Dual Modulatory Effects Ntawm Diosmin Ntawm Calcium Oxalate Raum Pob Zeb Tsim Cov Txheej Txheem: Crystallization, Kev Loj Hlob, Kev sib sau ua ke, Crystal-cell Adhesion, Internalization rau hauv lub raum Tubular Cells, thiab ntxeem tau los ntawm Extracellular Matrix
Mar 16, 2022
Yog xav paub ntxiv:ali.ma@wecistanche.com
Supaporn Khamchuna,b,c,1, Sunisa Yoodeea,1, Visith Thongboonkerda,*
aMedical Proteomics Unit, Office for Research and Development, Kws qhia ntawv ntawm Medicine Siriraj Hospital, Mahidol University, Bangkok 10700, Thailand
bDepartment of Medical Technology, Tsev Kawm Ntawv ntawm Allied Health Sciences, University of Phayao, Phayao 56000, Thaib teb
cUnit of Excellence in Integrative Molecular Biomedicine, School of Allied Health Sciences, University of Phayao, Phaj 56000, Thailand
Keywords: Bioactive compound, Flavonoid, Inhibitor, Modulator, Nephrolithiasis, Txhawb nqa, Urolithiasis
TSAB NTAWV
Diosminyog lub ntuj flavone glycoside (bioflavonoid) pom nyob rau hauv cov txiv hmab txiv ntoo thiab cov nroj tsuag nrog ntau yam kev ua ub no. Nws tau dav siv los ua kev noj zaub mov ntxiv lossis kho tus neeg sawv cev hauv ntau yam kab mob / kab mob. Txawm hais tias pom zoo, pov thawj ntawm nws txoj kev tiv thaiv kev tiv thaivraumCov kab mob pob zeb (nephrolithiasis / urolithiasis), tshwj xeeb tshaj yog calcium oxalate (CaOx) monohydrate (COM) uas yog hom ntau tshaj plaws, tseem tsis paub meej. Nyob rau hauv txoj kev tshawb no, peb li systematically ntsuam xyuas cov teebmeem ntawmdiosmin(ntawm 2.5–160 nM) ntawm ntau theem ntawmraumCov txheej txheem tsim pob zeb, suav nrog COM crystallization, siv lead ua kev loj hlob, sib sau ua ke, siv lead ua-cell adhesion, internalization rau hauv lub raum tubular hlwb thiab ntxeem tau los ntawm extracellular matrix (ECM). Cov txiaj ntsig tau pom tiasdiosminmuaj koob tshuaj-dependent modulatory teebmeem rau tag nrho cov hais txog COM raum pob zeb txheej txheem.Diosminnce COM siv lead ua naj npawb thiab loj thaum lub sij hawm crystallization, tab sis txo cov siv lead ua loj thiab loj hlob. Thaumdiosmintxhawb nqa crystal aggregation, nws inhibited crystal-cell adhesion thiab internalization rau hauv lub raum tubular hlwb. Thaum kawg, diosmin txhawb nqa crystal ntxeem tau los ntawm ECM. Peb cov ntaub ntawv muab pov thawj qhia pom ob qho tib si inhibiting thiab txhawb kev cuam tshuam ntawm diosmin ntawm COMraumcov txheej txheem tsim pob zeb. Raws li cov kev ua ub no dual modulatory ntawm diosmin, nws lub luag hauj lwm los tiv thaiv urolithiasis yog tsis ntseeg thiab ceev faj yuav tsum tau ua rau nws siv nyob rau hauv.raumkab mob pob zeb.
1. Taw qhia
Diosmin(3′,5,7-trihydroxy-4′-methoxyflavone-7-rhamnoglucoside) yog lub ntuj bioflavonoid feem ntau pom muaj nyob hauv ntau yam nroj tsuag thiab txiv hmab txiv ntoo, feem ntau Citrus spp. [1,2]. Nws tuaj yeem raug tsim los yog muab los ntawm lwm cov flavonoid, hesperidin [1,2]. Diosmin ib leeg los yog ua ke nrog hesperidin yog dav siv los ua phlebotropic tshuaj rau kev kho mob ntawm venous thiab lymphatic mob xws li mob venous insufficiency thiab hemorrhoids [3,4]. Tsis tas li ntawd, diosmin nthuav tawm ntau lwm yam tshuaj lom neeg thiab tshuaj lom neeg, suav nrog kev ua haujlwm antioxidant thiab tiv thaiv kab mob [5,6], tshuaj tiv thaiv mutagenic thiab anti-neoplastic zog [7,8], tshuaj tua kab mob [9], cov khoom tiv thaiv hyperglycemic [ 10], thiab kev tiv thaiv los tiv thaiv ntau lub cev puas tsuaj, piv txwv li,
mob plawv [11], retinal [12],raum, daim siab thiab lub hlwb raug mob [13].
Lub raumpob zeb kab mob (los yog nephrolithiasis / urolithiasis) yog tshwm sim los ntawm cov khoom calculi tsim thiab tso rau hauv lub raum thiab cuam tshuam rau tib neeg nyob rau hauv tag nrho cov cheeb tsam ntawm lub ntiaj teb no nrog ib tug siab recurrence rate [14-17]. Ntawm tag nrho cov khoom siv lead ua ntau yam, calcium oxalate (CaOx), tshwj xeeb tshaj yog monohydrate daim ntawv (COM), yog cov kab mob feem ntau thiab cov crystalline feem ntau pom nyob rau hauv cov pob zeb qub (cov neeg mob raum pob zeb) [18]. Mechanistically, COM crystals muaj lub peev xwm tshaj nplaum nplaum thiab feem ntau cytotoxic cuam tshuam rau lub raum tubular epithelial hlwb [19,20]. Thaum lub sij hawm lub pob zeb pathogenesis, ob qho tib si Randall cov quav hniav qauv thiab intratubular hypothesis muaj cov yam ntxwv ntawm COM pob zeb tsim txheej txheem, nrog rau COM crystallization, kev loj hlob, sib sau ua ke, khaws cia los ntawm qhov chaw adhesion ntawm lub raum tubular hlwb los yog lub raum pelvis, internalization rau hauv lub hlwb, thiab ntxeem tau rau hauv. raum interstitium los ntawm extracellular matrix (ECM) [21,22].
