Kev noj ntsev ntau kom txo tau lub raum cov ntaub so ntswg ntawm Inflammatory Cytokines hauv nas

edmund.chen@wecistanche.com

Abstract

Kev noj ntsev ntau (HS) feem ntau suav hais tias yog qhov ua rau ua rau muaj qhov ua rau mobmob raum. Txawm li cas los xij, cov kev hloov hauv lublub raumtheem ntawm inflammatory cytokines thaum lub sij hawm noj HS tseem tsis tau meej meej. Peb xav tias HS ncelub raumtheem ntawm qog necrosis factor-alpha (TNF-) thiab interleukin -6 (IL-6) tab sis de- creases interleukin -10 (IL-10; anti-inflammatory cytokine) thiab cov lus teb no exacerbate nyob rau hauv TSIS tsis muaj tej yam kev mob. Ob hom tsiaj qus (WT) thiab endothelial NO synthase knockout (eNOSKO) nas (~ 8 lub lis piam, n {{10}} hauv txhua pab pawg) tau muab ntsev ib txwm (NS; 0.3 feem pua ​​NaCl) thiab HS (4 feem pua ​​NaCl) muaj cov khoom noj rau 2 lub lis piam. Systolic ntshav siab (SBP) tau txiav txim siab los ntawm tus Tsov tus tw-cuff plethysmography thiab cov zis sau tau ua los ntawm cov kab mob metabolic. Basal SBP tau siab dua hauv eNOSKO dua WT nas (131 ± 7 vs 117 ± 3 mmHg; p < .05).="" hs="" tau="" txais="" 2="" lub="" lis="" piam="" nce="" sbp="" hauv="" enosko="" (161="" ±="" 5="" mmhg)="" tab="" sis="" tsis="" nyob="" hauv="" wt="" nas.="" hauv="" ns="" pawg,="" cov="" qib="" cytokine="">lub raumcov ntaub so ntswg (ntsuas siv ELISA cov khoom siv thiab qhia hauv pg/mg protein) tau ntau dua hauv eNOSKO dua WT nas (TNF- , 624 ± 67 vs. 325 ± 73; IL-6, 619 ± 106 vs. 166 ± 61 ; IL-10, 6,087 ± 567 vs. 3,929 ± 378). Qhov zoo siab, cov qib cytokine hauv HS pawg tau tsawg dua ob qho tib si hauv WT (TNF-, 114 ± 17; IL-6, 81 ± 14; IL-10, 865 ± 130) thiab eNOSKO (TNF- , 115 ± 18; IL-6, 56 ± 7; IL-10, 882 ± 141) nas. Cov kev tshawb pom no qhia tias HS induces downregulation ntawm cytokines nyob rau hauv lubraum.Xws li HS-vim txo qis hauv cytokines, tshwj xeeb tshaj yog TNF- (tus neeg sawv cev natriuretic), yuav pab kom muaj ntsev ntau dua, thiab yog li, ua rau ntsev-rhiab siab kub ntxhov nyob rau hauv TSIS muaj qhov tsis txaus.

Ntsiab lus:endothelial nitric oxide synthase, siab ntsev kom tsawg, IL-10, IL-6, nitric oxide, TNF-, raum, raum

