Calcium kho li cas rau kev puas tsuaj rau lub raum luoride?
Mar 16, 2022
Hu rau: Audrey Hu Whatsapp / hp: 0086 13880143964 Email:audrey.hu@wecistanche.com
Ntsiab lus: Calcium, Lub raum, Apoptosis
TSAB NTAWV
Kev noj tshuaj fluoride (F) ntau dhau mus ntev tuaj yeem ua rau osseous thiab tsis-osseous puas. Covraumyog lub ntsiab fluoride excretion kab mob ntawm lub cev. Txoj kev tshawb no tsom los tshawb xyuas seb puas noj zaub movcalciumsupplementation yuav alleviateraumkev puas tsuaj los ntawm fluorosis thiab mus soj ntsuam ntxiv cov teebmeem ntawmCalciumntawm lub mitigation mechanism ntawmraumcellapoptosistshwm sim los ntawm F. Peb soj ntsuam cov qauv histopathological,raumcov ntsuas kev ua haujlwm, thiab cov noob thiab cov protein qhia theem ntawm kev tuag receptor-mediatedapoptosistxoj hauv kev hauv Sprague Dawley (SD) nas kho nrog sodium fluoride (NaF) thiab/los yogcalciumcarbonate (CaCO3) rau 120 hnub. Cov txiaj ntsig tau pom tias 100 mg / L NaF inducedraumhistopathological raug mob thiabapoptosis, nce cov concentrations ntawm Creatinine (CRE), uric acid (UA), ntshav urea nitrogen (BUN), poov tshuaj (K), phosphorus (P), thiab F (p < {{0}}}.05)="" ,="" thiab="" txo="" qib="" ntawm="" cov="" piam="" thaj="" magnesium="" (mg)="" (p=""><0.05). ntxiv="" mus,="" naf="" tau="" nce="" mrna="" thiab="" cov="" protein="" qhia="" theem="" ntawm="" fas="" cell="" nto="" tuag="" receptor="" (fas),="" qog="" necrosis="" factor="" (tnf),="" tnf="" ntsig="">apoptosis-inducing ligand (TRAIL), Caspase 8, Caspase 3, thiab poly ADP-ribose polymerase (PARP) (p < 0.01),="" uas="" thaum="" kawg="" ua="" rau="" txoj="" kev="" tuag="" receptor.="">Calciumsupplementation reversed qhov txo qis ntawm CRE, BUN, UA, F, thiab P theem induced los ntawm F, alleviated histopathological puas thiabapoptosis, thiab txo cov noob thiab cov protein qhia theem ntawm kev tuag receptor txoj kab ntsig txog cov cim. Hauv kev xaus, 1 feem puaCalciumalleviates F-inducedraumapoptosislos ntawm FAS/FASL, TNFR/TNF, DR5/TRAIL signaling pathways.
Taw qhia
Fluorine yog dav nyob rau hauv av, dej, thiab zaub mov. Txawm li cas los xij, qee qhov xwm txheej ntuj (tej huab cua ntawm cov zaub mov thiab pob zeb, tsim ntawmcalciumthiab magnesium ntsev, infiltration ntawm dej hauv av, thiab lwm yam) thiab tib neeg kev ua ub no (kev lag luam dej khib nyiab, agrochemicals, khoom siv hauv tsev, thiab lwm yam) ua rau muaj kuab paug fluorine hauv ib puag ncig (Daiwile li al., 2019). Tam sim no, qhov loj tshaj plaws ntawm kev raug tshuaj fluoride (F) yog cov dej haus, thiab qhov pom zoo ntawm F hauv cov dej haus los ntawm World Health Organization yog tsawg dua 1.5 mg / L (Cov Lus Qhia rau Kev Haus Dej Haus, 2017). Cov pov thawj ntawm kev kis kab mob qhia tau hais tias siab dua qhov concentration no yuav ua rau muaj kev pheej hmoo ntawm kev kho hniav fluorosis, thiab qhov nce ntxiv yuav ua rau muaj kev pheej hmoo ntawm pob txha fluorosis (Cov Lus Qhia rau Kev Haus Dej- Dej Zoo, 2017; National Research Council, 2006). Lub sij hawm ntev ua rau muaj cov tshuaj fluoride ntau tuaj yeem ua rau lub cev nqus tau cov tshuaj fluorine los ntawm kev ua pa.
thiab digestive ib ntsuj av. Kev noj ntau dhau ntawm F tuaj yeem ua rau cov pob txha thiab cov pob txha tsis zoo ntawm lub cev. Cov ntaub so ntswg puas sab nraum cov pob txha feem ntau hu ua kev puas tsuaj uas tsis yog pob txha. Tam sim no, nws tau pom tias kev puas tsuaj ntawm cov ntaub so ntswg uas tsis yog pob txha tshwm sim los ntawm F cuam tshuam nrog kev zom zaub mov, daim siab,raum, hlwb, thiab lwm yam. (Jha et al., 2011; Perumal et al., 2013).
Covraum, lub ntsiab excretory kab mob ntawm lub cev, yog lub luag hauj lwm rau lub metabolism ntawm cov tshuaj lom thiab exogenous toxins nyob rau hauv lub cev. Cov kev tshawb fawb tau tshaj tawm tias kwv yees li 50-80 feem pua ntawm cov tshuaj fluoride noj los ntawm lub cev yog lim thiab rov qab los ntawm cov tshuaj fluoride.lub raum, thiab cov tshuaj fluorine ntxiv yog khaws cia rau hauv lwm cov ntaub so ntswg thiab lub cev (Chen li al., 2013; Dharmaratne, 2019). Ntau hom ntaub ntawv tau pom tias 50 thiab 100 mg / LF raug tuaj yeem ua rau lub raum kab noj hniav loj, atrophy ntawmraumtubules, irregular txheej ntawm papillary hlwb, nqaim ntawm lub lumen, ua rauapoptosisntawmraumcell thiab deterioration ntawm biochemical functions xws li creatinine (CRE), uric acid (UA),calcium(Ca), thiab phosphorus (P) (Song et al., 2014; HW Wang et al., 2020; Wei et al., 2018a).