Tsis ntev los no, ntau cov kev tshawb fawb tau tsom mus rau kev tshawb pom cov tshuaj siv cov nroj tsuag / txiv hmab txiv ntoo thiab lawv cov bioactive compounds tsom rau kev tiv thaiv cov pob zeb tshiab thiab / lossis rov tshwm sim. Ib tug xov tooj ntawm cov ntaub ntawv yav dhau los tau qhia tias ib co bioactive tebchaw, tshwj xeeb tshaj yog phenolic tebchaw (polyphenol, flavonoid, flavone glycoside, thiab lwm yam) muab rho tawm los ntawm ntau yam tshuaj nroj tsuag / txiv hmab txiv ntoo, muaj kev tiv thaiv los ntawm cov pathogenic mechanisms ntawm cov tshuaj.raumkab mob pob zeb ob leeg hauv vitro thiab hauv vivo [23–25]. Ntawm cov khoom muaj txiaj ntsig zoo no, ob peb tsab ntawv ceeb toom tau qhia tias diosmin tuaj yeem tiv thaiv kev tso tawm ntawm CaOx crystals hauv lub raum cov ntaub so ntswg hauv cov qauv tsiaj [26,27]. Txawm li cas los xij, nws lub luag haujlwm meej modulatory (inhibition lossis nce qib) thiab cov txheej txheem (theem lossis txheej txheem pob zeb) hauv COM lub raum pob zeb tsim tsis tau tshawb xyuas. Txoj kev tshawb no yog li systematically ntsuam xyuas cov teebmeem ntawmdiosmin(ntawm 2.5–160 nM) ntawm ntau theem ntawmraumCov txheej txheem tsim pob zeb, suav nrog crystallization, siv lead ua kev loj hlob, sib sau ua ke, siv lead ua-cell adhesion, internalization rau hauv lub raum tubular hlwb thiab ntxeem tau los ntawm ECM.
Nyem rauCistanche tubulosa ntau npaum li cas rau lub raum ua haujlwm
2. Cov ntaub ntawv thiab cov txheej txheem
2.1. Kev npaj Diosmin
Diosmin(Tokyo Chemical Industry; Tokyo, Nyiv) tau yaj nrog 100 feem pua dimethyl sulfoxide (DMSO) (Sigma-Aldrich; St. Louis, MO) thiab ua haujlwm aliquots tau ua nyob rau hauv qhov kawg concentrations ntawm 2.5, 5, 10, 20, 40, 80 thiab 160 nM rau kev tshawb xyuas nws cov teebmeem ntawm COM crystals. Tsis tas li ntawd, 100% DMSO tau siv los ua qhov kev tswj tsis zoo hauv txhua qhov kev sim.
2.2. Cell kab lis kev cai
MDCK lub raum tubular epithelial cell kab tau muab los ntawm distal tubular ntu ntawm nephron (ATCC; Manassas, VA) tau nthuav tawm hauv Eagle qhov tsawg kawg nkaus qhov nruab nrab (MEM) (Gibco; Grand Island, NY) ntxiv nrog 10 feem pua fetal bovine serum (FBS), 60 U/ml penicillin G (Sigma-Aldrich) thiab 60 ug/ml streptomycin (Sigma-Aldrich). Cov hlwb tau khaws cia rau hauv lub tshuab nqus tsev vaum nrog 5 feem pua CO2 ntawm 37 ◦C.
2.3. COM crystallization kev soj ntsuam
COM crystallization tau ua raws li tau piav ua ntej [28,29]. Luv luv, 500 ul ntawm 10 mM CaCl2⋅2H2O nyob rau hauv crystallization buffer (muaj 10 mM Tris-HCl thiab 90 mM NaCl) (pH 7.4) tau ntxiv rau hauv txhua qhov dej ntawm 24-zoo phaj (Corning Inc.; Corning, NY). Qhov sib npaug ntawm qhov ntim (4 ul) ntawm crystallization buffer (kev tswj tsis pub dawb), DMSO (kev tswj tsis zoo) lossisdiosmin(2.5, 5, 10, 20, 40, 80 lossis 160 nM) tau tov nrog CaCl2. Thaum kawg, 500 ul ntawm 1.0 mM Na2C2O4 nyob rau hauv lub crystallization tsis yog ces ntxiv mus ua qhov kawg concentrations ntawm CaCl2 thiab Na2C2O4 mus rau 5 mM thiab 0.5 mM, feem. Cov sib tov yog incubated ntawm 25 ◦C rau 1 h thiab ces kuaj. Cov duab siv lead ua tau raug ntes los ntawm yam tsawg kawg 15 qhov chaw siab zog (HPFs) nyob rau hauv Nikon Eclipse Ti-S inverted theem-contrast lub teeb microscope (Nikon; Tokyo, Nyiv). Crystal loj thiab tus lej raug ntsuas los ntawm tsawg kawg 15 HPFs rau txhua tus qauv siv NIS Element D software version 4.11 (Nikon). Crystal mass yog xam los ntawm tsawg kawg yog 100 crystals hauv 15 HPFs siv cov qauv hauv qab no: Crystal mass (µm2/HPF)=Nruab nrab siv lead ua loj hauv txhua daim teb (µm2) × Tus naj npawb ntawm cov muaju hauv txhua daim teb (/HPF) ( 1)
2.4. COM crystal kev loj hlob assay
Kev ntsuam xyuas Crystal loj hlob tau ua raws li tau piav qhia yav dhau los [30,31].
Luv luv, qhov sib npaug ntim (500 ul) ntawm 10 mM CaCl2⋅2H2O thiab 1.0 mM Na2C2O4 hauv crystallization tsis yog sib xyaw (1: 1) (v/v) hauv txhua qhov dej ntawm { {12}}zoo phaj. Qhov sib tov yog incubated ntawm 25 ◦C rau 1 h kom ua tiav crystallization. Ntawm qhov taw tes no (T0), qhov sib npaug ntawm qhov ntim (4 ul) ntawm crystallization buffer (kev tswj tsis pub dawb), DMSO (kev tswj tsis zoo) lossisdiosmin(2.5, 5, 1{{10}}, 20, 40, 80 lossis 160 nM) tau ntxiv rau hauv txhua qhov dej thiab qhov sib tov tau ntxiv rau 60 min (T60). Ntawm T0 thiab T60, cov duab siv lead ua tau raug ntes los ntawm yam tsawg kawg 15 HPFs nyob rau hauv Nikon Eclipse Ti-S inverted theem-contrast lub teeb microscope. Crystal qhov ntau thiab tsawg ntawm T0 thiab T60 tau ntsuas siv NIS Element D software version 4.11 (Nikon), qhov loj hlob siv lead ua (xws li Δ Crystal loj) tau suav los ntawm tsawg kawg 100 crystals hauv 15 HPFs siv cov qauv hauv qab no: Δ Crystal loj (µm2)=Crystal loj ntawm T60 - Crystal loj ntawm T0 (2)
2.5. COM crystal aggregation assay
Crystal aggregation assay tau ua raws li tau piav qhia yav dhau los [32,33]. COM crystals tau tsim raws li tau hais los saum toj no hauv kev ntsuas kev loj hlob siv lead ua tab sis nrog lub ntim loj dua hauv 50-ml conical raj (Corning Inc.) thiab tom qab ntawd sau los ntawm centrifugation ntawm 2000 g rau 5 min. Cov supernatant raug muab pov tseg, hos COM crystals tau ntxuav peb zaug nrog methanol. Tom qab lwm qhov centrifugation ntawm 2000 g rau 5 min, methanol tau muab pov tseg thiab cov crystals tau qhuav ib hmos ntawm 25 ◦C. COM crystals (1000 µg qhuav qhov hnyav) tau muab tshem tawm hauv 1 ml ntawm crystallization tsis nyob hauv txhua qhov dej ntawm 6-zoo phaj (Corning Inc.). Qhov sib npaug ntawm qhov ntim (4 ul) ntawm crystallization buffer (kev tswj tsis pub dawb), DMSO (kev tswj tsis zoo) lossisdiosmin(2.5, 5, 10, 20, 40, 80 lossis 160 nM) tau ntxiv rau hauv COM crystal suspension hauv txhua qhov dej. Lub phaj tau txuas ntxiv shaken nyob rau hauv lub co incubator (Zhicheng; Shanghai, Tuam Tshoj) ntawm 150 rpm thiab 25 ◦C rau 1 h. Tom qab ntawd, kev tsim ntawm COM siv lead ua aggregate (txhais tau tias yog "kev sib dhos ntawm peb lossis ntau dua ib tus neeg COM crystals uas nruj nreem koom ua ke" [32]) tau tshuaj xyuas thiab ua duab nyob rau hauv Nikon Eclipse Ti-S inverted phase-contrast light microscope. Tus naj npawb ntawm COM crystal aggregates raug suav los ntawm tsawg kawg 15 random HPFs ib qhov dej.
