Urric acid Txo cov nyhuv thiab kev siv tshuab ntawm kev ua yeeb yam thiab echinacoside extracted los ntawm cistanche tubulosa
Mar 03, 2025
Abstract:Tsom mus tshawb cov uric acid under cov teebmeem thiab cov txheej txheem ntawm kev ua yeeb yam (li) thiab Ech) rho tawm los ntawmCistanche tubulosa.
TxojkevHyperuricemia (Hua) tus qauv muaj teeb tsa siv cov poov tshuaj oxonate (1.5 G1}}}}}}}}}}}}}}). Hua tus qauv nas tau muab faib ua cov qauv tswj hwm ua qauv, muaj cov pab pawg al1}}}}}}}} Mg {{{16} {17}}}}) cov pab pawg ib pawg. Ib pawg neeg tswj ib txwm tau ua dhau los. Tom qab plaub lub lim tiam ntawm kev tswj hwm, cov ntshav biochemistry analyzer tau siv los tshawb fawb urea, crea, adenosine deaminase (ADA) thiab ura acid (Uua) qib hauv cov zis. Enzyme txuas immunosorbent assay tau siv los kuaj Xanthine oxidase (xod) qib hauv ntshav dej. Raum tau sau, thiab lub raum coefficient tau hnyav thiab xam. Hematoxylin Eosin (nws) Staining tau siv los saib cov kev hloov pauv ntawm lub raum. Immunohistochemistry (IHC) thiab lub sijhawm pcr tau siv los tshawb xyuas cov protein thiab mrat1), organic anion Transporter 3 (OAT3) nyob rau hauv cov nqaij daim tawv. Cov txiaj ntsig tau raws li thiab Ech tuaj yeem txhim kho cov xim muaj zog ntawm ADA thiab XRAM
Cov lus xaus rau phenylethanoid glycosides raws li thiab Ech los ntawm cistal muaj nuj nqi los ntawm kev nthuav qhia cov kev qhia ntawm URAT1, glut9, oat1, thiab oat3.
Ntsiab lus: Cistance tubulosa; ua yeeb yam; Echinacoside; hyperuricemia; txo cov kua qaub; lub tshuab
Cistanche tubulosa extract nrog 16% ua yeeb yam
Hyperuricemia (HUA) yog tus mob metabolic tau los ntawm kev tsis taus ntawm tib neeg lub cev, ua rau cov ntshav nce siab uric acid acid qib [1]. Nws yog feem ntau kuaj pom thaum cov ntshav}}}}} {}}} μd} μmolocliv (357} μd} μd} μd} μmol.c. Cov kev txhim kho ntawm cov neeg hyperuricemia yog feem ntau xws li kev haus zaub mov huv, siv dej cawv ntev, thiab siv tshuaj hypothyroidism. Nws tsis tsuas yog ua kom muaj kev pheej hmoo ntawm cov kab mob plawv, ntshav qab zib, thiab kub siab [}] tab sis kuj cuam tshuam ntau txoj kev noj qab haus huv ntawm cov neeg mob.
Tam sim no, kev kho mob kho mob rau cov tshuaj phem uas cuam tshuam nrog cov tshuaj uas cuam tshuam thiab txhawb rictric accretion. Txawm li cas los xij, cov kev kho no muaj qee qhov kev txwv rau cov neeg mob lub siab lossis raum ua haujlwm [{{}]. Yog li no, tshawb txog cov tshuaj zoo nrog cov kev mob tshwm sim qis yog qhov tseem ceeb rau kev tiv thaiv thiab kho cov hyperuricemia.
Cistanche tubulosa (Schenk) R. Wight, feem ntau yog siv tshuaj suav tshuaj ib txwm muaj tshuaj ntsuab, yog ua haujlwm rau cov neeg mob uas tsis muaj peev xwm, kev ua txhaum cai, thiab cem quav vim mob plab hnyuv [7]. Cov kev tshawb fawb tau pom tiasCistanche tubulosamuaj cov neuroprotrective [8], anti-inflammatory [10], thiab kev ua tau zoo-txhim kho cov tshuaj kho mob rau cov neeg mob cancer [11]. Ntawm nws ntau ntau yam kev ua hauj lwm, phenylethanoid glycosides yog qhov muaj ntau tshaj plaws.