Apoptosis, ib hom cell tuag tswj los ntawm cov noob, uas muab faib ua endogenousapoptosisthiab exogenousapoptosis. Cov ntawv ceeb toom tsis ntev los no tau qhia tiasapoptosistshwm sim los ntawm F siab feem ntau suav nrog mitochondria-mediated, endoplasmic reticulum yog kev ntxhov siab-kho, thiab txoj kev tuag receptor-mediated pathways (Wei li al., 2018a). Cov kev sim yav dhau los hauv peb pab pawg tau taw qhia tias NaF ua rau cov pob txha thiab lub siabapoptosislos ntawm txoj kev intracellular endoplasmic reticulum (ER) thiab mitochondrial txoj kev (Li et al., 2021; J. Wang li al., 2020; Wang et al., 2019). Tsis tas li ntawd, cov kev tshawb fawb kuj tau pom tias txoj kev mitochondrial tau koom nrog hauv NaF-inducedapoptosisntawm naslub raum(Wei et al., 2018b). Txawm li cas los xij, tsis muaj kev tshawb fawb txog kev tuag receptor-koom haum txoj hauv kev hauv F-inducedraumapoptosis. Txoj kev tuag receptor dominates txhua theem ntawm exogenousapoptosis, suav nrog Fas cell surface death receptor (FAS) pathway, qog necrosis factor (TNF) pathway, thiab TNF-relatedapoptosis-inducing ligand (TRAIL) txoj kev (Grunert li al., 2012; Lu li al., 2017; Wang thiab Su, 2018). FAS txoj hauv kev yog lub cim tseem ceeb ntawm qhov kev hloov pauv uas cuam tshuamapoptosis. FAS muaj cov yam ntxwv ntawm daim nyias nyias receptor thiab tsim ib lub trimer tom qab khi nrog FAS ligand (FAS-L). Tom qab ntawd qhov tshwj xeeb ntawm lub trimer khi nrog tus neeg tuag (DD) ntawm qhov sib txuas connexin molecule FAS-associated death domain protein (FADD) los tsim ib qho kev tuag-induced signal complex (DISC) (Wang thiab Su, 2018). Hauv cov txheej txheem kev hloov pauv roj ntsha hauv qab no, FADD tuaj yeem qhib Caspase 8 thiab Caspase tsev neeg tib lub sijhawm, thiab thaum kawg ua rauapoptosislos ntawm kev koom tes ntawm Caspase 3 thiab poly ADP-ribose polymerase (PARP) nyhuv (Kischkel li al., 2000; Meynier thiab Rieux-Laucat, 2019). Cov kev tshawb fawb tsis ntev los no tau pom tias FAS txoj kev sib khoapoptosisnyob rau hauv T cell thiab thaum lub sij hawmraumkev ua tsis tiav los ntawm cisplatin (Djiadeu li al., 2017; Linkermann li al., 2011). Tsis tas li ntawd, ib txoj kev tshawb fawb kuj tau tshaj tawm tias NaF inducesapoptosisnyob rau hauv nas los ntawm txoj kev FAS (Sun et al., 2017). TRAIL yog ib qho kev lees paub cytokine hauv TNF superfamily. Nws khi rau nws homologous agonist receptors, uas yog kev tuag receptors (DR4 thiab DR5). Muaj DD hauv nws cheeb tsam cellular. DD yog qhov tsim nyog rau kev nrhiav neeg ua haujlwm hloov pauv protein FADD. FADD nyob rau hauv lem induces tso tawm Caspases mus rau hauv lub cytoplasm, activated Caspase 3 cleaves PARP, thiab inhibits nws DNA kho muaj peev xwm, ua rau lubapoptosis(Bodmer et al., 2000; Micheau, 2018). Nws tau raug qhia tias txoj kev TRAIL/DR5 tau koom nrog hauv NaF-inducedapoptosishauv nas (Song et al., 2021). Osteoprotegerin (OPG) yog ib qho dawb soluble receptor uas tsis muaj qhov chaw hloov pauv. Nws tuaj yeem txwv txoj kev sib haum xeebapoptosislos ntawm inhibiting kev khi ntawm TRAIL thiab lwm yam kev tuag receptors (Duiker li al., 2006; Kiraz li al., 2016; Micheau, 2018). Kev khi ntawm TNF thiab TNF receptor (TNF-R1) tuaj yeem qhib kev nrhiav neeg ua haujlwm ntawm TNFR-sociated death domain (TRADD) protein los ntawm nws cov DD. Tom qab ntawd, TRADD cuam tshuam nrog FADD, ua rau kev nrhiav neeg ua haujlwm ntawm Pro-Caspase 8, uas yog cleaved rau hauv active Caspase 8 los ntawm proteolytic enzymes. Caspase 8 ces activates Caspase 3, uas yog lub luag hauj lwm rau cellapoptosis(Kiraz li al., 2016; Sedger thiab McDermott, 2014). Nws tau tshaj tawm tias NaF induces hepatocyteapoptosisnyob rau hauv nas los ntawm TNF-R1 teeb liab txoj kev (Lu li al., 2017)
