Keeb kwm.Mob hnyavmob raum(AKI) yog ib qho teeb meem ntau zaus ntawm decompensated cirrhosis nrog kev tuag ntau ntxiv. Ib txwm biomarkers xws li ntshav creatinine tsis nkag siab rau kev kuaj pom kev raug mob yam tsis muaj kev hloov pauv. Peb xav tias cov zis exosomes muaj peev xwm nqa cov cim uas txawv hom ntawmmob raumhauv cov neeg mob cirrhotic.

Cov txheej txheem.Qhov no yog qhov kev cia siab, ib qho chaw, thiab kev soj ntsuam ntawm cov neeg laus uas muaj mob cirrhosis. Cov pab pawg neeg mob suav nrog kev tswj hwm kev noj qab haus huv, them nyiaj cirrhosis nrog qhov qublub raum ua haujlwm, decompensated cirrhosis nrog ib txwmlub raum ua haujlwm, thiab decompensated cirrhosis nrog AKI. Cov ntaub ntawv raug muab rho tawm los ntawm cov ntaub ntawv kho mob hauv hluav taws xob suav nrog etiology ntawm kab mob siab, MELD tus qhab nia, keeb kwm ntawm decompensation, Child-Turcotte-Pugh qhab nia, keeb kwm ntawm AKI, thiab kev siv tshuaj. Cov zis kuaj tau sau thaum lub sijhawm tso cai. Cov zis exosome protein cov ntsiab lus tau txheeb xyuas, thiab cov ntaub ntawv proteomic tau raug lees paub los ntawm immunoblotting. Kev txheeb xyuas txheeb cais suav nrog ib feem tsawg kawg squares-kev cais cais tawm ua ke nrog qhov tseem ceeb sib txawv hauv kev txheeb xyuas qhov projection.

Cov txiaj ntsig.Kaum yim tus neeg mob cirrhotic tau cuv npe, thiab rau qhov kev tswj hwm kev noj qab haus huv tau muab rho tawm los ntawm peb lub biorepository. Cov zis exosomes raug cais tawm, thiab 1572 cov proteins raug txheeb xyuas. Maltase-glucoamylase yog cov tshuaj tiv thaiv kab mob siab tshaj plaws uas tau lees paub los ntawm western blotting.

Cov lus xaus. Cov neeg mob uas muaj cirrhosis thiab AKI muaj upregulation ntawm lub raum txhuam ciam teb disaccharidase, MGAM, nyob rau hauv urinary exosomes uas tej zaum yuav txawv hom.mob raumnyob rau hauv cirrhosis; Txawm li cas los xij, qhov tseem ceeb ntawm kev kho mob ntawm qhov no yuav tsum muaj kev lees paub ntxiv.

Yog xav paub ntxiv thov hu rau:joanna.jia@wecistanche.com

Mob raum mobkho los ntawmcistanche

1. Taw qhia

Mob hnyavmob raum(AKI) tshwm sim nyob rau hauv kwv yees li 20 feem pua ​​​​ntawm cov neeg mob nyob hauv tsev kho mob uas muaj cirrhosis [1, 2]. AKI nyob rau hauv tsev kho mob cirrhotic cov neeg mob feem ntau nce zuj zus, hnyav, thiab muaj kev ywj pheej tsis zoo rau kev tuag [3]. *e feem ntau ua rau AKI hauv cirrhosis yog hemodynamic, suav txog 70 feem pua ​​​​ntawm cov neeg mob. Mob tubular necrosis (ATN) suav txog 30 feem pua ​​​​ntawm cov neeg mob, thiab cov tsos mob tom qab tsis tshua muaj nyiaj tsawg dua 1 feem pua ​​​​ntawm cov neeg mob. Hepatorenal syndrome (HRS) yog hemodynamic yam tsis muaj kev txheeb xyuasmob raumlos yog kab mob thiab tshwm sim nyob rau hauv kwv yees li 20 feem pua ​​​​ntawm cov neeg mob cirrhosis [4, 5].

