiab tiv thaiv mutagenic hauv tib neeg [40]. Ib qho tseem ceeb TPC cov ntsiab lus hauv A. hierophantic yog me ntsis siab dua qhov tau los ntawm Mohamed li al. [21] as 51.97 mg GAE g × 1 in A. hierophantic tshuaj ntsuab thiab los ntawm AlGamdi li al. [41], leej twg pom 4 mmol L × 1 GAE hauv A. hierophantic noob. Tsis ntev los no, Zin et al. [42] tau qhia txog qhov muaj tannins hauv A. hierophantic raws li bioactive compound thiab pom zoo nws bioactivity, uas yuav tsum tau tshawb xyuas tob. Cov ntsiab lus -carotene, uas yog ib feem ntawm tag nrho cov carotenoids, yog 2.27 µg g × 1 raws li tau hais los ntawm Mohamed li al. [21], thiab txawm tias tam sim no cov txiaj ntsig tau nthuav tawm tag nrho carotenoids li 3.51 µg g × 1 . Cov txiaj ntsig zoo sib xws hauv cov ntsiab lus flavonoid thiab flavonol tau qhia los ntawm Mohamed li al. [21]. Cov tshuaj lom neeg lom neeg, xws li phenolic compounds, tam sim nokev ua haujlwm antioxidantRaws li kev tawg ntawm lipid oxidation saw cov tshuaj tiv thaiv los ntawm kev muab hydrogen rau cov dawb radicals. Qhov kev tshem tawm cov peev txheej ntawm phenolics los tiv thaiv cov radicals yog elucidated los ntawm lawv cov phenolic hydroxyl pawg [8,10,22]. Cov phenolic acid no tau piav qhia tias yog cov tshuaj tiv thaiv antioxidant zoo, suav nrog hydrogen peroxide, hydroxyl radical, thiab superoxide anion [43]. A. hierophantic hlau chelating kev ua haujlwm zoo li ua tau zoo hauv kev cuam tshuam nrog "Fe2 plus -ferrozine" kev tsim kho, qhia nws lub peev xwm los ntes "ferrous" ions ua ntej "ferrozine". Kev sib raug zoo ntawm kev nce hauv lawv cov ntsiab lus ntawm phenolic tebchaw yog qhia ncaj qha nrog lawv cov peev xwm antioxidant [42]. Andjelkovi'c et al. [44] tsim cov kev ua ntawm ntau phenolic acids los tsim complexes nrog hlau. Qhov muaj txiaj ntsig TPC thiab cov haujlwm muaj txiaj ntsig antioxidant siv cov kev ntsuas sib txawv muab lub phaj meej los ntawm thiab lees paub qhov bioactivity ntawm A. hierophantic ua tshuaj ntsuab rau zaub mov lossis dej haus.

Cov tshuaj tiv thaiv antioxidant zoo, suav nrog hydrogen peroxide, hydroxyl radical, thiab superoxide anion [43,45,47]. Cov tshuaj uas tau txheeb xyuas thiab muaj nuj nqis los ntawm HPLC hauv KAE ntawm A. hierophantic tau siab dua li cov txheeb cais hauv KEE, tab sis txheeb xyuas cov tebchaw hauv KEE ntawm A. hierophantic tau nthuav tawm ntau dua [22]. Cov txiaj ntsig qhia tau hais tias kev siv A. hierophantic tuaj yeem nthuav qhia ntau yam hauv ob qho tib si polar thiab non-polar. Cov txiaj ntsig no zoo sib xws los ntawm AlGamdi li al. [41] raws li lawv tau txheeb xyuas thiab ntsuas 20 polyphenolic tebchaw hauv cov noob ntawm A. hierochuntica. Cov extract muaj chlorogenic acids thiab hydroxybenzoic acids, tab sis lub ntsiab Cheebtsam yog flavone C-glycosides, C-diglycosides, O-glycosides, thiab O-glycoside-C-glycosides tshwm sim feem ntau li luteolin conjugates. Tsis tas li ntawd, 14 ntawm 20 lub tebchaw hauv A. hierophantic extract tau nthuav tawmkev ua haujlwm antioxidantsiv HPLC-on-line tshuaj tiv thaiv kab mob antioxidant [41]. Interestingly, cov ntaub ntawv tam sim no tau lees paub tias A. hierophantic yog nplua nuj nyob rau hauv phytochemical tebchaw thiab yog ib qhov zoo ntawm tej yam ntuj tso antioxidants uas muaj peev xwm noj qab haus huv cov kev pab cuam, raws li tau tsis tshua muaj highlighted ua ntej nyob rau hauv cov noob [41]. Li no, tshuaj yej npaj los ntawm tag nrho cov nroj tsuag hmoov yog cov tsoos daim ntawv ntawm noj; cov ntaub ntawv qhia txog cov tshuaj bioactive tshuaj nyob rau hauv KEE thiab KAE ntawm A. hierochuntica, uas txawv ntawm cov uas pom nyob rau hauv AlGamdi li al. [41]