CISTANCHE yuav txhim kho lub raum / raum tsis ua haujlwm

Taw qhia

Nws feem ntau pom tau tias kev noj zaub mov muaj ntsev ntau (HS) cuam tshuam nrog ntau yam kab mob plawv thiab cov kab mob metabolic xws li kub siab thiab ua rau mob hnyav. Cov ntaub ntawv pov thawj tsis muaj tsawg los qhia tias kev noj zaub mov HS cuam tshuam nrog ntau dua biomarkers ntawm o. Ntau cov kev tshawb fawb tsis ntev los no kuj tau pab txhawb rau qhov tseeb tias kub siab yog ib qho mob inflammatory kab mob uas muaj qhov tsis sib xws ntawm pro-inflammatory thiab anti-inflammatory cytokines. Qhov kev tsis txaus ntseeg no yog tshwm sim los ntawm dysregulation ntawm renin-angiotensin system (RAS) thiab oxidative kev nyuaj siab, uas cuam tshuam nrog kev noj ntsev ntau ntxiv thiab kev laus (Das, 2006; Guzik li al., 2007; Mattson, 2014; Rodriguez-Iturbe thiab al. , 2004). Tsis yog tsuas yog ib qho inducer ntawm oxidative kev nyuaj siab, tab sis angiotensin II (AngII) kuj tseem paub tias yuav ua rau muaj kev tsim cov kab mob cytokines xws li qog necrosis factor-alpha (TNF-) thiab interleukin (IL-6) (Funakoshi li al. ., 1999; Han et al., 1999; Ruiz-Ortega et al., 2002; Sanz-Rosa et al., 2005). Hauv ex-periments nrog cov qauv nas, kev noj zaub mov ntev ntawm HS tsis hloov cov ntshav siab (BP) (Kopkanet al., 2010; Lara et al., 2012). Txawm li cas los xij, HS kev noj zaub mov pub rau nas-kho tsis tu ncua nrog AngII lossis NO synthase (NOS) inhibitor, nitro-L-arginine-methyl ester (L-NAME) lossis hauv endothelial NOS knockout (eNOSKO) nas exaggerated cov lus teb hypertensive, thiablub rauminflammatory ntaub so ntswg raug mob (Francois li al., 2008; Kopkan et al., 2010; Kopkan & Majid, 2005; Nakamura et al., 2011; Singh et al., 2013, 2017). HS noj nyob rau hauv eNOSKO nas tau pom tias yuav txhawb kev kub siab thiabmob raumtxuam nrog nce oxidative kev nyuaj siab thiab zus tau tej cov AngII (Nakamura li al., 2011). Cov txiaj ntsig no qhia tau hais tias HS kev noj haus cuam tshuam rau kev tsim cov cytokines inflammatory nyob rau hauv cov xwm txheej ntawm oxidative kev nyuaj siab los yog nyob rau hauv kev txhim kho RAS kev ua. Peb tau tshaj tawm yav dhau los tias qhov mob hnyav ntawm NOS los ntawm kev ua haujlwm ntawm L-NAME hauv cov nas ua rau cov ntshav nce ntxiv thiablub raumtheem ntawm TNF- (Shahid et al., 2010; Singh li al., 2014), nrog nce infiltration ntawm macrophages hauv lubraum(Islam et al., 2011). Xws li cov kab mob NOS inhibition los ntawm mob L-NAME infusion hauv nas cov qib ntawm cov tshuaj tiv thaiv cytokine, interleukin -10 (IL-10) hauv cov ntshav thiablub raumcov ntaub so ntswg (Singh et al., 2014). Txawm li cas los xij, qhov kev hloov pauv ntawm cov kab mob cytokine nyob rau hauv cov lus teb rau kev noj tshuaj HS ntev nyob rau hauv ib txwm muaj thiab cov xwm txheej ntawm NO tsis txaus tseem tsis tau txiav txim siab meej.

Peb xav tias HS tau txais txiaj ntsig zoo dualub raumtheem ntawm pro-inflammatory thiab txo cov tshuaj tiv thaiv cytokines, thiab cov lus teb no exacerbate nyob rau hauv cov mob tsis muaj peev xwm. Txhawm rau tshuaj xyuas qhov kev xav no, peb tsom los ntsuas qhov kev hloov pauv ntawm cov xwm txheej ntawm cov kab mob cytokines, tshwj xeeb hauvraumThaum noj cov zaub mov HS, thiab txiav txim siab seb cov cytokines no cuam tshuam li cas los ntawm qhov tsis muaj peev xwm. Hauv qhov kev tshawb fawb no, peb ntsuas qhovlub raumas well as plasma level of pro(IL{{0}}} and TNF-) and anti- (IL-10) inflammatory cytokines in wild-type (WT) and eNOSKO nas uas tau noj li qub- ntsev (NS; 0.3 feem pua ​​NaCl) thiab HS (4 feem pua) uas muaj cov khoom noj rau 2 lub lis piam.

CISTANCHE yuav txhim kho lub raum / raum kab mob

2. Cov khoom siv thiab cov txheej txheem

Cov kev sim no tau ua raws li cov lus qhia thiab kev coj ua uas tsim los ntawm Tulane University Animal Care and Use Committee. Txiv neej eNOSKO nas (B6.129P2-Nos3tm1Unc/J; stock no: 002684) nas thiab lawv cov kab mob keeb kwm yav dhau los ntawm WT (C57BL/6J; stock no: 002684) nas (Jackson Laboratory, Bar Harbor, ME) suav tias yog kev tswj hwm tau siv hauv txoj kev tshawb no. Cov nas no tau nyob hauv chav ntsuas kub- thiab lub teeb-tswj thiab tso cai pub dawb rau kev noj zaub mov zoo (Ralston-Purina, St. Louis, MO) thiab dej kais. Cov nas muaj hnub nyoog 8-9 lub lis piam; ∼20–22 g lub cev hnyav) tau muab faib ua ntau pawg nyob ntawm kev noj zaub mov NS (ib qho khoom noj uas muaj 0.3 feem pua ​​NaCl) thiab HS noj (HS, 4 feem pua ​​NaCl; Harlan-Teklad, Madison, WI), feem. , muaj 6 tus tsiaj hauv txhua pab pawg. Cov pab pawg muaj raws li nram no: 1. WT-NS: Cov nas qus tau noj NS, 2. WT-HS: Cov nas qus tau noj HS noj 3. eNOSKO-NS: eNOSKO nas noj NS noj, 4. eNOSKO-HS: eNOSKO nas tau noj HS noj