Serum creatinine (Scr) yog siv dav siv biomarker los ntsuaslub raum ua haujlwmthiab txheeb xyuasmob raum. Txawm li cas los xij, SCR yog qhov zoo tshaj plaws hauv cov neeg mob cirrhotic rau Hindawi Critical Care Research thiab Practice Volume 2019, ntau lub laj thawj suav nrog kev txo qis hauv siab, cov nqaij ntshiv nrog cov khw muag khoom poob qis, nce ntim ntawm kev faib tawm, thiab protein-calorie malnutrition. Qhov nruab nrab Scr qhov tseem ceeb yog qis dua hauv cov neeg mob cirrhotic piv rau cov pej xeem, ua rau ncua kev kuaj mob AKI raws li cov ntsiab lus tam sim no ntawm AKI [6]. Tsis tas li ntawd, Scr yog biomarker ntawmlub raum ua haujlwmthiab tsis yog ib qho kev raug mob rhiab heev. Novel biomarkers ntawm lub raum raug mob tau tshwm sim los txhim kho kev tshawb pom ntawm AKI thiab pab hauv kev sib txawv ntawm etiology ntawm AKI.Lub raum puasbiomarkers suav nrogmob raummolecule -1 (KIM-1), neutrophil gelatinase-associated lipocalin (NGAL), interleukin-18, daim siab fatty acid-binding protein (L-FABP), insulin-zoo li kev loj hlob zoo li kev khi cov protein -7 (IGFBP-7), thiab cov ntaub so ntswg inhibitor ntawm metalloproteinase-2 (TIMP-2) tuaj yeem nce siab ua ntej kev nce hauv Scr txhim kho kev tshawb pom ntawmraum puasyam tsis muaj kev hloov pauv [7]. Cov kev tshawb fawb tau pom tias cov biomarkers no yuav txawv qhov etiology ntawm AKI [8, 9]. Txhim kho biomarkers rau kev kuaj pom thiab sib txawv AKI sawv cev tseem ceeb hauv kev xav tau kev kho mob tsis zoo hauv cov neeg mob cirrhosis.

Exosomes yog nanovesicles uas tau tso tawm los ntawm cov hlwb nyob ua ib qho kev sib txuas lus ntawm intercellular [10]. *e protein cov ntsiab lus ntawm exosomes tau pom tias tau hloov kho zoo kawg nkaus nyob rau hauv pathological lossis kev ntxhov siab [11–13]. Hauvraum,exosomes raug xa mus rau cov zis los ntawm txhua hom cell [14, 15], thiab tso zis exosomes tuaj yeem suav hais tias yog qhov kos npe biochemical ntawm qhov kev kawm. Txij li cov exosomes urinary tsis niaj hnub soj ntsuam, lawv tuaj yeem muab cov ntaub ntawv ntxiv tsis paub txog cov protein biomarkers ntawm AKI hauv cov neeg mob cirrhosis.

Lub hom phiaj ntawm txoj kev tshawb no yog los soj ntsuam cov urinary exosome proteomics hauv cov neeg mob cirrhosis thiab AKI piv rau cov kev noj qab haus huv. Peb tau xav tias cov ntsiab lus ntawm cov zis exosome protein txawv nyob rau hauv cov neeg mob uas tau them nyiaj los yog decompensated cirrhosis muaj AKI piv rau cov kev tswj kev noj qab haus huv. Tsis tas li ntawd, peb tau tshaj tawm tias qhov sib txawv ntawm cov zis exosomal protein cov ntsiab lus yuav muab kev pom rau cov txheej txheem ntawmmob raumhauv cirrhosis.