Hauv ntau cov kev tshawb fawb, CCl4-induced nephrotoxicity yog siv los ua qauv qauv rau kev kuaj cov nephroprotective nyhuv ntawm cov nroj tsuag rho tawm / tshuaj [48,49]. Txoj kev tshawb fawb tam sim no tau saib cov txiaj ntsig ntawm A. hierophantic extracts ntawm CCl4-inducedraumkev puas tsuaj, nrog rau nws cov nephroprotection thiab antioxidant muaj peev xwm nyob rau hauv nas. Hauv kev tshawb fawb tam sim no, pawg CCl4 kev kho mob (GII) tau nce qib creatinine, urea, thiab k thiab txo tag nrho cov protein thiab albumin concentrations thaum piv rau GI. Qhov no tej zaum yuav yog vim CCl4 intoxication yog ib qho tseem ceeb ntawm cov dawb radical tsim nyob rau hauv ntau yam kabmob, nrog rau lub siab, ob lub raum, ntsws, hlwb, thiab ntshav [50]. Nws kuj tau pom tias tom qab kev txhaj tshuaj CCl4 hauv cov nas, qhov concentration ntawm CCl4 tau faib ntau qhov sib npaug hauv ob lub raum tshaj hauv daim siab [51], txij li thaumraummuaj kev sib raug zoo siab rau CCl4 thiab muaj cytochrome P450, feem ntau hauv lub cortex. Feem ntau cov dawb radicals los ntawm CCl4 yog trichloromethyl radical (CCl3• ) thiab trichloromethyl peroxyl radical (CCl3O2• ) [52]. Cov radicals no txuas mus rau ib qho intracellular protein, cell membrane lipids, thiab DNA, ua rau cov protein denaturation, lipid peroxidation, thiab oxidative DNA puas tsuaj uas ua rau cell tuag [53]. Hauv qhov sib piv, kho CCl4- nas nrog vit. E plus Se (GIII) thiab A. hierophantic extracts (GVI: VI) tau txais txiaj ntsig zoo ntawm cov kev nce qib hauv creatinine thiab urea ntau ntxiv nrog rau cov ntshav albumin thiab tag nrho cov proteins kom ze rau lawv cov qib hauv GI. Qhov no tej zaum yuav yog vim muaj antioxidant zog thiab nplua nuj phenolic cov ntsiab lus ntawm A. hierophantic extracts thiab antioxidant muaj peev xwm thiab chelating kev ua ntawm vit. E plus Se, uas tshem tawm cov dawb radicals yog li inhibiting lub raum puas. Phytochemicals yog cov dawb radical scavengers uas muaj txiaj ntsig zoo tshaj plaws thiab suav tias yog cov tshuaj tua kab mob antioxidant zoo tshaj los ntawm cov nroj tsuag [54]. Feem ntau cov phenolic tebchaw yog hydroxycinnamic acids, xws li sinapic acid, ntawm cuaj phenolic tebchaw nyob rau hauv KEE, thaum syringic acid yog lub siab tshaj plaws phenolic acid ntawm 21 phenolic acids nyob rau hauv KAE. Rau flavonoids tau txheeb xyuas hauv KEE thiab ob qho hauv KAE siv HPLC kev tshuaj xyuas [55]. Tsis tas li ntawd, raws li antioxidant, vit. E ntseeg tias tiv thaiv cov ntaub so ntswg los ntawm kev puas tsuaj los ntawm cov pa oxygen metabolites. Selenium kuj tseem pom zoo tias yog ib qho tseem ceeb ntawm cov pob zeb hauv av rau tib neeg kev noj qab haus huv, tiv thaiv cov hlwb los ntawm kev puas tsuaj ntawm cov dawb radicals [22].