Actionoside (AS) thiab E NECHACOSIDE (E NECH), ob chav phenylethanoid glycosides nrog cov ntsiab lus siab hauvCistanche tubulosa, nthuav qhia muaj cov kev ua haujlwm muaj txiaj ntsig muaj txiaj ntsig zoo thiab tuav lub peev xwm tseem ceeb rau kev txhim kho thiab kev thov. Thiaj li, peb pawg tshawb fawb raug xaiv raws li thiab Ech rau kev kawm ntxiv. Los ntawm network portortry kev twv ua ntej thiab kev sim ua ntej hauv nas, peb pom tias Phenylethanoid glycoside extractions los ntawmCistanche tubulosaTej zaum yuav muaj qee yam urric acid-butming nyhuv. Txawm li cas los xij, txoj hauv kev thiab cov txheej txheem ntawm cov teebmeem no tseem tsis paub meej.
Yog li, siv cov txheej txheem ntawm cov kua qaub inewion thiab nws cov kev tshawb fawb ua ntej, peb tshawb xyuas cov txiaj ntsig enylethanoid glycoside muab rho tawm raws li thiab Ech. Qhov kev kawm no lub hom phiaj rau kev tshem tawm txoj hauv kev thiab cov txheej txheem los ntawm uas ua thiab Ech ua rau lawv cov txiaj ntsig ecour-txo qis.
1 Khoom
1.1 Cov tsiaj sim
Xya Caum Cuav Sprague-DD) nas (SD) nas, uas muaj los ntawm kev sim tsiaj ntawm Xinjiang Medical University of Xinjiang Medical University. Daim Ntawv Tso Cai Ntau Yam Tsiaj: Scxk (Xin) {4 4}}. Qhov kev sim no tau pom zoo los ntawm Pawg Neeg Txom Nyem ntawm Xinjiang Autonomous cheeb tsam Uygur Kev Pom Zoo Tshuaj Pom Zoo Tus lej: IMUN) {6}}.
1.2 tshuaj thiab reagents
Phenylethanoid glycoside muab rho tawm (raws li Ech)ntawmCistanche tubulosa (purity >98%, txheeb xyuas qhov kev ua tau zoo chromoography) tau muab rho tawm, huv si los ntawm lub chaw tshawb fawb txog kev tshawb fawb txog kev tshawb fawb txog. Cov lej lej: 2022017 (li) thiab 20220526 (E NECH).
Allopurinol ntsiav tshuaj, tus lej batch: 62208726, manufacture Jiangsu Wanbang Biochemical Group Co., Ltd.
Poov tshuaj oxonatethiabAdenine, tus lej Batch: I2130050 thiab C2202255, yuav los ntawm Shanghai Aladdin Biochemical Technology Co., Ltd.
Crea (Creatinine), UA (uric acid), urea, thiab ada (adenosine deaminase) assay cov khoom siv, tus lej Batch: 141123010, 14122004, 1413008, thiab 043624005, yuav los ntawm Shenzhen Minday Bio-Medical Electronics Bio, Ltd.
Xod (Xanthine Oxidase) Ua Haujlwm Assay Khoom, tus lej batch: y10001219, yuav los ntawm Shanghai Macklin Biochemical Technology Co., Ltd.
Cov tshuaj tiv thaiv (URAT1, glut9, oat1, oat3), cov lej lej (3i5723 0, 92F0296 / 4, tau yuav los ntawm kev muaj affinity biosciences.
HRP-daim ntawv lo rau tshis tawm tsam-luav IGG, tus lej batch: 245440113, muas ntawm Zsgb-Bio.
Hematoxylin Staining daws, tus lej batch: 24135741h, yuav los ntawm Beijing Lanjieke Technology Co., Ltd.