Calciumplays ntau lub luag haujlwm hauv cov pob txha thiab cov hniav, thiab nws kuj tuaj yeem tswj cov hlab ntsha thiab cov khoom tso tawm (Cao li al., 2016), koom nrog hauv kev ua haujlwm.calciumhomeostasis (Carmeliet li al., 2003; Yang li al., 2016), hloov membrane permeability (Lappe li al., 2017), qhib lub zais ntawm ntau yam enzymes thiab cov tshuaj hormones (Kim li al., 2012), thiab lwm yam. Ntau qhov kev tshawb fawb yav dhau los tau pom tias F thiab Ca muaj qhov cuam tshuam tsis zoo hauv biology (Dure-Smith li al., 1996; Nobrega li al., 2019). Thaum F yog absorbed los ntawm lub cev, nws yuav nkag mus rau hauv cov ntshav los tsim insoluble CaF2 precipitates. Yog tias kev noj zaub mov ntawm Ca tsis txaus thiab cov ntshav Ca tsis ncav cuag qhov kawg, yuav muaj kev hloov pauv pathological xws li hypocalcemia thiab osteolysis, yog li kev noj zaub mov ntxiv ntawm Ca plays lub luag haujlwm tseem ceeb hauv kev tiv thaiv thiab txhim kho fluorosis (Dure-Smith et al., 1996; Li et al., 2021; Yang et al., 2021). Peb pawg tau lees paub tias kev noj hauscalciumyuav alleviate covapoptosisntawm pob txha thiab daim siab los ntawm PI3K-AKT teeb liab txoj kev, ER txoj kev, thiab mitochondrial txoj kev (Li et al., 2021; J. Wang et al., 2020; Wang et al., 2019; Yang et al., 2021). Yuav kom ua pov thawj ntxiv sebCalciumattenuates nephrotoxicity ntawm F los ntawm kev tuag receptor-mediatedapoptosistxoj hauv kev, peb tau tsim cov qauv nas ntawm cov khoom noj muaj fluoride siabcalciumthiab ntsuas qhovraumkev raug mob index, degree ntawmapoptosis, thiab cov kev hloov pauv ntawm cov cim noob caj noob ces thiab cov protein qhia nyob rau hauv qhov kev tuag receptor-mediatedapoptosistxoj kev.
Cov ntaub ntawv thiab cov txheej txheem
Tshuaj thiab cov cuab yeej siv
Cov dej distilled tau npaj los ntawm Heal Force dej purification System (Shanghai, Suav). Xov tooj cua immunoprecipitation assay (RIPA) lysis buffer thiab Sodium fluoride (NaF) tau muab los ntawm Sigma-Aldrich (Shanghai, Suav). 10 feem pua formalin, Glycine, sodium dodecyl sulfate (SDS), TRIS, Phenylmethylsulfonyl fluoride (PMSF), CaCO3, nitrocellulose (NC) membrane, Bicinchoninic Acid (BCA) Cov Khoom Siv thiab hematoxylin-eosin (H&E) staining kit tau txais los ntawm Solarbio Technology ., Ltd. (Beijing, Suav). Trizol reagent tau txais los ntawm Takara Biological Engineering Company (Dalian, Suav). CRE, UA, ntshav urea nitrogen (BUN), magnesium (Mg), P, thiab potassium (K) cov khoom siv tau yuav los ntawm Nanjing Jiancheng Bioengineering Institute (Nanjing, Suav). Hauv qhov chawapoptosisCov khoom siv kuaj pom, FAS Rabbit Polyclonal antibody (BA0484), thiab FAS-L Rabbit Polyclonal antibody (PB0042) tau yuav los ntawm Boster Biotechnology (Wuhan, Suav). -actin Luav Polyclonal antibody (20536-1-AP), FADD Rabbit Polyclonal antibody (14906-1-AP), Caspase 8 Rabbit Polyclonal antibody (13423-1-AP), Caspase 3 Rabbit Polyclonal antibody ({{9 }}AP), thiab HRPconjugated Affinipure Tshis Anti-Lavbit IgG(H plus L) (SA00001–2) tau txais los ntawm Proteintech Group (Wuhan, Suav). TRAL Rabbit Polyclonal Antibody (bs{13}}R), TNF alpha Rabbit Polyclonal antibody (bs2081R), PARP Rabbit Polyclonal Antibody (bs-55164R) tau yuav los ntawm Bioss (Beijing, Suav). Electrochemical luminescence (ECL) thiab 3,3′-diaminobenzidine (DAB) tau yuav los ntawm KeyGen Biotech (Jiangsu, Suav). PrimeScript® RT Master Mix thiab SYBR® Premix Ex Taq™ II Cov Khoom Siv tau muab los ntawm Promega Biotechnology (Beijing, Suav).
Tsiaj txhu thiab kev kho mob
Plaub caug noj qab nyob zoo {{0}}ib lub lis piam txiv neej Sprague-Dawley (SD) nas (120 ± 20 g) tau txais los ntawm Kev Tshawb Fawb Tsiaj Center ntawm Shanxi Medical University (Shanxi, Suav). Cov nas tau ua kom haum rau ib lub lim tiam, muab faib ua plaub pawg (10 nas rau ib pawg): Tswj pawg (C), NaF pawg (F), NaF ntxiv rau 0.5 feem pua CaCO3 pawg (F ntxiv 0.5 feem pua Ca), NaF ntxiv rau 1 feem pua. CaCO3 pawg (F ntxiv rau 1 feem puaCalcium) thiab kho raws li Table 1. Txhua tus nas tau nyob hauv lub tawb yas ntawm 22–25 ◦C (55 ± 5 feem pua ntawm cov av noo, 12 h lub teeb / tsaus voj voog), noj zaub mov thiab dej haus tau dawb.