Hauv kev tshawb fawb tam sim no, CCl4 kev tswj hwm tau txo qis GSH thiab SOD thiab nce qib MDA hauvraumhomogenates txheeb ze rau GI. Vit. E plus Se thiab A. hierophantic extracts ameliorated ntau yam teebmeem ntawm CCl4 los ntawm kev kho cov kev hloov pauv ntawm cov tshuaj tua kab mob antioxidant xws li SOD thiab GSH thiab tuaj yeem cuam tshuam cov txheej txheem ntawm kev tsim MDA, raws li tau tshaj tawm tsis ntev los no [15,21,40,41] . GSH yog ib qho uas tsis yog-enzymatic antioxidant uas muaj nyob rau hauv tag nrho cov mammalian hlwb. Nrog rau nws daim ntawv oxidized, GSSG, GSH ua ib cofactor rau ntau detoxifying enzymes (GPx, GST, thiab lwm yam) tiv thaiv oxidative kev nyuaj siab thiab tswj cellular redox tshuav nyiaj li cas [47]. Qhov kev tshawb pom no yog ua raws cov Khan thiab Siddique [56] thiab Makni li al. [57], leej twg tau tshaj tawm tias CCl4 txo qis GSH qib hauv nas raum. Kev kho mob nrog vit. E plus Se thiab A. hierophantic extracts pom kev tiv thaiv kev txo qis hauv GSH qib tshwm sim los ntawm CCl4. Nyob rau hauv tib lub ntsiab lus, SOD catalyzes qhov dismutation ntawm ob lub molecules ntawm superoxide anion (*O2) rau hydrogen peroxide (H2O2) thiab molecular oxygen (O2), yog li ua rau muaj teeb meem superoxide anion tsawg txaus ntshai [58,59]. CCl4 intoxication hloov cov noob qhia theem los ntawm depleting raum SOD [60]. Kev txo qis hauv SOD kev ua haujlwm yog qhov ntsuas qhov ntsuas ntawm cov cellular puas. Peb kuaj A. hierophantic extracts ameliorated raum toxicity los ntawm alleviating qib ntawm SOD. Nws koom nrog ntau yam txheej txheem enzymatic los txo cov concentration ntawm cov tshuaj tiv thaiv detoxification [61]. MDA yog thawj cov khoom ntawm lipid peroxidation thiab yog ib qho ntawm cov cim tseem ceeb ntawm oxidative kev nyuaj siab. A. hierophantic extracts txo qis qhov nce hauv MDA qib thiab rov qab tag nrho lub zog antioxidant hauv CCl4- kho cov nas raum. Cov kev tiv thaiv no yuav yog vim muaj zog tiv thaiv kev ua haujlwm ntawm A. hierophantic extracts [15,21,40,41]. Cov txiaj ntsig no tseem qhia tau tias A. hierophantic extracts tuaj yeem ua rau txo qis oxidative kev nyuaj siab los ntawm kev txo qis lipid peroxide hauv CCl4-cov nas raum thiab tiv thaiv lub raum puas. Cov txiaj ntsig no tau pom zoo nrog cov txiaj ntsig ntawm kev ua haujlwm antioxidative ntawm Zn ntawm CCl4-induced mob nephrotoxicity [62,63].

A. hierochuntica extracts nthuav tawm muaj txiaj ntsig nephroprotection muaj peev xwm hais txograumKev ntsuas kev ua haujlwm (creatinine, urea, K, TP, thiab albumin) thiab lub raum homogenate antioxidant kev ua haujlwm (GSH, SOD, MDA) hauv GIV, V, thiab IV, feem. Tag nrho nephroprotection feem pua ​​txheeb ze rau vit. E plus Se kev kho mob tau sau npe qib siab tshaj plaws hauv KAE cov pab pawg kho mob (GV, 97.62 feem pua), tom qab ntawd KEE (GIV, 83.27 feem pua), thiab tom qab ntawd KEE ntxiv rau KAE (GVI, 78.85 feem pua), feem, raws li nqes los. Qhov no tej zaum yuav yog vim qhov sib txawv ntawm qhov ntau thiab qhov zoo ntawm phenolic thiab antioxidant cov ntsiab lus ntawm A. hierochuntica extracts, uas muaj kev cuam tshuam rau antioxidant muaj peev xwm [15,19,22,40,42].