Dab Neeg Chromogenic Reagent Cov Khoom Siv, tus lej batch: 240010512, yuav los ntawm zsgb-bio.
Depc (Daivethyl pyrocarbonate), Tus lej catalog: D {60029-500 ml, yuav los ntawm Amrosesco (USA).
M5 Hiclear DL2000 DNA Cim, Tus lej catalog: MF025, yuav los ntawm Beijing Genomics Biotechnology Co., Ltd.
Trizol ™ Reagent, Tus lej catalog: 15596026, yuav los ntawm Thermo Fisher Sciety (USA).
5x tag nrho-hauv-ib rt mastermixthiabEvagreen qhia 2 × QPCL Mastermix-Tsawg Rox, catalog tooj: G492 thiab G891, ob qho tib si yuav los ntawm kev ua ntawv thov cov ntaub ntawv biological Inc. (ABM).
2 txoj kev
2.1 qauv tsim thiab pawg
Xya Caum Txiv neej Sprague-DD) nas muaj acclimated thiab muab faib ua ob pawg neeg: ib qho kev tswj hwm ib txwm) thiab cov qauv tswj hwm (60 nas). Ib pab pawg tswj ib txwm tau muab 0.5% CMC-NA ntawm gavage. Tus qauv tsim qauv tau muab cov tshuaj pleev cov poov xab oxonate (1.5 GGLE) ua ke nrog Adenine (100 Mgthina) ntawm qhov koob tshuaj 10 MLACH, ib hnub ib hnub rau 14 hnub sib law liag. Tus qauv tau txiav txim siab tau ua tiav tau tsim thaum urea, crea, thiab cov qib sa nce ntxiv, raws li ntsuas los ntawm cov kws tshuaj ntsuam xyuas biochemical.
Cov qhab nia zoo nkauj nas tau muab faib ntxiv rau hauv cov pab pawg hauv qab no, cov pab pawg} Cov pawg} Ech Pab Pawg (100 Mg · Kg⁻¹), nrog 10 nas nyob hauv txhua pab pawg. Txhawm rau tswj cov hyperuricemia ruaj khov, txhua pab pawg tsuas yog siv cov poov tshuaj oxonate (500 mg · Kg-agonate (500) ntawm cov tshuaj nyob rau hauv pab pawg hauv pab pawg thaum tav su. Cov pawg neeg tswj kav ib txwm tau txais 0.5% CMC-na kev tov khoom nyob rau yav sawv ntxov thiab haus dej thaum tav su 10 mlay, ib hnub ib hnub rau 14 hnub sib law liag. Tom qab, kev tswj hwm ntawm cov qauv ua qauv raug nres, thiab cov tshuaj tau ua raws li txhua hnub rau 14 hnub ntxiv.
2.2 Qauv Sau
Tom qab cov koob tshuaj kawg, cov nas tau yoo mov thaum hmo ntuj (dej tsis txwv). Cov zis los ntawm txhua pab pawg tau sau rau uric kev sim tshuaj xyuas. Lub cev qhov hnyav tau ntsuas tom qab kev yoo mov, thiab kuaj ntshav tau sau hauv qab tshuaj loog. Ntshav dej tau sib cais los ntawm centrifugation thiab khaws cia rau kev siv yav tom ntej. Lub raum nqaij tau sau thiab luj. Ib lub raum tau tsau nyob rau hauv 4% pararormaldehyde tov, thaum lwm tus yog snap-khov hauv cov hmoov ua kua rau ntxiv kev tshuaj xyuas.
2.3 Kev ntsuas ntawm cov kua qaub hauv nas
Cov zis ua piv txwv los ntawm txhua pawg tau centrifuged (3, {{} {1} · Min⁻¹, 10 min, centrifuge lub vojvoog 14 cm), thiab supernatant tau sau. Cov zis ua piv txwv yog diluted 1: 9 nrog saline thiab txheeb xyuas rau uricid cov ntsiab lus siv cov kws tshaj lij biochemical.