Tom qab 17 lub lis piam ntawm kev yug me nyuam, nas yoo mov rau 24 teev, thiab dej tsis pub haus. Tom qab ntawd cov nas tsuag tau muab tshuaj loog nrog sodium pentobarbital thiab txi los ntawm lub ncauj tsev menyuam tom qab cov ntshav tau sau los ntawm lub qhov muag. Qee cov ntshav tau sau los ntawm cov hlab ntsha uas muaj cov tshuaj tiv thaiv heparin sodium rau kev kuaj BUN. Ib feem ntawm cov ntshav tau muab tso rau hauv cov raj sib xyaw, muab tso rau hauv chav sov li 2 teev, thiab tom qab ntawd centrifuged ntawm 3000 rpm / min rau 10 min kom cais cov ntshav. Cov ntshav tau muab cais rau CRE, UA, Mg, P, K, thiab F kev xeem. Yim tus qauv ntawmlub raumlos ntawm txhua pab pawg raug xaiv xaiv thiab khov sai sai hauv cov kua nitrogen, thiab tom qab ntawd khaws cia ntawm − 80 ◦C rau qRT-PCR thiab western blotting thwmsim. Lwm cov qauv ntawmlub raumtau kho nyob rau hauv 10 feem pua formalin rau histopathological kuaj thiab davhlau ya nyob twg deoxynucleotidyl transferase-mediated dUTP nick kawg labeling (TUNEL) thwmsim. Txhua qhov kev sim ua haujlwm txog tsiaj txhu tau ua nruj me ntsis raws li cov cai ntawm Lub Tsev Haujlwm Saib Xyuas tsiaj txhu thiab Pawg Neeg Siv Khoom ntawm Shanxi Agricultural University, Shanxi, Suav.
Lub raumcoefficient
Cov nas raug hnyav ua ntej; ces covraumcov ntaub so ntswg raug tshem tawm thiab hnyav. Covraumcoefficient tau txais los ntawm cov qauv hauv qab no.
Lub raumcoefficient=[raumntub hnyav (g) / lub cev hnyav (g)] × 100 feem pua.
Kev kuaj mob histopathological
Lub raumCov qauv tau muab tso rau hauv paraffin. Ntu (5 μm thickness) ntawm lubraumtau stained los ntawm H&E. Tom qab ntawd, cov slides raug soj ntsuam siv lub teeb pom kev zoo (Olympus, Tokyo, Nyiv). Qhov hnyav ntawm covraumKev raug mob raug txiav txim los ntawm cov qhab nia histopathological.Lub raumtubular raug mob yog txhais raws li tusraumcorpuscles raug atrophied thiab deformed, lubraumcapsule kab noj hniav widened, lubraumtubules tsim ib tug cam khwb cia, thiab epithelial hlwb ntog tawm. Ntawm 200 npaug ntawm kev nthuav dav, cov qauv hauv qab no tau siv los ntsuas qhov ntsuas ntawm daim siab raug mob. 1: 0-25% kev puas tsuaj; 2: 25–50 feem pua kev puas tsuaj; 3: 50–75 feem pua kev puas tsuaj; 4: 75–100 feem pua kev puas tsuaj (Baranova li al., 2016).
Kev txiav txim siab ntawmraum-Cov ntsuas ntsuas
Qib F yog ntsuas los ntawm F ion tshwj xeeb electrode (Quadri li al., 2018). Cov qib CRE, UA, BUN, Mg, P, thiab K raug ntsuas los ntawm kev siv cov khoom siv muag khoom. Tag nrho cov txheej txheem cuam tshuam tau ua raws li cov neeg tsim khoom cov lus qhia.
Kev kuaj pom ntawmapoptosistheem los ntawm TUNEL
Yuav kom quantify lubapoptosishauvraumcov hlwb, kev sim tshuaj ntsuam tau ua los ntawm kev siv qhov chawapoptosiskuaj cov khoom siv. CovraumCov seem tau deparaffinized thiab tom qab ntawd raug rau kev zom zaub mov los ntawm kev siv Proteinase K. Terminal Deoxynucleotidyl Transferase tuaj yeem khij digoxin-labeled dUTP (DIG-dUTP) mus rau 3′-OH kawg, thiab DIG-dUTP khi rau DNA breakpoint tom qab reacting nrog biomarker anti. - dig-Biotin. Ua ke nrog DAB tau ntxiv los tso saib. Rau visualizing
tag nrho cov nuclei, cov seem tau counterstained nrog hematoxylin. Tsaus xim av teeb liab qhia cov hlwb zoo thiab xiav qhia tias cov hlwb tsis teb. Covapoptosisindex tau xam raws li cov qauv hauv qab no.
Qhov feem pua ntawm apoptotic cells=(tus naj npawb ntawm apoptotic hlwb ntawm ib lub node / tag nrho cov xov tooj ntawm lub hlwb) × 100 feem pua.
Quantitative real-time PCR (qRT-PCR)
Total RNA los ntawmraumNws tau muab rho tawm los ntawm kev siv Trizol reagent. Peb siv ND{{0}} spectrophotometer (nano-poob, USA) thiab agarose gel electrophoresis los xyuas qhov zoo thiab muaj nuj nqis ntawm RNA. Reverse transcription (RT) tau ua tiav ntawm 500 ng tag nrho RNA los ntawm kev siv PrimeScript® RT Master Mix. Tshwj xeeb primers rau -actin thiab FAS, TRAIL, TNF, thiab lwm yam noob tau tsim los ntawm Primer Premier 7.0 software (Table 2) thiab synthesized los ntawm Sangon Biotech (Shanghai, Tuam Tshoj). Quantstudio 7 flex real-time PCR system (thermo-fisher, USA) thiab SYBR® Premix Ex Taq™ II Cov Khoom Siv tau siv rau RT-PCR. PCR cov tshuaj tiv thaiv ntim yog 20 μL, thiab RT-PCR cov xwm txheej yog: 95 ◦C 10
min, 40 lub voj voog ntawm 95 ◦C 15 s, 55 ◦C 30 s, 72 ◦C 30 s, thiab thaum kawg ib lub voj voog ntawm 95 ◦C 15 s, 60 ◦C 15 s, 95 ◦C 15 s. Txhua qhov kev tshwm sim tau rov ua dua peb zaug. Tus txheeb ze 2-ΔΔCt txoj kev tau siv los xam cov txheeb ze mRNA qhia qib, thiab -actin tau siv los ua tus ntsuas ntsuas. Hauv cov txheej txheem, qhov sib txawv ntawm qhov ua tau zoo yog tsawg dua 5 feem pua.