2.6. COM crystal-cell adhesion assay
Crystal-cell adhesion assay tau ua raws li tau piav qhia yav dhau los [34,35]. Luv luv, COM crystals tau tsim raws li tau hais los saum toj no hauv 50-ml conical raj thiab tom qab ntawd sau los ntawm centrifugation ntawm 2000 g rau 5 min. Cov supernatant raug muab pov tseg, hos COM crystals tau ntxuav peb zaug nrog methanol. Tom qab lwm qhov centrifugation ntawm 2000 g rau 5 min, methanol tau muab pov tseg thiab cov crystals tau qhuav ib hmos ntawm 25 ◦C. Cov crystals tau decontaminated los ntawm UV lub teeb hluav taws xob rau 30 feeb ua ntej kev cuam tshuam nrog cov hlwb.
MDCK hlwb (ntawm qhov ntom ntawm 2 × 105 hlwb / qhov zoo) tau noob thiab loj hlob nyob rau hauv txhua lub qhov dej ntawm 6-zoo phaj (Corning Inc.) rau 48 teev kom tau txais cov monolayer sib xyaw. Cov kab lis kev cai nruab nrab tau rov kho dua ua ntej ntxiv COM crystals (100 µg qhuav hnyav crystals ib ml kab lis kev cai nruab nrab). Qhov sib npaug ntawm qhov ntim (4 ul) ntawm crystallization buffer (kev tswj tsis pub dawb), DMSO (kev tswj tsis zoo) lossisdiosmin(2.5, 5, 10, 20, 40, 80 lossis 160 nM) tau ntxiv rau hauv txhua qhov dej. Cov hlwb tau muab txuas ntxiv rau hauv qhov chaw tsim khoom humidified nrog 5 feem pua CO2 ntawm 37 ◦C rau 1 h. Tom qab ntawd, cov hlwb raug ntxuav nrog PBS tsib zaug thiab ua duab nyob rau hauv Nikon Eclipse Ti-S inverted phase-contrast light microscope. Tus naj npawb ntawm cov seem COM crystals adhered mus rau lub raum tubular cell nto yog suav los ntawm tsawg kawg yog 15 random teb rau ib qhov dej.
Cistanche rau lub raum ua haujlwm
2.7. COM crystal internalization assay
Cov fluorescence-labeled COM crystals tau tsim raws li tau piav ua ntej [36,37]. Cov khoom sib xyaw / kev sib xyaw ua ke ntawm cov tshuaj siv rau crystallization yog tib yam li tau hais los saum toj no rau cov dawb (non-labeled) crystals, tab sis 0.1 µg/ml fluorescein isothiocyanate (FITC) (Thermo Scientific Pierce; Rockford, IL) yog ntxiv rau CaCl2⋅2H2O tshuaj ua ntej sib tov nrog Na2C2O4. Cov kauj ruam tom ntej (crystallization thiab sau) tau ua raws li tau hais los saum no, tab sis nyob rau hauv qhov tsaus ntuj.
Txhawm rau ntsuas cov khoom siv lead ua rau hauv lub raum tubular epithelial hlwb, MDCK hlwb (ntawm qhov ntom ntawm 2 × 105 hlwb / qhov zoo) tau cog thiab cog rau hauv txhua qhov dej ntawm 6-zoo phaj (Corning Inc.) rau 48 teev kom tau txais confluent monolayer. Lub cell monolayer tau incubated nrog FITC-labeled COM crystals (1000 µg crystal/ml nruab nrab) nyob rau hauv ib tug humidified incubator nrog 5 feem pua CO2 ntawm 37 ◦C rau 1 h. Tom qab ntawd, cov hlwb raug ntxuav nrog PBS thiab incubated nrog trypsin-EDTA tov kom tshem tawm cov uas tsis yog-internalized (ob adherent thiab tsis-adherent) crystals. Qhov feem pua ntawm cov hlwb nrog cov khoom siv sab hauv tau raug suav los ntawm tag nrho ntawm 10,000 qhov xwm txheej tau txais los ntawm kev siv lub cytometer ntws (BD Accuri C6) (BD Biosciences; San Jose, CA). Cov hlwb incubated nrog dawb (unlabelled) COM crystals tau siv rau presetting lub suab nrov thiab pib rau qhov zoo fluorescence teeb liab. Cov hlwb uas muaj cov teeb liab fluorescence zoo ces raug suav thiab siv rau kev suav feem pua.
2.8. COM crystal ntxeem tau los ntawm ECM kev soj ntsuam
COM crystals tau npaj raws li tau piav qhia saum toj no rau kev sib sau ua ke thiab siv lead ua-cell adhesion assays. Crystal invasion assay tau ua raws li cov txheej txheem tsim ua ntej [38,39]. Luv luv, tag nrho ntawm 20 µg COM crystals coated nrog crystallization buffer (kev tswj tsis pub dawb), DMSO (tsis zoo tswj), albumin (Sigma-Aldrich) (zoo tswj) los yogdiosmin(2.5, 5, 1{{10}}, 20, 40, 80 lossis 160 nM) tau ntxiv rau hauv 200 ul MEM. Tom qab ntawd, 200 ul ntawm 0.3 pM Lys-plasminogen (Fitzgerald Industries thoob ntiaj teb; Acton, MA) hauv PBS tau tov thiab incubated nrog crystal-protein complex ntawm 37 ◦C rau 1 h. Lub unbound plasminogen tau muab pov tseg los ntawm centrifugation ntawm 2000 g rau 5 min thiab cov pellet tau ntxuav nrog PBS ib zaug. Tom qab ntawd, 100 ul ntawm 0.15 pM urokinase plasminogen activator (uPA) (Fitzgerald Industries International) hauv PBS tau sib xyaw nrog crystal-protein-plasminogen / plasmin complex. Tom qab ntawd qhov sib tov tau ntxiv rau saum cov matrix gel hauv ECM migration chamber thiab incubated ntawm 37 ◦C. Tom qab 24-h incubation, cov tshuaj tseem nyob rau sab sauv ntawm lub migration chamber raug tshem tawm los ntawm kev siv cov ntaub qhwv los yog ntaub so ntswg. Lub invaded COM crystals nyob rau hauv lub matrix gel yog tom qab ntawd siv lub teeb tsom iav nrog qhov sib txawv cuam tshuam qhov sib txawv (DIC) hom (Nikon H600L). Cov siv lead ua ntxeem tau qhov deb tau ntsuas thiab ntsuas qhov nruab nrab ntawm tsawg kawg 15 qhov chaw qis zog (LPFs) hauv tib chav siv NIS Element D software version 4.11 (Nikon).