2. Cov ntaub ntawv thiab cov txheej txheem

Qhov no yog qhov kev cia siab, ib qho chaw, thiab kev soj ntsuam ntawm cov neeg laus uas muaj mob cirrhosis. Txhua tus neeg mob rau txoj kev tshawb fawb tau raug xaiv los ntawm UC San Diego Health system thaum Lub Xya Hli 1, 2013, thiab Lub Rau Hli 1, 2014, thiab tau txais kev pom zoo. Cov neeg mob tau tsim nyog suav nrog yog tias lawv tau kuaj pom tus mob cirrhosis thiab muaj peev xwm muab cov zis tso zis. Cirrhosis tau txiav txim siab los ntawm daim siab biopsy, hla ntu duab, lossis kev kho mob (los ntawm kev txheeb xyuas qhov kev tshwm sim decompensation raws li kev txiav txim siab los ntawm kws kho mob hepatologist). Cov ntaub ntawv raug muab rho tawm los ntawm cov ntaub ntawv kho mob hauv hluav taws xob suav nrog cov pej xeem, cov kab mob anthropometrics, cov cim tseem ceeb, cov teeb meem kho mob comorbid, etiology ntawm cirrhosis, cov teeb meem ntawm cirrhosis (ascites, varices, hepatic encephalopathy, thiab hepatocellular carcinoma), keeb kwm ntawm AKI los yogmob raum mob, thiab raug tshuaj nyob rau hauv 30 hnub ntawm kev tso npe. Tsuas yog cov neeg mob uas ua tiav cov ntaub ntawv kho mob thiab kev sim kuaj hauv 30 hnub ntawm kev tso npe kawm tau tsim nyog rau kev suav nrog hauv txoj kev tshawb no. Cov neeg mob tau muab faib ua pawg raws li hauv qab no:

(1) Pawg 0: tswj kev noj qab haus huv

(2) Pawg 1: Kev them nyiaj yug cirrhosis (Child-Turcotte- Pugh chav A, MELD 10) tsis muaj keeb kwm ntawm AKI thiab ib txwm muajlub raum ua haujlwm

(3) Pawg 2: decompensated cirrhosis (Child-Turcotte- Pugh chav B lossis C) uas tsis muaj keeb kwm ntawm AKI thiab ib txwm muajlub raum ua haujlwm

(4) Pawg 3: decompensated cirrhosis (Child-Turcotte- Pugh chav B lossis C) thiab AKI

Ib txwmlub raum ua haujlwmtau txhais tias yog kwv yees li GFR > 60 ml/min/1.73 m2 (MDRD formula), tsis muaj albuminuria, thiab tsis muaj keeb kwm ntawm AKI. AKI tau txhais raws li AKIN cov txheej txheem: Scr nce ntawm 0.3 mg / dl hauv 48 hr lossis 50 feem pua ​​​​nce hauv Scr los ntawm lub hauv paus [16]. Cov neeg mob nrog AKI tau raug xaiv los ntawm kev nkag mus thiab kev sab laj los ntawm qhov chaw kho mob hepatology yog tias lawv muaj cov ntshav thiab cov zis tau txais thaum lub sijhawm AKI. Pawg plaub ntawm kev tswj hwm kev noj qab haus huv tau muab rho tawm los ntawm kev noj qab haus huv biorepository ntawm UCSD O'Brien Center rau AKI Kev Tshawb Fawb ntawm UC San Diego Tsev Kawm Ntawv Tshuaj. * yog txoj haujlwm tau pom zoo los ntawm Pawg Saib Xyuas Kev Tshawb Fawb ntawm University of California, San Diego.

2.1. Kev kuaj zis thiab ua rau Exosome cais tawm.

Cov zis tau centrifuged ntawm 3000 ×g rau 30 feeb. Lub supernatant pH tau hloov mus rau 7, aliquoted, thiab khov ntawm -80 degree C. Exosomes tau npaj siv polyethylene glycol- (PEG-) induced nag lossis daus [17]. *e PEG-cov zis kuaj tau muab tso rau hauv chav sov li 2 teev thiab spun ntawm 10,000 ×g rau 30 feeb. * e pellet tau resuspended hauv 10 mM Tris nrog 1 mM EDTA-Na ntsev. * yog kauj ruam tau rov ua dua ob zaug kom tshem tawm impurities. Ib-dimensional SDS PAGE ntawm exosome proteins tau ua ua ntej nyob rau hauv-gel trypsinization los tiv thaiv confounding [18].