Tag nrho cov protein extraction thiab western blotting tsom xam
Covraumcov ntaub so ntswg tau ntxuav nrog PBS, qhuav nrog cov ntawv lim, thiab decomposed hauv RIPA lysis (muaj PMSF), ntawm 12,000 rpm/min, 4◦C rau 10 min. Protein concentration tau txiav txim los ntawm kev siv cov khoom siv Bicinchoninic Acid (BCA) (Solarbio Science & Technology, Beijing, Suav). 35 ng protein cov qauv tau ua tiav zoo thiab sib cais ntawm 8 feem pua SDS-polyacrylamide gel los ntawm electrophoresis. Lub hom phiaj protein tau pauv mus rau NC daim nyias nyias, thiab NC daim nyias nyias tau thaiv nrog 5 feem pua ntawm cov mis nyuj uas tsis muaj rog nyob hauv chav sov li 2 teev. Tom qab ntawd, NC membrane tau incubated nrog thawj cov tshuaj tiv thaiv ntawm -actin (1: 1000), TRAIL (1: 500), FAS (1: 500), FAS-L (1: 100), TNF (1: 500), FADD (1:500), Caspase 8 (1:500), Caspase 3 (1:500) thiab PARP (1:1000) ntawm 4 degree thaum hmo ntuj. Tom qab ntxuav 3 zaug nrog PBST, NC daim nyias nyias tau tsim nrog HRP-conjugated theem nrab tshis tiv thaiv tus luav IgG cov tshuaj tiv thaiv (1: 2000) ntawm chav tsev kub rau 2 teev. ECL tau siv los ua kom pom lub hom phiaj protein bands. Optical density yog xam los ntawm AlphaView software (version: 3.2.2.0) ntawm FluorChem Q system (Alpha Innotech, CA, USA).
Kev txheeb cais
GraphPadPrism{{0}} software tau siv los teeb tsa thiab txheeb xyuas cov ntaub ntawv sim, thiab txhua cov ntaub ntawv tau sawv cev los ntawm txhais tau tias ± SEM. Qhov tseem ceeb ntawm qhov sib txawv ntawm txhais tau tias yog txiav txim los ntawm kev tsom xam ntawm qhov sib txawv (ANOVA) ua raws li Tukey qhov kev sim nrog P <0.05 tau="" suav="" tias="" yog="" qhov="" tseem="">
Cov txiaj ntsig
Kev hloov pauv hauvraumCoefficient thiab biochemical muaj nuj nqi ntsuas
Covraumcoefficient thiab theem ntawm F, CRE, BUN, UA, Mg, P, K tau qhia nyob rau hauv daim duab 1.raumcoefficient tau txo qis hauv pawg F (p < 0.05).="" txawm="" li="" cas="" los="" xij,="" lub="" raum="" coefficient="" hauv="" f="" ntxiv="" rau="" 1="" feem="">Calciumpab pawg tau pom qhov nce ntxiv raws li piv nrog F pawg (p < {{0}}}05).="" qib="" f="" hauv="" pawg="" f="" yog="" qhov="" siab="" tshaj="" hauv="" pawg="" c="" (p=""><0.001). interestingly,="" piv="" rau="" f="" pawg,="" f="" qib="" hauv="" f="" ntxiv="" rau="" 1="" feem="" pua="" ca="" pab="" pawg="" tau="" poob="" qis="" (p=""><>
CRE, BUN, thiab UA tau raug tshuaj xyuas los cuam tshuam qhov degree ntawmraumkev puas tsuaj thiab kev hloov pauv. Cov theem ntawm CRE, BUN, thiab UA tau nce ntxiv hauv pawg F piv rau cov hauv pawg C (p < 0.05),="" txawm="" li="" cas="" los="" xij,="" txheeb="" ze="" rau="" pawg="" f,="" peb="" qhov="" ntsuas="" saum="" toj="" no="" yog="" qhov="" tseem="" ceeb.="" txo="" qis="" hauv="" f="" ntxiv="" rau="" 1="" feem="">Calciumpab pawg (p < 0.05).="" mg,="" p="" thiab="" k="" yog="" cov="" ntsuas="" electrolyte="" ze="" ze="">raummuaj nuj nqi. Piv nrog rau pawg C, qib ntawm cov ntshav Mg tau txo qis, cov qib P thiab K tau nce siab hauv pawg F (p < 0.05).="" txawm="" li="" cas="" los="" xij,="" ntxiv="" 1="" feem="">calciumrau kev noj zaub mov tshwj xeeb tau nce qib Mg hauv cov ntshav los ntawm 15.3 feem pua (p < 0.05)="" thiab="" txo="" cov="" ntshav="" qab="" zib="" ntau="" ntau="" los="" ntawm="" 16.9="" feem="" pua="" (p=""><0.05). cov="" ntsiab="" lus="" ntawm="" cov="" ntshav="" k="" tau="" pom="" qhov="" poob="" qis="" uas="" tsis="" muaj="" qhov="" tseem="" ceeb="" tom="" qab="" ntxiv="" 1="" feem="" pua="" ca="">
Hloov hauvraummorphology
Cov kev hloov morphological ntawmraumCov ntaub so ntswg raug tshuaj xyuas los ntawm H&E (Fig. 2I-Ⅳ, ⅰ-ⅳ). Hauv pawg C,raumTubular hlwb tau compactly teem, lub morphology yog tsis tu ncua, thiab morphology ntawm glomeruli yog lawm. Txawm li cas los xij, cov ciam teb ntawm cov hlwb tsis meej, lubraumcorpuscles tau atrophied thiab deformed, lubraumcapsule kab noj hniav widened, lubraumtubules tsim ib tug cam khwb cia, thiab epithelial hlwb tau detached nyob rau hauv pawg F. Nyob rau hauv kev sib piv rau pawg F, nrog rau kev nce ntawmCalciumconcentration, kev npaj ntawmraumcov hlwb maj mam ua kom zoo dua, lub morphology ntawmraumcorpuscles maj mam zoo, thiab cov degree ntawm kev raug mob yog maj alleviated. Tsis tas li ntawd, peb tau soj ntsuam qhov kev puas tsuaj los ntawm kev siv lubraumtau qhab nia (Fig. 2Ⅴ). Cov qhab nia ntawm pawg F tau nce siab dua li ntawm pawg C (p < 0.01);="" txawm="" li="" cas="" los="" xij,="" qhov="" qhab="" nia="" hauv="" f="" ntxiv="" rau="" 1="" feem="">Calciumpab pawg poob qis heev thaum piv rau pawg F (p < 0.05).