2.9. Kev txheeb cais
Tag nrho cov kev sim saum toj no tau ua tiav hauv triplicate (peb qhov kev sim ywj pheej) thiab cov ntaub ntawv ntau tau tshaj tawm raws li txhais tau tias ± SEM. Ntau qhov kev sib piv tau ua tiav siv ib txoj kev tsom xam ntawm qhov sib txawv (ANOVA) nrog Tukey's post-hoc test. Pearson correlation test tau ua los txiav txim siab txog kev sib raug zoo ntawm qhov sib txawv. Txhua qhov kev txheeb xyuas txheeb cais tau ua tiav los ntawm kev siv SPSS software (version 18) (IBM SPSS; Armonk, NY). P tus nqi tsawg dua 0.05 tau suav tias yog qhov tseem ceeb.
3. Cov txiaj ntsig
3.1. Cov nyhuv ntawm diosmin ntawm COM crystallization
Crystallization yog ib qho tseem ceeb ntawm cov kauj ruam thaum ntxovraumpob zeb tsim txheej txheem ntawm txhua hom. Tom qab 1-h crystallization nrog lossis tsis muajdiosminntawm ntau yam concentrations (2.5, 5, 10, 20, 40, 80 lossis 160 nM), COM siv lead ua loj thiab tus lej raug ntsuas thiab siv lead ua huab hwm coj tau suav. Crystallization tsis thiab diluent DMSO tau ua haujlwm raws li kev tswj hwm tsis zoo, raws li. Cov ntaub ntawv qhia tau hais tias tag nrho cov koob tshuaj (2.5–160 nM) ntawm disomin txo qis siv lead ua loj tab sis muaj cov siv lead ua ntau zuj zus nyob rau hauv cov koob tshuaj raws li qhov sib piv nrog cov kev tswj tsis zoo thiab tsis zoo (Fig. 1A–1C). Nyob rau hauv concordance nrog cov siv lead ua naj npawb, siv lead ua loj koob tshuaj-dependently nce (Fig. 1D). Zuag qhia tag nrho, cov ntaub ntawv no qhia tias diosmin txhawb COM crystallization (neocrystals).
3.2. Cov nyhuv ntawm diosmin ntawm COM siv lead ua kev loj hlob
Cov nyhuv modulatory ntawmdiosminntawm COM siv lead ua kev loj hlob tau soj ntsuam los ntawm kev ntsuas kev hloov pauv hauv cov siv lead ua loj (Δ Crystal loj) tom qab 60-min ntxiv incubation tom qab ua tiav cov theem pib crystallization, thaum neocrystals tsis tuaj. Cov txiaj ntsig tau pom tias tag nrho cov koob tshuaj (2.5–160nM) ntawm disomin txo qis Δ Crystal loj nyob rau hauv cov koob tshuaj raws li piv nrog rau kev tswj tsis pub dawb thiab tsis zoo (Daim duab 2). Cov kev tshawb pom no qhia tias diosmin inhibits COM crystal kev loj hlob.
3.3. Cov nyhuv ntawm diosmin ntawm COM crystal aggregation
Ntxiv nrog rau kev loj hlob siv lead ua, kev sib sau ua ke ntawm ib tus neeg siv lead ua uas ua kom nruj ua ke yog lwm kauj ruam tseem ceeb thaum lub sijhawmraumpob zeb pathogenesis. Qhov siab ntawm crystal aggregation thaum kawg tuaj yeem ua rau pob zeb loj thiab cuam tshuam ntawm lub raum me me tubular lumen. Sib piv nrog cov kev tswj tsis zoo thiab tsis zoo,diosminntawm 10–160nM koob tshuaj-raws li nce tus naj npawb ntawm COM crystal aggregates (Fig. 3). Cov txiaj ntsig no qhia tau tias diosmin txhawb COM crystal aggregation.
Cistanche rau lub raum ua haujlwm
3.4. Cov nyhuv ntawm diosmin ntawm COM crystal-cell adhesion
Kev khaws cia ntawm cov crystals causative yog ib qho ntawm cov kauj ruam tseem ceeb rauraumpob zeb tsim thiab tuaj yeem raug ntxias los ntawm crystal-cell adhesion uas tiv thaiv cov crystals los ntawm kev tshem tawm ua ke nrog cov zis tawm. Yog li peb ntsuas qhov kev ua haujlwm modulatory ntawmdiosminntawm COM crystal-cell adhesion. Cov ntaub ntawv qhia tau hais tias diosmin ntawm 5–160nM txo cov nplaum muaj peev xwm ntawm COM crystals ntawm lub raum tubular hlwb nyob rau hauv ib koob tshuaj raws li piv nrog rau cov dawb paug thiab tsis zoo tswj (Fig. 4). Cov ntaub ntawv no qhia tias diosmin inhibits COM crystal-cell adhesion.
3.5. Cov nyhuv ntawm diosmin ntawm COM crystal internalization rau hauv lub raum tubular hlwb
Tom qab adhesion, qee cov COM crystals tuaj yeem nkag mus rau hauv lub raum tubular hlwb thiab ua rau ob peb lub xovtooj tom ntej [37,40]. Crystal internalization tau soj ntsuam los ntawm kev siv FITC-labeled COM crystals thiab ntsuas los ntawm kev ntws cytometry. Plain COM crystals (tsis muaj labelling) tau siv los rho tawm cov suab nrov thiab xyuas kom meej tias cov fluorescence siv los ntawm FITC teeb liab. Muab piv nrog cov kev tswj tsis pub dawb thiab tsis zoo, feem pua ntawm cov hlwb nrog cov khoom siv hauv lub cev tau txo qis los ntawmdiosminntawm 10–160nM nyob rau hauv ib koob tshuaj raws li (Fig. 5). Cov kev tshawb pom qhia tias disomin inhibits COM crystal internalization rau hauv lub raum tubular hlwb.
3.6. Cov nyhuv ntawm diosmin ntawm COM crystal ntxeem tau los ntawm ECM
Crystal ntxeem tau los ntawm ECM yog cov txheej txheem pathogenic / kev puas tsuaj thaum lub sijhawmraumpob zeb pathogenesis uas tuaj yeem ua rau ntau yam inflammatory teb thiab cascades, yog li ua rau cov kab mob phem zuj zus. Peb tau tshuaj xyuas qhov tshwm sim no los ntawm kev siv cov txheej txheem tsim raws li kev ua haujlwm ntawm plasminogen-plasmin ntawm crystal-protein complex [38,39]. Cov txiaj ntsig tau pom tias tag nrho cov koob tshuaj ntawm disomin (2.5–160nM) tau txhim kho COM siv lead ua ntxeem tau los ntawm ECM raws li kev siv tshuaj (Daim duab 6). Txaus siab heev,diosminntawm 160nM tuaj yeem txhawb nqa COM siv lead ua ntxeem tau piv rau albumin, uas yog lub npe hu ua tus txhawb zog rau COM crystal ntxeem tau [41] (Fig. 6). Cov txiaj ntsig no qhia tias diosmin txhawb nqa COM crystal ntxeem tau los ntawm ECM.