2.2. Proteomic Analysis.

Cov gel raug txiav mus rau 1 mm × 1 mm thiab detained 3 zaug siv 100 µL ntawm 100 mM ammonium bicarbonate rau 15 feeb, tom qab ntawd 100 µL ntawm acetonitrile (ACN) rau 15 feeb [19 ]. *e supernatant tau lyophilized, thiab qhov tshwm sim pellet raug txo nrog 200 µL ntawm 100 mM ammonium bicarbonate-10 mM DTT thiab incubated ntawm 56 degree rau 30 feeb. Tom qab tshem tawm cov kua, cov gels tau ntxiv rau 200 µL ntawm 100 mM ammonium bicarbonate -55 mM iodoacetamide. * yog incubated ntawm chav tsev kub rau 20 feeb nyob rau hauv qhov tsaus ntuj nti. * e supernatant raug tshem tawm thiab ntxuav nrog 100 mM ammonium bicarbonate rau 15 feeb. * en, 100 µL ntawm ACN tau ntxiv rau lub cev qhuav dej ntawm cov gel, thiab cov tshuaj tau lyophilized. Ice-txias trypsin (0.01 µg / µL) hauv 50 mM ammonium bicarbonate tov tau ntxiv los npog cov gel daim rau cov txheej txheem zom thiab teem rau dej khov rau 30 feeb. Thaum rov ua dej tiav lawm, cov tshiab 50 mM ammonium bicarbonate tau ntxiv los hloov qhov ntau dhau. trypsin thiab sab laug hmo ntuj ntawm 37 degree. Extraction ntawm peptides tau ua ob zaug los ntawm kev sib ntxiv ntawm 50 µl ntawm 0.2 feem pua ​​formic acid thiab 5 feem pua ​​ACN thiab vortexed rau 30 feeb ntawm chav tsev kub. Tom qab tshem tawm cov supernatant, 50 µl ntawm 50 feem pua ​​ACN -0.2 feem pua ​​formic acid tau ntxiv rau cov qauv, vortexed dua rau 30 feeb ntawm chav tsev kub. * yog supernatant raug tshem tawm thiab ua ke nrog yav dhau los supernatant los ntawm thawj rho tawm. Cov qauv raug txheeb xyuas siv Eksigent nano-LC-Ultra® 2D nrog cHiPLC-nano flex system (Eksigent, AB SCIEX Dublin, CA, USA) hauv hom ntxiab-elute ua ke nrog tandem huab hwm coj spectroscopy siv QExactive huab hwm coj spectrometer (Thermo Fisher Scientific, San Jose', CA, USA) nrog electrospray ionization [17].

2.3. Kev Tswj Cov Ntaub Ntawv.

Lub SEQUEST search engine (Thermo Scientific Proteome Discoverer software, version 1.4) tau siv rau hauv kev tshuaj xyuas. *e protein database rau tryptic peptide sequences rau Homo sapiens los ntawm National Center for Biotechnology Information (NCBI) tau siv los sib piv peb qhov kev sim MS/MS spectra. Txhawm rau txheeb xyuas peptide ib ntus thiab cov proteins uas muaj feem xyuam, peb siv cov txheej txheem luam tawm yav dhau los [17]. Txhawm rau ntsuas qhov tseem ceeb, cais lub hom phiaj thiab kev tshawb nrhiav thiab suav cov qhab nia raws li qhov tsis tseeb nrhiav pom (FDRs) tau siv. Thaum kawg, peb lim cov ntaub ntawv SEQUESToutput los muab cov qhab nia kawg rau cov proteins. Yam tsawg kawg nkaus ntawm cov qhab-nees sib raug zoo (Xcorr) ntawm 1.5, 2.0, 2.25, thiab 2.5 raug xaiv rau ib leeg-, ob npaug-, triple-, thiab plaub-thooj-them ions, feem. Yav dhau los cov ntawv luam tawm tsis tau siv los lav qhov kev sib tw siab [20], thiab qhov tsis tseeb-zoo peptide piv yog tsawg dua 3 feem pua.