TUNEL kuaj pomraumcellapoptosis
Peb tau ua qhov kev ntsuam xyuas TUNEL txhawm rau txheeb xyuas thiab txheeb xyuas seb puas yograumcov hlwb tau dhau lawmapoptosisnyob rau hauv txoj kev tshawb no (Fig. 3I-Ⅳ, ⅰ-ⅳ).Lub raumCov ntaub so ntswg hauv C pab pawg tau raug kho kom zoo, nrog tsawg dua apoptotic hlwb (4.67 ± 1.15 feem pua). Pawg F tau pom ntau tus TUNEL-zooraumtubular epithelial hlwb (48.17 ± 3.94 feem pua). Hauv qhov sib piv, F ntxiv rau 1 feem puaCalciumpab pawg tau pom qhov txo qis ntawm TUNEL-zoo hlwb (28.83 ± 4.81 feem pua) (Daim duab 3Ⅴ).
Cov teebmeem ntawmCalciumntawm F-induced alterations nyob rau hauv kev tuag receptor pathway-related genes
Cov txheeb ze mRNA qhia theem ntawm cov dej ntws ntws muaj feem cuam tshuam nrog kev tuag receptor-mediatedapoptosistxoj kev (TRAIL, DR5, TNF, TNFR1, TNF-R2, FAS, FAS-L, OPG) tau tshwm sim hauv daim duab 4a. Kev txheeb ze rau pawg C, mRNA qhia theem ntawm TRAIL, DR5, TNF, TNF-R1, TNFR2, FAS, thiab FAS-L tau tshwj xeeb hauv F-kho nas (p < 0.01).="" hauv="" qhov="" sib="" piv,="" cov="" txheeb="" ze="" mrna="" kab="" lus="" ntawm="" trail,="" tnf,="" tnf-r2,="" fas,="" thiab="" fas-l="" tau="" txo="" qis="" hauv="" f="" ntxiv="" rau="" 1="" feem="">Calciumpab pawg raws li piv nrog pawg F (p < {{0}}.05).="" hauv="" kev="" sib="" piv="" rau="" pawg="" c,="" qhov="" pom="" tseeb="" txo="" qis="" hauv="" mrna="" qhia="" theem="" ntawm="" opg="" hauv="" pawg="" f="" tau="" pom="" (p=""><0.01). txawm="" li="" cas="" los="" xij,="" txheeb="" ze="" rau="" pawg="" f,="" opg="" hauv="" f="">Calciumpawg tau nce. Cov xwm txheej hauv qab FADD, TRADD, Caspase 8, Caspase 3, thiab PARP txheeb ze mRNA qhia qib tau qhia hauv daim duab 4b. Cov lus qhia protein ntawm FADD, TRADD, Caspase 8, Caspase 3, thiab PARP tau pom tseeb txhim kho hauv pawg F raws li piv nrog pawg C (p < 0.01),="" tab="" sis="" pom="" tseeb="" txo="" qis="" hauv="" f="">Calciumpab pawg, piv nrog rau pawg F (p < 0.05).
Cov teebmeem ntawmCalciumntawm F-induced alterations nyob rau hauvapoptosis- muaj feem xyuam rau cov protein
Cov protein qhia ntawm TRAIL, FAS, FAS-L, TNF, FADD, Caspase 8, Caspase 3, thiab PARP tau kuaj pom los ntawm Western blotting (Fig. 5a, b). Piv nrog rau pawg C, cov lus qhia protein ntawm TRAIL, FAS, FAS-L, TNF, FADD, Caspase 8, Caspase 3, thiab PARP hauv pawg F tau nce siab heev (p < 0.01).="" cov="" kev="" qhia="" ntawm="" cov="" protein="" ntawm="" tag="" nrho="" cov="" saum="" toj="" no="" tseem="" ceeb="" txo="" qis="" hauv="" f="" ntxiv="" rau="" 1="" feem="">Calciumpab pawg thaum piv rau pawg F (p < 0.05).