Fig. 1. Cov nyhuv ntawmdiosminntawm COM crystallization. Crystallization assay tau ua nrog qhov sib npaug ntawm qhov ntim (4ul) ntawm crystallization buffer (kev tswj tsis pub dawb), DMSO (kev tswj tsis zoo) lossis diosmin (2.5–160nM). (A): Crystal morphology nyob rau hauv txhua yam mob tom qab 1-h crystallization. Thawj magnification yog 400 × rau tag nrho cov vaj huam sib luag. (B): Crystal loj. (C): Crystal tooj. (D): Crystal mass (saib Formula 1 hauv "Cov Khoom Siv thiab Cov Txheej Txheem") tau txheeb xyuas los ntawm tsawg kawg 100 crystals hauv 15 HPFs. Txhua bar sawv cev txhais tau tias ± SEM ntawm cov ntaub ntawv muab los ntawm 3 kev sim ywj pheej. *=p< 0.05="" vs.="" blank="" control;="" #="">< 0.05="" vs.="" dmso.="">
Fig. 2. Cov nyhuv ntawmdiosminntawm COM crystal kev loj hlob. Crystal kev loj hlob assay tau ua tom qab crystallization tiav (kom tiv thaiv neocrystallization) nrog qhov sib npaug ntawm qhov ntim (4ul) ntawm crystallization buffer (kev tswj tsis pub dawb), DMSO (kev tswj tsis zoo) lossis diosmin (2.5–160nM). (A): Crystal morphology nyob rau hauv txhua yam ntawm T0 thiab T60. Thawj qhov magnification yog 400 × rau tag nrho cov vaj huam sib luag. (B)-(J): Histograms ntawm crystal qhov ntau thiab tsawg ntsuas los ntawm ib tug neeg muaju ntawm T0 thiab T60 nyob rau hauv txhua pab pawg neeg. (K): Δ Crystal loj (saib Formula 2 hauv "Cov Khoom Siv thiab Cov Txheej Txheem") tau txheeb xyuas los ntawm tsawg kawg 100 crystals hauv 15 HPFs. Txhua bar sawv cev txhais tau tias ± SEM ntawm cov ntaub ntawv muab los ntawm 3 kev sim ywj pheej. *= p < 0.05="" vs.="" blank="" tswj;="" #="p">< 0.05="" vs.="">
Fig. 3. Cov nyhuv ntawmdiosminntawm COM crystal aggregation. Crystal aggregation assay tau ua nrog qhov sib npaug ntawm qhov ntim (4ul) ntawm crystallization buffer (kev tswj tsis pub dawb), DMSO (kev tswj tsis zoo) lossis diosmin (2.5–160nM). (A): Micrographs ntawm aggregated COM crystals (labeled nrog dotted voj voog). Thawj magnification yog 400 × rau tag nrho cov vaj huam sib luag. (B): Tus naj npawb ntawm cov crystal aggregates raug suav los ntawm tsawg kawg yog 15 random HPFs hauv txhua qhov dej. Txhua bar sawv cev txhais tau tias ± SEM ntawm cov ntaub ntawv muab los ntawm 3 kev sim ywj pheej. *=p< 0.05="" vs.="" blank="" control;="" #="p" <="" 0.05="" vs.="" dmso.="">
Fig. 4. Cov nyhuv ntawmdiosminntawm COM crystal-cell adhesion. Crystal-cell adhesion assay tau ua nrog qhov sib npaug ntawm qhov ntim (4ul) ntawm crystallization buffer (kev tswj tsis pub dawb), DMSO (kev tswj tsis zoo) lossis diosmin (2.5–160nM). (A): Micrographs ntawm cov pob zeb uas tseem tshuav uas nruj nreem rau ntawm lub cell monolayer tom qab tshem tawm cov pob zeb tsis muaj zog los ntawm kev ntxuav tes nrog PBS. Thawj magnification yog 200 × rau tag nrho cov vaj huam sib luag. (B): Tus naj npawb ntawm cov khoom siv lead ua tau raug suav los ntawm tsawg kawg 15 qhov chaw nyob hauv txhua qhov dej. Txhua bar sawv cev txhais tau tias ± SEM ntawm cov ntaub ntawv muab los ntawm 3 kev sim ywj pheej. *= p<0.05 vs.="" blank="" control;="" #="p"><0.05 vs.="">
Fig. 5. Cov nyhuv ntawmdiosminntawm COM crystal internalization rau hauv lub raum tubular hlwb. Crystal internalization assay tau ua tiav siv FITC-labeled COM crystals (FITC-COM), whereas cov dawb (tsis-labeled) COM crystals tau siv rau keeb kwm yav dhau / suab rho tawm. Qhov kev ntsuam xyuas tau ua nrog qhov sib npaug ntawm qhov ntim (4ul) ntawm crystallization buffer (kev tswj tsis pub dawb), DMSO (kev tswj tsis zoo) lossis diosmin (2.5–160nM). (A): Flow cytometric dot-plot tsom xam ntawm qhov loj (y-axis) thiab FITC-fluorescence siv (x-axis) ntawm cov hlwb tom qab tshem tawm cov crystals uas tsis yog internalized nrog 0.1 feem pua trypsin / 2.5mM EDTA. (B): Feem pua ntawm cov hlwb nrog FITC-labeled COM crystals. Txhua bar sawv cev txhais tau tias ± SEM ntawm cov ntaub ntawv muab los ntawm 3 kev sim ywj pheej. *= p<0.05 vs.="" fitc-com="" +="" blank="" control;="" #="">< 0.05="" vs.="" fitc-com="" +="" dmso.="">
Fig. 6. Cov nyhuv ntawmdiosminntawm COM crystal ntxeem tau los ntawm ECM. Crystal invasion los ntawm ECM kev soj ntsuam tau ua tiav siv COM crystals coated nrog crystallization tsis (kev tswj tsis pub dawb), DMSO (tsis zoo tswj), albumin (zoo tswj) lossis diosmin (2.5–160nM). (A): Micrographs ntawm COM crystals invaded los yog tsiv teb tsaws los ntawm ECM migration chamber. Thawj magnification yog 100 × rau tag nrho cov vaj huam sib luag. (B): Crystal invasion nrug tau ntsuas los ntawm tsawg kawg 15 random LPFs nyob rau hauv txhua chav. Txhua bar sawv cev txhais tau tias ± SEM ntawm cov ntaub ntawv muab los ntawm 3 kev sim ywj pheej. *= p<0.05 vs.="" blank="" control;="" #="p" <="" 0.05="" vs.="">
4. Kev sib tham
Kab mob raum pob zeb yog tshwm sim los ntawm kev tsim cov calculi nyob rau hauv lubraumthiab pelvocalyceal system. Cov txheej txheem ntawm lub raum pob zeb tsim muaj xws li COM crystallization, kev loj hlob, kev sib sau ua ke, khaws cia los ntawm crystal-cell adhesion, thiab ntxeem tau los ntawm lub raum interstitium nplua nuj nrog ECM [21,22]. Muaj ntau qhov kev sim los tiv thaiv tus kab mob no siv ntau yam tshuaj thiab / lossis cov khoom noj khoom haus. Ntau cov ntaub ntawv pov thawj tsis ntev los no tau tshaj tawm tias flavonoids, flavone glycosides thiab lwm cov tshuaj uas tau muab rho tawm los ntawm citrus txiv hmab txiv ntoo tuaj yeem muaj kev tiv thaiv kab mob raum pob zeb thiab lwm yam kab mob [23–25]. Ntawm cov no,diosmin, lub ntuj flavone glycoside thiab hesperidin derivative pom nyob rau hauv cov txiv hmab txiv ntoo citrus [1,2], tau raug pom zoo los ua lub luag haujlwm tiv thaiv kev raug mob thiab kev puas tsuaj ntawmraumcev [13,42]. Ntxiv mus, nws lub luag haujlwm tiv thaiv urolithiasis tau tshaj tawm hauv ob peb txoj kev tshawb fawb yav dhau los siv cov qauv tsiaj [26,27]. Txawm li cas los xij, cov txiaj ntsig tau zoo thiab cov txheej txheem hauv qab ntawm diosmin hauv kev tiv thaiv lub raum pob zeb tseem hawb pob. Yog li peb soj ntsuam cov kev ua haujlwm modulatory ntawm diosmin ntawm COM crystals. Cov kev tshuaj ntsuam xyuas tau ua tiav nyob rau ntau theem ntawm COMraumpob zeb tsim, suav nrog crystallization, siv lead ua kev loj hlob, sib sau ua ke, siv lead ua-cell adhesion, internalization rau hauv lub raum tubular hlwb thiab ntxeem tau los ntawm ECM.