2.4. Kev txheeb cais.

Qhov normalized spectral abundance factor (NSAF) tau siv los xam cov txheeb ze ntawm polypeptides [21]. Kev hloov pauv thiab ntsuas qhov ntsuas peptide tau ua ua ntej kev txheeb xyuas txheeb cais. MetaboAnalyst 2. 05 [22] ib. *e piv ntawm ib tus neeg cov protein rau tag nrho cov concentration tau soj ntsuam siv cov tub ntxhais kawm ntawv t-test rau txhua pawg. PLS-DA thiab VIP tau siv los txheeb xyuas cov protein sib txawv [23]. Peb xaiv cov proteins nrog qhov tsis tseeb nrhiav pom tus nqi (FDR) tsawg dua lossis sib npaug li 10 feem pua ​​​​kom siv tau siv western blotting.

2.5. Western Immunoblotting thiab Quantification.

Cov tshuaj tiv thaiv tiv thaiv MGAM tau yuav los ntawm Proteintech Group, Inc., (Chicago, IL, USA) thiab tau siv los daws 100 ug ntawm cov protein los ntawm cov zis exosomes ntawm txhua qhov kev kawm. Tom qab sib cais, cov proteins raug xa mus rau cov ntawv nitrocellulose, thaiv, incubated nrog thawj cov tshuaj tiv thaiv ib hmos ua ntej ntxuav nrog Tris-buffered saline, incubated rau 1 h nrog HRP-secondary antibody conjugate, thiab pom los ntawm kev tsim raws li tau piav nyob rau hauv yav dhau los luam tawm los ntawm peb lub chaw kuaj mob [ 24, 25] ib. ImageJ software (NIH) tau siv los txheeb xyuas cov kab mob western immunoblot [24] thiab npaj (GraphPad Prism, San Diego, CA, USA).

3. Cov txiaj ntsig

3.1. Clinical Characteristics of Cirrhosis and Healthy Control Subjects.

Rau tus neeg mob hauv txhua pab pawg uas muaj cov ntaub ntawv kho mob tiav tau raug tshuaj xyuas thiab muab piv rau rau rau kev tswj hwm kev noj qab haus huv. Cov pej xeem thiab etiology ntawm kab mob siab tau sau tseg nyob rau hauv Table 1. Raws li kev cia siab raws li kev kawm tsim, Cov qhab nia ntawm Child Turcotte-Pugh thiab MELD sib txawv ntawm cov neeg mob.

3.2. Proteomic Analyzes Urine Exosomes los ntawm Cirrhotic Patients thiab Tswj Kev Noj Qab Haus Huv.

Nyob rau hauv tag nrho, thoob plaws tag nrho 4 pawg, 1572 cov proteins tshwj xeeb tau txheeb xyuas. Muaj 360 cov proteins uas muaj rau txhua pawg. Peb pom 83 qhov tshwj xeeb exosomal proteins rau pawg 0 (tswj), 250 rau pawg 1, 84 rau pawg 2, thiab 212 rau pawg 3 (Daim duab 1). Peb txuas ntxiv ua ntau yam sib txawv PLS-DA ntawm cov proteins, uas pom tau tias muaj kev sib cais meej ntawm pawg tswj kev noj qab haus huv thiab cov pab pawg sib txawv ntawm cov kab mob cirrhotic, raws li pom hauv daim duab 2. Kev them nyiaj thiab decompensated cirrhotic yam tsis muajmob raum(pab pawg 1 thiab 2) tau pom ntau qhov sib tshooj thaum cov ntsiab lus cirrhotic nrog AKI (pab pawg 3) pom meej meej sib cais los ntawm lwm yam mob cirrhotic thiab kev noj qab haus huv. Ib qho ANOVA cais ntawm cov proteins ntawm plaub pawg tau pom tias 126 cov proteins tau hloov pauv loj heev (p < 0.05),="" ntawm="" 13="" mus="" txog="" qhov="" kev="" tshawb="" pom="" cuav="" (fdr)="" txiav="" tawm=""><10% (table="" 2).="" maltase-glucoamylase="" (mgam)="" was="" the="" top="" discriminant="" protein="" with="" a="" vip="" score="" of="" 4.35="" for="" the="" entire="" study="">