Cov kev tshawb pom histopathological hauvlub raumyog ua raws li kev kwv yees biochemical ntawm pawg sim tshuaj ntsuam xyuas. CCl4 kev tswj hwm (GII) ua rau muaj qhov txhab glomerular thiab tubular lesion nrog vasocongestion hauv ob lub raum. Dogukan et al. [64] nrhiav pom tus qauv histological zoo sib xws hauv cov nas lub raum cov ntaub so ntswg hauv teb rau kev kho CCl4 ntev. Nws kuj tseem suav hais tias kev hloov pauv histological yog tshwm sim los ntawm kev ua haujlwm ntau dhau ntawm nephrons, uas ua rau lub raum tsis ua haujlwm [65], thiab / lossis yog vim kev puas tsuaj ntawm cov ntaub so ntswg provoked los ntawm kev tsim dawb radical ntawm CCl4 metabolism [56,66] . Cov nyhuv ntawm vit. E plus Se thiab A. hierophantic extract los kho thiab kho lub raum kev puas tsuaj ntawm CCl4 yog qhov tseem ceeb. Qhov no tej zaum yuav yog vim vit. E plus Se (raws li lub zog antioxidant) ua rau ROS ntxias los ntawm CCl4 [67]. A. hierophantic extracts suppress CCl4-induced mob nephrotoxicity vim lub luag hauj lwm antioxidative thiab dawb radical scavenging zog ntawm cov phenolic tebchaw muaj nyob rau hauv A. hierochuntica extracts [22]. Peb qhov kev tshawb pom tau zoo ib yam nrog rau lwm cov kws tshawb fawb uas tau pom tias ntau yam cog derivatives muaj cov tshuaj pharmacological los ntawm kev tshem tawm CCl4 kev tsim txom thiab rov ua kom zoo li qub [6].

5. Cov lus xaus

Cov txiaj ntsig ntawm txoj kev tshawb fawb no tau pom meej meej tias A. hierophantic cog yog nplua nuj nyob rau hauv polar thiab nonpolar phenolic tebchaw nrog lub peev xwm zoo tshaj plaws antioxidant, uas yog ncaj qha ntsig txog cov ntsiab lus phytochemical. A. hierophantic (tshwj xeeb yog aqueous extract) tiv thaiv nas tiv thaiv CCl4-induced oxidative stress thiab mobraumKev raug mob, raws li pov thawj los ntawm kev poob qis hauv MDA qib thiab nce GSH thiab SOD kev ua haujlwm, nrog rau kev txiav tawm ntawm biochemical thiab histological hloov pauv hauv lub raum. Kev tiv thaiv kev ua tau zoo tuaj yeem tshwm sim los ntawm cov antioxidant thiab dawb radical scavenging zog ntawm cov phenolic tebchaw uas muaj nyob rau hauv cov A. hierophantic extracts. Cov yam ntxwv no pab piav qhia txog cov nroj tsuag tshuaj zoo li tshuaj ntsuab. Kev tshawb fawb ntxiv yog xav tau los piav qhia tag nrho cov ntsiab lus tseem ceeb hauv A. hierophantic, thiab qhov kev tshawb fawb no yog tsim los txhawb kev tshawb fawb ntau ntxiv kom muaj cov ntaub ntawv txaus thiab cov lus pom zoo rau kev txheeb xyuas nws cov txheej txheem thiab kev noj tshuaj zoo.

Tus sau kev koom tes:Conceptualization, TIA, YMA thiab HB; methodology, kev tshawb nrhiav, HB thiab HAA-R.; data curation, TIA and YMA; kev sau ntawv-kev npaj ua ntej, tshuaj xyuas, thiab kho; HB and HAA-R. Txhua tus kws sau ntawv tau nyeem thiab pom zoo rau cov ntawv luam tawm ntawm cov ntawv sau.

Nyiaj txiag:Cov kws tshawb fawb xav ua tsaug rau Deanship of Scientific Research, Qassim University, rau kev pab nyiaj rau kev tshaj tawm ntawm txoj haujlwm no.

Cov ntaub ntawv muaj nyob:Cov ntaub ntawv nthuav tawm hauv qhov kev tshawb fawb no muaj nyob rau ntawm kev thov los ntawm tus kws sau ntawv.

AbbreviationsABTS:2,2'-azino-bis(3-ethylbenzene thiazoline-6-sulfonic acid); AOA:kev ua haujlwm antioxidant; BHA: butylated hydroxyanisole; BHA: butylated hydroxyanisole; DPPH: 1,1-diphenyl-2-picryl hydrazine; DW: qhuav hnyav; GA: gallic acid; GAE: Gallic acid sib npaug; GSH: txo-glutathione; HPLC-DAD: kev ua haujlwm siab ua kua chromatography diode array nrhiav pom; KAE: A. hierophantic aqueous extract; KEE: A. hierophantic ethanolic extract; MDA: malonaldehyde; QE: quercetin sib npaug; RAA: txheeb ze antioxidant kev ua haujlwm; ROS: reactive oxygen hom; RSA: radical scavenging kev ua si; Selenium; SE: txheem yuam kev; SOD: superoxide dismutase; TC: tag nrho carotenoids; TC: tag nrho carotenoids; TF: tag nrho cov flavonoids; TE: Trolox sib npaug; TFL: tag nrho cov flavonols; TPC: tag nrho cov phenolic compounds.

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