Kev sib tham
Cov pov thawj tsis ntev los no tau ua pov thawj tias F tuaj yeem ua rau cov nqaij mos mos, thiab qhov kev puas tsuaj ntawm F yog nyob ntawm qhov concentration ntawm F, lub sijhawm raug, thiab hom kabmob (Wei li al., 2019). Hauv txoj kev tshawb no, 100 mg / L NaF tau siv rau 17 lub lis piam los tsim kom muaj cov dej haus subacute NaF raug tus qauv tsiaj. Qhov ntau npaum ntawm NaF raug xaiv raws li cov pa phem ntawm F hauv ib puag ncig thiab qhov tsis paub tseeb ntawm ntau hom. Muab hais tias lub ntuj F- concentration ntsuas nyob rau hauv cov av hauv av yog kwv yees li 4.5 mg / L, cov dej distilled nrog 100 mg / L NaF siv nyob rau hauv txoj kev tshawb no (qhov F- concentration yog li ntawm 45 mg / L) raug xam raws li qhov tsis paub tseeb. tsiaj-rau-tib neeg extrapolation ntawm 10 (Cov Lus Qhia rau Kev Haus Dej Zoo, 2017; Mukherjee thiab Singh, 2021; Wen et al., 2013; Zhang et al., 2020). Cov kev tshawb fawb thaum ntxov tau pom tias kev noj hauscalciumtuaj yeem cuam tshuam rau kev nqus ntawm fluorine, txhawb kev tso tawm ntawm fluorine hauv lub cev, txo qhov nqus ntawm cov tshuaj fluorine, thiab yog li txo cov toxicity ntawm F (Harrison li al., 1984; Larsen li al., 1981). Peb cov kev tshawb fawb tsis ntev los no tau pom tias Ca alleviates F-induced pob txha thiab lub siab raug mob los ntawm inhibiting txoj hauv kev ntawmapoptosis(Li et al., 2021; J. Wang et al., 2020; Wang et al., 2019). Txawm li cas los xij, txog tam sim no, kev tshawb fawb txog kev tiv thaiv ntawm Ca ntawm F-induced nephrotoxicity thiab nws cov txheej txheem cuam tshuam tsis tau muaj dua. Ntawm no, peb tshaj tawm tias Ca alleviates F-inducedraumkev puas tsuaj. Tsis tas li ntawd, peb kuj pom tias Ca thim rov qab F-inducedraumcellapoptosislos ntawm kev tuag receptor-mediatedapoptosisTxoj kev (FAS/FASL, TNFR/TNF, DR5/TRAIL pathways). Lub sij hawm ntev tsub zuj zuj ntawm F nyob rau hauv lubraumtuaj yeem ua rau cov ntaub so ntswg thiab kev ua haujlwm puas. Hauv txoj kev tshawb no, peb pom tias NaF raug tshwm simraumcellapoptosisnyob rau hauv nas, nrog rau glomerular atrophy thiab deformation,raumcapsule kab noj hniav widening,raumtubules tsim casts, thiab cov hlwb epithelial ntog tawm. Txawm li cas los xij,raumKev puas tsuaj tau txo qis tom qab ntxiv ntawm 1 feem pua Ca. Cov txiaj ntsig no zoo ib yam li cov txiaj ntsig tau txais hauv cov ntaub ntawv dhau los (Cao et al., 2015; HW Wang et al., 2020).
CRE yog cov metabolite ntawm cov leeg, UA yog cov khoom kawg ntawm purine metabolism, thiab BUN yog cov khoom tseem ceeb kawg ntawm lub cev cov protein metabolism, uas yog tawm los ntawm glomerular pom thiab feem ntau yog siv los kuaj.raummuaj nuj nqi (Myers et al., 2006; HW Wang et al., 2020). Kev tshawb fawb biochemical ntawm thaj tsam F siab tau pom tias cov glomerular pom tus nqi tau txo qis, thiab F, UA, thiab BUN hloov pauv ntau (Malin li al., 2019). Cov tsiaj kuaj pom tias cov ntshav CRE,Calcium, thiab P concentrations tau txo qis hauv cov nas uas raug rau 100 mg / L NaF, qhia tias F cuam tshuamraumKev ua haujlwm zoo li Ca thiab P metabolism (HW Wang li al., 2020). F-induced nephrotoxicity muaj feem xyuam nrograumkev ua haujlwm tsis zoo. Thaum twgraumkev ua haujlwm tsis zoo, qhov tso tawm ntawm F hauv lub cev raug txo, ua rau ntau dhau ntawm cov tshuaj fluorine hauv lub cev.raumthiab aggravating F toxicity (Kido li al., 2017). Thaum lublub raumraug mob hnyav heev, cov
Kev tso tawm ntawm F hauv cov zis txo qis, thiab ntshav F concentration ntxiv ntxiv (Dharmaratne, 2019). Hauv txoj kev tshawb no, peb tau pom tias cov qib BUN thiab cov ntshav CRE, UA, P, K hauv cov nas uas kho nrog 100 mg / L NaF tau nce ntau, qhia tias qhov F raug siab ua rau muaj kev hloov pauv hauvraumbiochemical ntsuas hauv nas thiab coj mus rauraumtsis txaus. Tsis tas li ntawd, peb kuj tau taw qhia tias ntau yam ntsuas ntawm lubraumzoo li qub tom qabCalciumkev pab ntxiv. Nws qhia tau tias Ca (tshwj xeeb tshaj yog 1 feem puaCalcium) muaj qhov cuam tshuam tseem ceeb ntawm F-inducedraumkev puas tsuaj hauv nas.
Cistanchetuaj yeem pab nrograumkev puas tsuaj.