3.3. Maltase-Glucoamylase Protein tau nce ntxiv hauv cov zis tso zis tso quav tso zis Exosomes.

Cov ntaub ntawv proteomic pom tias muaj ntau dua ntawm MGAM nyob rau hauv cov zis exosomes ntawm decompensated cirrhotic cov neeg mob, nrog thiab tsis.mob raum(pab 2 thiab 3). Qhov no yog ib qho kev ntxub ntxaug tshaj plaws ntawm tag nrho plaub pawg nrog tus qhab nia VIP ntawm 4.35 (Daim duab 3). Confirmatory western blotting ntawm cov exosomes pom pom muaj protein ntau nyob rau hauv cov neeg mob cirrhotic nrogmob raum(pab pawg 3) (Daim duab 4).

4. Kev sib tham

Peb tau ua qhov kev soj ntsuam proteomic ntawm cov ntsiab lus urinary exosome los ntawm cov neeg mob uas them nyiaj cirrhosis, decompensated cirrhosis, thiab decompensated cirrhosis nrog AKI thiab muab piv rau kev noj qab haus huv. Kev tsom xam proteomic ntawm urinary exosomes hauv cov neeg mob cirrhotic tau txheeb xyuas ntau yam tseem ceeb ntawm biomarkers ntawmmob raum, tshwj xeeb tshaj yog MGAM, ib qho enzyme bifunctional. Peb pom qhov siab tshaj plaws ntawm MGAM hauv cov zis exosomes ntawm cov neeg mob cirrhosis thiab AKI. Tsis tas li ntawd, MGAM tau nce ntxiv hauv cov neeg mob ntshav siab, tab sis tsis yog li ntawm cov neeg mob AKI. MGAM tsis nyob hauv pawg tswj hwm kev noj qab haus huv, qhia txog nws lub luag haujlwm muaj peev xwm ua biomarker ntawm kev mob hnyav.