Tom qab noj qhov ncaujdiosmin, nws qib plasma hauv tib neeg yog nyob ntawm {{0}}.5 txog 200ng/ml (los yog 0.8–300nM) [43,44]. Qhov siab tshaj plaws plasma concentration (Cmax) yog kwv yees li 50ng / ml (los yog 85nM) [43,44]. Yog li, qhov ntau npaum li cas ntawm diosmin siv hauv peb txoj kev tshawb fawb tam sim no (2.5-160nM) yog nyob rau hauv cov tshuaj muaj feem cuam tshuam rau nws cov tshuaj pharmacokinetics. Vim tias DMSO tau siv los ua cov diluent kom yaj tag nrho diosmin ib qhov kev pom zoo, DMSO tau siv los ua qhov kev tswj tsis zoo hauv qhov kev tshawb fawb no ntxiv rau kev tswj tsis pub dawb kom ntseeg tau tias tsis muaj kev cuam tshuam los ntawm diluent nws tus kheej uas yuav cuam tshuam nrog cov ntaub ntawv txhais lus. Hauv txhua qhov kev ntsuam xyuas, cov ntaub ntawv tau pom tias tsis muaj qhov cuam tshuam loj heev los ntawm DMSO tau pom thiab tag nrho cov ntaub ntawv ntau tau los ntawm kev tswj tsis zoo tau muab piv rau cov kev tswj tsis pub muaj.
COM crystallization assay qhia tias tag nrho cov concentrations ntawmdiosmin(2.5, 5, 10, 20, 40, 80 thiab 160nM) tuaj yeem txo COM siv lead ua loj tab sis, ntawm qhov tod tes, nce tus naj npawb ntawm cov muaju. Raws li cov khoom siv lead ua yog cov khoom kawg ntawm ob qho tib si siv lead ua loj thiab tus naj npawb thiab muaj feem cuam tshuam ntau dua los cuam tshuam cov qib ntawm COM crystallization, peb mam li ntsuam xyuas seb diosmin cuam tshuam qhov ntsuas qhov ntsuas no. Cov ntaub ntawv qhia tau hais tias tag nrho cov concentrations ntawm diosmin muaj kev txhawb nqa ntawm COM siv lead ua loj, raws li cov ntaub ntawv ntawm crystal tooj. Tom qab crystallization, COM crystals tuaj yeem loj hlob mus rau qhov loj dua rau cov kauj ruam tom ntejraumpob zeb tsim.
Nyob rau hauv sib piv rau crystallization, diosmin ntawm tag nrho cov concentrations nthuav tawm cov nyhuv inhibitory ntawm COM siv lead ua kev loj hlob nyob rau hauv cov koob tshuaj raws li. Qee qhov kev tshawb fawb yav dhau los tau tshaj tawm tias kev solubility ntawm CaOx muaju tuaj yeem nce ntxiv los ntawm cov khoom siv ntawm hydroxyanthraquinones nrog glycosylation [45]. Tsis tas li ntawd, muaj cov suab thaj nyob rau hauv xws li bioactive tebchaw tuaj yeem khi nrog dawb calcium ion tej zaum vim yog cov pawg hydroxyl hauv cov qauv molecular, yog li inhibiting tsim ntawm CaOx crystals [45,46]. Diosmin kuj tseem tuaj yeem cuam tshuam rau kev hloov pauv ntawm cov calcium uas solubilized thiab oxalate ions mus rau hauv COM crystalline hais nyob rau hauv lub raum tubular kua nyob rau hauv tus txheej txheem ntawm COM crystallization raws li cov txheej txheem no.
Txawm li cas los xij, COM crystal aggregation tau nce los ntawm 10-160nMdiosmin, implicating lub peev xwm ntawm diosmin los ua ib tug linker los yog nplaum molecule rau recruiting ib tug neeg muaju rau khi ua ke los tsim cov siv lead ua aggregates. Qhov kev khi no tseem ua rau muaj kev nce qib ntawm cov nplaum txuas ntawm cov neeg siv lead ua thiab tuaj yeem ua rau cov qauv loj ntawm COM muaju, uas yooj yim ua rau cov pob zeb nidus tso rau hauvraum[32].
COM siv lead ua tuav los ntawm qhov adhesion ntawm crystals ntawm lub raum tubular cell nto tau tsim los ua lwm yam tseem ceeb rau lub raum pob zeb tsim [47]. Peb cov ntaub ntawv tau qhia tias diosmin tuaj yeem txo cov nplaum nplaum ntawm COM crystals los khi nrog apical nto ntawm MDCK raum tubular hlwb. Qee qhov kev tshawb fawb yav dhau los tau pom tias glycosaminoglycans (polysaccharide compounds) coated ntawm CaOx crystals los yog qhia ntawm lub raum tubular hlwb tuaj yeem cuam tshuam nrog cov nplaum muaj peev xwm ntawm CaOx crystals mus rau lub raum tubular hlwb [48]. Tsis tas li ntawd, qhov kev qhia ntawm crystal receptors ntawm lub raum tubular hlwb yog ib qho ntawm cov txheej txheem tseem ceeb uas txiav txim siab siv lead ua-cell adhesion [21]. Thaum CaOx crystals induces raug mob ntawm apical daim nyias nyias, tshwj xeeb tshaj yog microvilli, qee cov polyphenols xws li epigallocatechin gallate (EGCG) tuaj yeem tiv thaiv cov crystal-cell adhesion thiab cellular raug mob [49]. Ntxiv mus, cov tshuaj flavonoid nyob rau hauv ntau yam nroj tsuag rho tawm tau tshaj tawm yav dhau los kom nce pH theem ntawm cov zis, ua rau inhibition ntawm crystal-cell adhesion [50,51]. Tej zaum,diosminkuj tseem tuaj yeem txo qhov kev raug mob ntawm tes los ntawm COM, yog li txo cov crystal-cell adhesion.