Apoptosisyog ib tug autonomous thiab orderly cell tuag tswj los ntawm noob, uas yog yus muaj los ntawm DNA degradation tsis pom tseeb cell lysis. Txoj kev tshawb no pom tias fluorosis tshwm simraumcellapoptosishauv nas. FAS plays lub luag haujlwm tseem ceeb hauv qhov tshwm sim ntawmapoptosisthiab ntau yam kab mob. Cov ntaub ntawv tsis ntev los no tau tshaj tawm tias FAS sib thamapoptosishauvrauminduced los ntawm ischemia-reperfusion thiab tib neeg leukocytesapoptosisinduced los ntawm N-nitrosodimethylamine (Iwaniuk li al., 2019; Xu
et al., 2019). Thaum FAS thiab FAS-L ua ke los tsim ib lub trimer, pro-apoptotic teeb liab tau tshwm sim. Kev tshawb fawb tsis ntev los no tau qhia tias F induces cellapoptosislos ntawm txoj kev FAS/FAS-L (Xu et al., 2011). Hauv qhov kev tshawb fawb no, F tau nce ntau ntxiv ntawm mRNA thiab cov protein qhia ntawm FAS thiab FAS-L hauvraum. Kev tswj hwm ntawm FAS thiab FAS-L qib qhia tias FAS / FAS-L-txog txoj hauv kev apoptotic tuaj yeem koom nrog F-inducedapoptosis. FAS / FAS-L trimer recruitment ntawm FADD ua rau lub cleavage ntawm Pro-Caspase 8 los tsim active Caspase 8, uas nyob rau hauv lem activates Caspase 3 thiab ces coj mus rauapoptosis(Li et al., 2011; Xu et al., 2011). Txoj kev tshawb no tau txheeb xyuas tias 100 mg / L NaF inducedapoptosishauv ratlub raumlos ntawm txoj kev FAS/FAS-L. Qhov tseem ceeb tshaj plaws yog tias kev noj haus Ca supplementation attenuates lubapoptosis-inducing nyhuv ntawm F, uas yog ntaus nqi rau down-regulation ntawm FAS / FAS-L ua kom thiab theem nrab ua kom Caspases.
TNF yog pleiotropic cytokine uas koom nrog ntau cov lus teb ntawm tes, suav nrog kev sib txawv, kev loj hlob, kev mob, thiab kev tuag ntawm tes. TNF teeb liab ua rau ob qho tib siapoptosisthiab anti-apoptotic txoj kev. Ntau hom ntaub ntawv tau tshaj tawm tias TNF dominates lipopolysaccharide-inducedraumkev puas tsuaj thiab cisplatin-induced mobraumkev raug mob (Lee et al., 2016; Li et al., 2018). TNF khi rau TNF-R1, nws DD nrhiav TRADD, thiab tom qab ntawd TRADD cuam tshuam nrog FADD, ua rau muaj kev tsim DISC (Kiraz li al., 2016; Sun et al., 2003). Cov kev tshawb fawb tau qhia tias F tuaj yeem ua rau lub siab, neuron, thiabraumleukocyteapoptosislos ntawm TNF signaling pathway (Lu et al., 2017; Singh et al., 2017; Yan et al., 2016). Zoo ib yam li cov txiaj ntsig ntawm cov ntawv ceeb toom no, cov lus txheeb ze ntawm TNF, TNF-R1, TNF-R2, thiab TRADD hauv pawg F tau nce ntau hauv qhov kev sim no. Los ntawm qhov sib txawv, qhov kev qhia ntawm cov noob no poob qis tom qab 1 feem puaCalciumntxiv, qhia txog kev cuam tshuam ntawmCalciumhauv TNF txoj kev.
TRAIL yog koom nrog hauv txoj cai ntawmapoptosis, proliferation, immune-inflammatory teb, thiab lwm yam. Tam sim no, nws ntseeg hais tias TRAIL yog ze ze rau qhov tshwm sim thiab prognosis ntawmraumkab mob (Candido, 2014). Ntau thiab ntau cov ntaub ntawv tau lees paub tias TRAIL tswj covapoptosisntawmraumtubular epithelial hlwb hauv HBV-koom nrog glomerulonephritis thiabraumapoptosisnyob rau hauv Uromodulin-koom nrograumkab mob (Johnson et al., 2017; Yang et al., 2018). Tsis tas li ntawd, ib txoj kev tshawb fawb tau tshaj tawm tias qhov kev qhia ntawm TRAIL feem ntau hloov pauv thaum lub sij hawm homeostasis thiab ntau yamraumkev raug mob (Devarapu li al., 2017). TRAIL txuas nrog tus neeg txais kev tuag DR5 thiab nrhiav FADD los ntawm kev sib cuam tshuam ntawm DD, thiab tom qab ntawd khi rau pro-Caspase 8 los ntawm kev tuag cov txiaj ntsig muaj nyob hauv FADD los tsim DISC (Yuan li al., 2018). Txoj kev tshawb no tau pom tias qhov kev qhia ntawm TRAIL thiab DR5 tau nce ntau hauv pawg F. Tom qab supplementing nrogCalcium, kev qhia ntawm TRAIL thiab DR5 raug txwv. Yog xav tiasCalciuminhibits txoj kev TRAIL kom txo F-inducedapoptosis.
Xaus
100 mg/L NaF raug activated death receptor-mediatedapoptosistxoj kev (FAS/FASL, TNFR/TNF, DR5/TRAIL txoj kev) thiab ces inducedraumcellapoptosishauv nas, ua rauraummetabolic ntshawv siab thiabrauminsufficiency, ua rau covraumcorpuscles tau atrophied thiab deformed, lubraumcapsule kab noj hniav widened, lubraumtubules tsim ib tug cam khwb cia. Txawm li cas los xij, qhov sib ntxiv ntawm 1 feem puaCalciumrau kev noj haus txo cov ntshav fluoride cov ntsiab lus, ntxiv alleviated lubraummetabolic ntshawv siab thiabrauminsufficiency los ntawm txoj kev tuag receptor-mediated txoj kev, thiab ho txo tusraumkev raug mob (tej daim duab 6).
Cov ntaub ntawv
Lub hauv paus yog los ntawm Haojie Li, College of Veterinary Medicine, Shanxi Agricultural University, Taigu 030801, Shanxi, PR Tuam Tshoj thiab lwm yam.