Txoj kev tshawb no muaj ob peb lwm qhov kev tshawb pom tseem ceeb. Ua ntej, rau peb txoj kev paub, qhov no yog thawj daim ntawv tshaj tawm txog kev piav qhia ntawm cov zis exosome protein cov ntsiab lus nyob rau hauv cov yam ntxwv zoo cirrhotic. Qhov thib ob, qhov no yog thawj txoj kev tshawb fawb qhia txog kev nce tubular epithelial disaccharidase nyob rau hauvcirrhotic- raum raug mobparadigm. Cov ntaub ntawv tso zis exosome proteomic los ntawm 4 pawg sib txawv tau pom MGAM upregulation nyob rau hauv cirrhosis AKI pab pawg kom muaj zog thiab zoo ib yam. Maltase yog qhov loj disaccharidase nyob rau hauv lub raum txhuam ciam teb daim nyias nyias [26, 27], tab sis qhov tseeb ua haujlwm ntawm cov enzyme no tsis meej meej elucidated; Txawm li cas los xij, lub luag haujlwm ua tau rau cov kev cuam tshuam cuam tshuam, sucrase-isomaltase, thiab trehalase hauv cov piam thaj thauj tau raug xa mus [28]. Hauv kaum hom tsiaj, disaccharides ntsig txog MGAM tau pom nyob rau hauv lub raum txhuam ciam teb [27]. Farquhar thiab cov npoj yaig tau ua pov thawj tias maltase muaj nyob rau hauv microvilli ntawm lub proximal convoluted tubule, tej zaum ua haujlwm hauv qabzib reabsorption thiab thauj; thiab lub peev xwm nqus dej no txo ​​qis rau ntau qhov chaw ntawm lub nephron [29]. MGAM tau pom tias muaj nyob rau hauv exosomes thiab microparticles hauv tus qauv nas ntawm nonalcoholic steatohepatitis (NASH) [30], ib qho ua rau mob cirrhosis. Txij li cov neeg mob cirrhotic muaj ntau yam hauv qab uas ua rau txo qis hauv Scr, qhov kev tshawb pom ntawm AKI yog teeb meem. Tsis tas li ntawd, etiology ntawm kev raug mob feem ntau tsis paub, thiab kev sib txawv ntawm hepatorenal syndrome thiab lwm yam etiologies ntawm AKI yog qhov nyuaj. Tsis tas li ntawd, pathophysiology ntawm decompensated cirrhosis nrog AKI tsis paub meej vim tias nws tuaj yeem cuam tshuam cov kev hloov pauv ntawm hemodynamic lossis inflammatory mediators nyob rau hauv cov ntaub ntawv ntawm mob rau lub siab tsis ua hauj lwm [31]. Peb xav tias qhov kev hloov pauv ntawm cov qauv thiab kev ua haujlwm uas cirrhosis thiab portal hypertension coj los rau hauvlub raum ua haujlwmsib txawv ntawm qhov hnyav raws li qhov hnyav ntawm cirrhosis. Cov kev sib txawv ntawm cov nyiaj them rov qab thiab decompensated daim siab kab mob nrogmob raumtuaj yeem nkag siab los ntawm kev kawm cov khoom qis qis ntawm cov khoom sivraum, xws li zis. Muab cov nephron cell-xeev-xeev cov khoom thauj ntawm cov zis exosome, peb xav tias cov zis exosome tsom xam tuav cov ntaub ntawv tseem ceeb uas cuam tshuam rau qhov sib txawv ntawm AKI hauv cirrhosis. Daim ntawv tshaj tawm no yog thawj kauj ruam ntawm kev sim qhov kev xav no.

Txoj kev tshawb nrhiav no muaj ntau yam kev txwv. Ua ntej, tus naj npawb ntawm cov kev tshuaj ntsuam xyuas ib pab pawg tsawg. Cov qauv loj dua tuaj yeem ua kom muaj zog ntawm cov ntaub ntawv no ntxiv, thiab cov proteins ntxiv tuaj yeem mus txog qhov pib ntawm FDR uas tau txais.<10%. however,="" despite="" this="" small="" sample,="" these="" data="" demonstrating="" mgam="" as="" a="" unique="" exosomal="" protein="" in="" cirrhotic="" patients="" with="" aki="" is="" robust.="" second,="" we="" used="" 1d="" gel="" electrophoresis="" to="" resolve="" the="" exosome="" proteins="" prior="" to="" lc/ms-ms="" analysis="" that="" resulted="" in="" the="" identification="" of="" 1572="" proteins="" overall.="" if="" we="" had="" conducted="" direct="" exosome="" protein="" trypsinization="" instead="" of="" following="" this="" method,="" perhaps="" the="" number="" of="" identified="" proteins="" might="" have="" increased.="" in="" our="" experience,="" exosomes="" are="" packaged="" with="" nonfull="" length="" peptides="" from="" proteolytic="" action="" as="" well="" as="" endogenous="" peptides="" and="" may="" have="" con-founded="" the="" analysis.="" by="" following="" the="" gel="" electrophoresis="" method,="" we="" ensured="" that="" we="" only="" compared="" full-length="" protein="" differences="" between="">