COM siv lead ua internalization tau pom tias tshwm sim los ntawm endocytosis ntawm actin cytoskeleton-mediated macropinocytosis txoj kev [40]. Flavonoids xws li quercetin tau tshaj tawm tias cuam tshuam rau actin cytoskeleton xav tau rau macropinocytosis [52,53]. Peb qhov kev tshawb pom tau pom tias diosmin txo qis lub peev xwm ntawm MDCK hlwb rau hauv COM crystals. Cov nyhuv inhibitory ntawm diosmin tej zaum yuav cuam tshuam nrog kev sib dhos lossis lub koom haum ntawm actin cytoskeleton hauv cov hlwb uas cuam tshuam ncaj qha rau txoj kev macropinocytosis [54,55].
Tom qab internalization, COM crystals tau degraded los ntawm endolysosomes uas ua rau kom cov calcium dawb thiab oxalate ions nyob rau hauv lub raum interstitium [37]. Xws li kev nce hauv calcium thiab oxalate ions tuaj yeem ua rau COM neocrystallization hauv lub raum interstitium [56,57]. Tsis tas li ntawd, lub xub ntiag ntawm interstitial COM crystals tej zaum yuav yog los ntawm qhov tsis xws luag ntawm kev sib tshuam nruj thiab paracellular adhesion barriers, ua rau muaj zog paracellular permeability thiab siv lead ua translocation [58-60]. Cov crystals no tuaj yeem cuam tshuam rau lub raum interstitium los ntawm ECM thiab tom qab ntawd ua rau ntau qhov inflammatory teb thiab cov ntaub so ntswg puas [58–60]. Hauv txoj kev tshawb no, peb tau pom tias kev cuam tshuam ntawm COM crystals los ntawm ECM tsiv teb tsaws chamber raug ntxias los ntawm tag nrho cov concentration ntawm diosmin. Diosmin tej zaum yuav khi rau COM siv lead ua qhov chaw thiab cuam tshuam nrog plasminogen-plasmin system uas tsav cov siv lead ua migration hauv ECM migration chamber [38,39].
Cistanche rau lub raum
Vimdiosmininduced ob qho tib si inhibiting thiab txhawb cov teebmeem ntawm ntau yam txheej txheem ntawm COM pob zeb tsim, peb thiaj li txiav txim siab lawv txoj kev sib raug zoo siv Pearson correlation test. Kev txheeb xyuas kev sib raug zoo tau qhia tias tus lej siv lead ua sib cuam tshuam nrog ob qho tib si siv lead ua loj thiab siv lead ua loj hlob (Δ Crystal loj) (Daim duab 7A thiab B). Crystal loj sib cuam tshuam nrog kev loj hlob siv lead ua (Fig. 7C) tab sis inversely correlated nrog crystal mass (Fig. 7D). Thaum kawg, cov khoom siv lead ua muaj kev sib raug zoo nrog cov lej siv lead ua thiab cov khoom siv lead ua ke (Fig. 7E thiab F). Los ntawm kev txheeb xyuas kev sib raug zoo, cov ntaub ntawv qhia tias muaj pes tsawg tus siv lead ua thiab cov khoom siv lead ua muaj feem cuam tshuam rau kev cuam tshuam crystallization ntau dua li siv lead ua loj (Fig. 7A, D thiab E). Tsis tas li ntawd, qhov luaj li cas ntawm neocrystals thaum lub sij hawm crystallization muaj feem xyuam rau kev loj hlob ntawm cov preformed crystals (Fig. 7C), whereas degree ntawm crystallization (raws li reflected los ntawm crystal nab npawb thiab crystal mass) yog ze ze rau degree ntawm crystal aggregation (Fig. 7E thiab. F). Txawm li cas los xij, cov kev sib raug zoo no feem ntau yog tshwj xeeb rau cov teebmeem diosmin, qhov cuam tshuam los ntawm lwm cov modulators tuaj yeem lossis tsis muaj qhov sib xws. Piv txwv li, fibronectin txo cov khoom siv lead ua thiab inhibits crystal kev loj hlob, tab sis txhawb nqa crystal aggregation [31]. Tsis tas li ntawd, tsis muaj zog Escherichia coli nce siv lead ua loj thiab loj tab sis tsis muaj kev cuam tshuam rau cov lej siv lead ua [33]. Cov ntaub ntawv no qhia tau hais tias kev sib raug zoo ntawm ntau yam COM crystal assay tsis yog universal rau tag nrho cov modulators.
Thaum kawg, peb tau sau cov ntsiab lus tau txais los ntawm ntau qhov kev ntsuam xyuas thiab sib xyaw ua ke nrog cov txheej txheem pathogenic ntawmraumpob zeb kab mob (saib schematics hauv daim duab 8). Los ntawm kev txheeb xyuas kev sib raug zoo, diosmin txhawb nqa crystallization (Fig. 8A). Nruab nrab ntawm crystallization thiab crystal kev loj hlob,diosminua kom muaj kev sib npaug zoo los ntawm kev txhawb nqa crystallization tab sis, ntawm qhov tod tes, inhibiting crystal kev loj hlob (Fig. 8B). Hais txog tag nrho cov teebmeem ntawm COM crystals ib leeg (tsis xav txog cov hlwb thiab ECM uas tseem cuam tshuam), kev txhawb nqa kev ua haujlwm ntawm diosmin yog qhov tseem ceeb tshaj li nws cov haujlwm inhibiting ntawm COM crystals vim nws txhawb nqa crystallization thiab crystal aggregation, tab sis inhibits tsuas yog siv lead ua kev loj hlob ( Daim duab 8C). Txawm hais tias crystallization nce, qhov loj me ntawm crystallization txo thiab cuam tshuam nrog kev loj hlob inhibition (Daim duab 7C). Cov ntaub ntawv no zoo ib yam nrog cov txiaj ntsig tau tshaj tawm los ntawm Kavanagh li al. [61–63], ua kom pom tias qhov nce ntawm crystallization ua rau poob ntawm supersaturation ntawm calcium thiab oxalate ions hauv cov tshuaj, yog li txo qhov kev loj hlob. Txawm li cas los xij, kev loj hlob siv lead ua tsis yog tib qho kev txiav txim siabraumpob zeb pathogenesis [64], tshwj xeeb tshaj yog thaum cov siv lead ua loj overwhelms thiab cuam tshuam nrog kev nce ntawm cov siv lead ua aggregation (Fig. 7F) uas tuaj yeem ua rau muaj kev pheej hmoo ntawm cov khoom siv lead ua kom tau daig hauv me me tubular ntu (hauv qhov kev xav ntawm intratubular), yog li nce ntxiv. lub caij nyoog ntawm pob zeb tsim. Tsis tas li ntawd, ib txoj kev tshawb fawb yav dhau los tau qhia cov ntaub ntawv raws li peb qhov kev tshawb pom, qhia tias qhov nce ntawm cov siv lead ua muaj feem cuam tshuam nrog kev nce ntawm cov khoom siv lead ua thiab cov pob zeb loj [63].