2.4 Ntsuas ntawm Ada, urea, crea, thiab sua hauv ra dej hiav txwv
Cov ntshav piv txwv los ntawm txhua pawg tau centrifuged (3, {{}} · min⁻¹, 10 min, centrifuge xob luam 14 cm), thiab ntshav tau sau. Ada, urea, Irea, thiab cov theem SUA hauv cov ntshav dej tau ntsuas siv cov kws ntsuas cov neeg txheeb xyuas cov neeg siv tshuaj tua kab mob biochemical.
2.5 kev ntsuas ntawm Xod hauv nas Ntshav
Kev hloov pauv ntawm cov xod theem hauv cov kab mob nas tau ntsuas siv cov lus qhia enzymatic assay raws li cov chaw tsim khoom cov lus qhia. Qhov xub thawj nqus (AO) thiab kawg nqus tau (AO) ntawm 290 nm tau sau cia, thiab ΔA {1} {1} {1}) tau muab xam. ΔA tau hloov chaw rau cov qauv:
Kaum-· · txiv neej) {{{{{{0} × × × opt opt ÷ ÷ optical vease ÷ ÷}} × δa,
Txhawm rau laij cov nqi ntawm uric acid catalyzed ib feeb rau ib lub milliliter ntawm ntshav dej.
2.6 raum coefficient ntsuas
Lub raum nqaij tau sau, ntxuav nrog saline, thiab dej ntau heev tau raug tshem tawm. Lub cev muaj zog tau ntsuas, thiab lub raum coefficient raug suav raws li nram no:
Raum coefficient=Raum nyhav (mg) / lub cev hnyav (G).
2.7 histopathical kev tshuaj xyuas ntawm lub raum nqaij los ntawm nws staining
Lub raum nqaij ntswg ruaj ruaj nyob rau hauv 4% kev daws teeb meem pararorm yog lub cev qhuav dej nrog cov etaffrated ethale, thiab seem rau {} μm slices. Cov ntu yog deparafficized nrog xyab, hydrated nrog cov eteped ethanol, stained nrog Hematoxylin thiab EOSIN (nws), thiab muab kaw nrog nruab nrab ntawm cov pos hniav. Kev hloov kho morphological hauv lub raum nqaij tau pom nyob rau hauv lub qhov tsom tsom, thiab cov duab tau ntes ntawm 400 × magnification.
2.8 Nrhiav tau ntawm Urat1, glut9, oat1, thiab oat3 protein qhia hauv lub raum nqaij los ntawm IHC
Lub raum nqaij ntswg tsau hauv 4% kev daws teeb meem paraformaldehyde yog lub cev qhuav dej nrog cov etelhanol qhov ntau, tshem nrog cov kab mob paraffin, thiab seem. Antigen retiDieval thiab tshis cov ntshav thaiv tau ua. Thawj tshuaj tua kab mob (Urat1, glut9, OAT3, OAT3) tau ntxiv nyob rau ntawm 1: 200 Dilution thiab muab kom dhau ib hmos ntawm 4 degree. Hnub tom qab, cov ntu raug ntxuav nrog PBS, cua-qhuav, thiab tsim kom muaj pa nrog cov kab mob tshis ntawm 37 degree rau 30 feeb. Cov seem tau ntxuav nrog kab mob PBS thiab muab kaw rau hauv pbs chromogenic seeding, lithboric acids rau 3 feeb, ua kom muaj kev phom sij, thiab xhleej, thiab kev sib khi nrog cov hniav nruab nrab.
Rau txhua tus nas, ob random 4 {{{{0 0 × ib Nplooj 0 × cov teb ntawm lub raum ntu tau xaiv. Zoo staining (xim av-daj) tau txheeb xyuas qhov tseeb, thiab cov khoom siv kho tshiab (iod) ntawm txhua daim duab tau ntsuas siv 6.0 rau kev txheeb xyuas cov duab 6.0 rau kev txheeb xyuas cov duab 6.0 rau kev tsom xam.