Hauv cov ntsiab lus, peb tau pom cov zis exosome protein sib txawv hauv kev tswj kev noj qab haus huv thiab them nyiaj thiab decompensated cirrhotic cov ntsiab lus nrog thiab tsis muaj AKI los ntawm txoj kev proteomic. Kev ua haujlwm los ntawm pawg Knepper qhia tau hais tias ntau lub raum cov protein tseem ceeb (piv txwv li, aquaporins, polycystins, thiab podocin) raug tso rau hauv cov zis exosome [32, 33]. Peb daim ntawv tshaj tawm tam sim no ntxiv MGAM rau cov pab pawg no ntawm cov kab mob hauv lub raum tseem ceeb uas tau txheeb xyuas hauv cov zis exosomes. Peb qhov kev tshawb pom qhia tias MGAM tuaj yeem sib txawv ntawm cov tubular raug mob los ntawm lwm hom AKI hauv cov neeg mob cirrhotic. Txawm li cas los xij, qhov tseem ceeb ntawm kev kho mob ntawm MGAM upregulation nyob rau hauv cov kev kawm cirrhosis nrog AKI yuav tsum tau tsim nyob rau hauv cov kev tshawb fawb yav tom ntej.

Cov ntaub ntawv muaj

Cov ntaub ntawv kho mob thiab cov ntaub ntawv pov thawj siv los txhawb qhov kev tshawb pom ntawm txoj kev tshawb no yog txwv los ntawm UCSD lub koom haum tshuaj xyuas pawg thawj coj saib xyuas txhawm rau tiv thaiv tus neeg mob ntiag tug.*e cov ntaub ntawv muaj los ntawm Dr. Linda Awdishu (lawdishu@ucsd.edu) rau cov kws tshawb fawb uas ua tau raws li cov txheej txheem rau kev nkag mus rau cov ntaub ntawv tsis pub lwm tus paub.

Kev tsis sib haum xeeb

Cov kws sau ntawv tshaj tawm tias lawv tsis muaj kev tsis sib haum xeeb.

Kev lees paub

Kev tshawb fawb txhawb nqa los ntawm NIH NIDDK UAB / UCSD O'Brien Center Grant DK0793337.

Tus sau

Linda Awdishu,^1,2 Shirley Tsunoda,¹ Michelle Pearlman,³ Chanthel Kokoy-Mondragon,²Majid Ghassemian,⁴Robert K. Naviaux,⁵Heather M. Patton,³Ravindra L. Mehta,²Bhavya Vijay,⁶thiab Satish P.Ramachandra Rau ^2,6,7

1UC San Diego Skaggs Tsev Kawm Ntawv ntawm Pharmacy thiab Pharmaceutical Sciences, San Diego, USA

2 Biomarkers Laboratory, O'Brien Center rauMob raum mobKev tshawb fawb, Nephrology-Hypertension, UC San Diego, Department of Medicine, San Diego, USA

3UC San Diego, Department of Medicine, Division of Gastroenterology, San Diego, USA

4UC San Diego, Department of Chemistry & Biochemistry, Biomolecular & Proteomics Spectrometry Facility, San Diego, USA

5UC San Diego, Departments of Medicine, Pediatrics thiab Pathology, San Diego, USA

6I-AIM Biomarkers Laboratory, Re University of Trans-Disciplinary Health Sciences thiab Technology (TDU), Bangalore, Is Nrias teb

7UC San Diego, Department of Medicine, Division of Infectious Diseases, San Diego, USA

Kev sau ntawv yuav tsum tau hais rau Satish P. Ramachandra Rao; Tau txais 30 Lub Kaum Hli 2018; Hloov kho 31 Kaum Ob Hlis 2018; Tau txais 19 Lub Ob Hlis 2019; Luam tawm 16 Plaub Hlis 2019 Academic Editor: Antonio Artigas

Copyright © 2019 Linda Awdishu et al. Qhov no yog ib tsab xov xwm qhib-access faib nyob rau hauv lub,uas tso cai rau kev siv tsis txwv, kev faib tawm, thiab luam tawm hauv ib qho nruab nrab uas muab cov haujlwm qub raug suav hais tias yog.

Cov ntaub ntawv

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