Daim duab 7. Kev txheeb xyuas kev sib txheeb. (A)–(F): Kev sib raug zoo ntawm COM siv lead ua tooj, siv lead ua loj, Δ Crystal loj, siv lead ua loj, thiab ib tug xov tooj ntawm cov siv lead ua aggregates raug soj ntsuam los ntawm Pearson correlation test.
Daim duab 8. Schematics summarizing lub modulatory teebmeem ntawmdiosminntawm COMraumcov txheej txheem tsim pob zeb. (A): Cov teebmeem ntawm diosmin ntawm COM crystallization. (B): Cov teebmeem ntawm diosmin ntawm COM crystallization thiab siv lead ua loj hlob. (C): Cov teebmeem ntawm diosmin ntawm COM crystallization, kev loj hlob thiab kev sib sau ua ke. (D): Cov teebmeem ntawm diosmin rau tag nrho COM raum pob zeb txheej txheem.
Thaum muaj kev cuam tshuam nrog lub raum tubular hlwb thiab ECM raug txiav txim siab, lub ntiaj teb daim duab ntawm ob qhov teebmeem ntawmdiosminntawm COM lub raum pob zeb tsim ua kom pom tseeb dua (Fig. 8D). Diosmin inhibits crystal-cell adhesion, yog li txo cov crystal internalization rau hauv lub hlwb. Qhov no tuaj yeem piav qhia tias qhov me me ntawm COM crystals muaj peev xwm ua kom tsis muaj zog los khi lub raum tubular hlwb piv nrog cov loj dua li tau tshaj tawm hauv peb txoj kev tshawb fawb tsis ntev los no [34] thiab kev kawm dhau los los ntawm lwm pab pawg [65]. Qhov tsis muaj peev xwm ua kom muaj peev xwm ntawm cov COM me me yog vim lawv cov nplaum tsis tshua muaj zog rau ntawm lub xov tooj ntawm tes thiab tsawg tus lej ntawm COM siv lead ua receptors khi rau lawv raws li kev txiav txim siab los ntawm atomic force microscopy thiab proteome analysis, feem [34]. Diosmin kuj inhibits crystal internalization. Nco ntsoov tias cov txheej txheem internalization yog ob-edged ntaj uas yuav tsum tau ceev faj txhais. Internalization los yog endocytosis nyob rau hauv ib qho ntawm cov kev tiv thaiv mechanisms uas hlwb siv rau kev tshem tawm cov muaju los ntawm endolysosomes. Txawm li cas los xij, degradation ntawm cov pob zeb tsis zoo tsim cov calcium dawb thiab oxalate ions uas tuaj yeem txav los ntawm intracellular compartment mus rau lub raum interstitium, uas lawv tuaj yeem tsim cov neocrystals [56,57]. Ntawm qhov tod tes, diosmin txhawb nqa crystal ntxeem tau los ntawm ECM, uas yog ib qho ntawm cov txheej txheem tseem ceeb rau lub raum pob zeb pathogenesis (tshwj xeeb hauv Randall's plaque qauv) [58–60]. Zuag qhia tag nrho, Fig. 8D qhia txog tag nrho cov kev hloov pauv dual ntawm diosmin ntawm COMraumcov txheej txheem tsim pob zeb. Nws yuav tsum raug sau tseg tias qhov sib npaug ntawm cov kev txhawb nqa thiab inhibiting cov teebmeem no tsis tuaj yeem suav tau meej (tsis zoo li kev ua lej). Ntxiv mus, muaj ob peb lwm yam endogenous thiab exogenous yam uas tuaj yeem cuam tshuam cov txheej txheem tsim pob zeb. Thaum kawg, diosmin kuj nthuav tawm ntau qhov cuam tshuam ncaj qha rau lub raum pob zeb tsim, suav nrog nws cov tshuaj tiv thaiv antioxidant thiab tiv thaiv kab mob [5,6] uas yuav tsum tau coj mus rau hauv tus account. Yog li ntawd, qhov kawg tshwm sim ntawm no dual modulatory teebmeem ntawmdiosminntawm COMraumkev tsim pob zeb xav tau ntxiv hauv vivo kev tshawb nrhiav thiab kev tshawb fawb loj-cohort yav tom ntej.
5. Cov lus xaus
Hauv cov ntsiab lus, peb tshaj tawm ntawm no nyob rau hauv ob qhov teebmeem ntawm diosmin ntawm COM siv lead ua kev hloov pauv nyob rau hauv ib koob tshuaj. Thaum nws inhibits COM siv lead ua kev loj hlob, crystal-cell adhesion, thiab internalization rau hauv lub raum tubular hlwb,diosmintxhawb COM crystallization, aggregation, thiab ntxeem tau los ntawm ECM. Yog li ntawd, nws lub luag haujlwm los tiv thaiv urolithiasis yog qhov tsis ntseeg thiab yuav tsum tau ceev faj txog nws txoj kev siv rau cov kab mob raum pob zeb.
CREDiT daim ntawv qhia txog kev sau ntawv
Supaporn Khamchun: Conceptualization, Methodology, Software, Validation, Formal analysis, Investigation, Data curation, Writing – original draft, Visualization, Sunisa Yoodee: Conceptualization, Methodology, Software, Validation, Formal analysis, Investigation, Data curation, Sau ntawv – thawj daim ntawv, Visualization, Mus ntsib Thongboonkerd: Conceptualization, Methodology, Software, Validation, Resources, Writing – review & editing, Supervision, Project Administration, Funding acquisition.
Kev tsis sib haum xeeb ntawm kev txaus siab nqe lus
Cov kws sau ntawv tshaj tawm tsis muaj teeb meem ntawm kev txaus siab.
Kev lees paub
Peb ua tsaug rau kev pab cuam ntawm Kittiya Suwannakud. Txoj haujlwm no tau txais kev txhawb nqa los ntawm Lub Chaw Haujlwm Saib Xyuas Kev Tshawb Fawb Txog Kev Tshawb Fawb Hauv Tebchaws thiab Kev Tsim Kho Tshiab (NXPO) los ntawm PMU-B thiab Thaib Kev Tshawb Fawb Nyiaj Txiag (IRN60W0004). VT kuj tau txais kev txhawb nqa los ntawm "Chalermphrakiat" Grant, Kws Qhia Ntawv ntawm Tshuaj Kho Mob Siriraj Tsev Kho Mob.
Cov ntaub ntawv
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