2.1 Kev saib xyuas BP hauv cov nas nco qab

Lub systolic BP (SBP) tau ntsuas hauv cov nas sim siv cov txheej txheem noninvasive tail-cuff plethysmography (Visitech Systems, Apex, NC). Cov nas raug saib xyuas rau 7–10 hnub ntawm kev sim zaub mov, thiab basal BP raug kaw ua ntej hloov mus rau cov zaub mov muaj ntsev (tsuas yog rau WT-HS thiab eNOSKO-HS nas) rau qhov seem ntawm txoj kev kawm. Hauv cov nas no, BP tau ntsuas tib lub sijhawm ntawm hnub (11 teev sawv ntxov txog 1 teev tsaus ntuj) kom tsis txhob muaj kev cuam tshuam ntawm lub voj voog circadian, thiab tus nqi ntawm SBP tau txais los ntawm kev kwv yees qhov nruab nrab ntawm kev nyeem ntawv ntawm 10 kev ntsuas rau ib leeg. kev sim. Cov nas tau raug cob qhia rau tail-cuff BP ntsuas 7-10 hnub ua ntej pib qhov kev sim. Tom qab 2-3 hnub ntawm acclimatization rau tus Tsov tus tw-cuff ntaus ntawv, lub basal SBP tau ntsuas nyob rau hauv tag nrho cov nas nyob rau hauv standard laboratory noj zaub mov ua ntej pib ntawm HS noj thiab xam raws li hnub 0 tus nqi. Tom qab ntawd, cov tsiaj ntawm HS pawg tau hloov mus rau HS kev noj haus rau qhov seem ntawm txoj kev tshawb no (tom qab ntawd SBP tau ntsuas rau hnub 3, 6, 9, thiab 12 ntawm NS / HS noj) ntawm 12: 12hr lub voj voog tsaus ntuj thiab tau txais zaub mov thiab dej ad libitum thoob plaws qhov kev kawm.

2.2 Kev kho tsiaj

Tom qab sau tseg ntawm so SBP, nas tau noj nrog NS ({{0}}}.3 feem pua ​​NaCl) lossis HS (4 feem pua ​​NaCl) noj rau 2 lub lis piam los ntsuas cov lus teb rau HS ib leeg hauv WT thiab eNOSKO nas. Ob qho khoom noj no muaj kwv yees li qhov sib npaug ntawm lwm cov as-ham (19 feem pua ​​​​cov protein, 5 feem pua ​​​​ rog, 3 feem pua ​​​​ crude fiber, 0.86 feem pua ​​​​Ca, 0.64 feem pua ​​P, { {13}}.72 feem pua ​​K, thiab 0.15 feem pua ​​Mg). Thaum lub sijhawm sim, cov nas tau tso cai haus dej dawb los ntawm cov fwj dej uas txuas nrog lub tawb. Kev noj zaub mov thiab dej, nrog rau lub cev hnyav, ntawm txhua tus nas raug ntsuas thaum lub sijhawm sim.

2.3 Kev sau thiab tshuaj xyuas cov zis thiab cov ntshav kuaj

Nees nkaum plaub teev cov zis tau sau los ntawm cov nas uas tsis nco qab siv cov kab mob metabolic nyob rau hnub basal (Hnub 0) thiab hnub kawg (Hnub 13) txhawm rau ntsuas qhov excretory tsis muaj lossis tsis muaj HS noj hauv cov nas no. Tag nrho cov pab pawg nas tau muab tso rau hauv cov kab mob metabolic thiab tso zis, thiab cov kev ntsuas tsim nyog tau sau rau tib lub sijhawm. Tsiaj txhu tau nyob ib leeg nyob rau hauv cov kab mob metabolic, thiab cov zis tau khaws cia rau 24 teev rau hauv cov hlab tsis muaj menyuam. Cov zis ntim tau txiav txim siab los ntawm txhua qhov tso zis, thiab cov qauv tau centrifuged (3, 000 rpm / 5 min; 4 degree) thiab khaws cia rau kev txiav txim siab tso zis ntawm sodium thiab potassium. Kev tso zis ntawm sodium thiab potassium raug soj ntsuam los ntawm nplaim hluav taws xob (Singh li al., 2013, 2017).

Thaum kawg ntawm lub sijhawm sim, tag nrho cov tsiaj tau euthanized, thiab cov ntshav tau sau. Plasma tau muab cais tawm tam sim ntawm cov ntshav thiab khaws cia ntawm −80ºC rau kev kwv yees ntawm pro- (IL-6 & TNF- ) thiab anti- (IL-10) inflammatory cytokines raws li tau ua ua ntej (Singh li al., 2014, 2017). Covlub raumkuj raug tshem tawm. Ibraumtau kho nyob rau hauv 10 feem pua ​​​​ntawm cov tshuaj formalin thiab paraffin-embedded rau kev ntsuam xyuas ntawmmob raum. Lwm tusraumtau ua tiav los ntsuas cov qib ntawm inflammatory cytokines. A 100 mgraummuaj ob qho tib si cortical thiab medullary cheeb tsam tau homogenized nyob rau hauv sterile phosphate-buffered saline uas muaj protease inhibitor ntawm 4 degree. Covraumhomogenates tau centrifuged ntawm 9, 000 g rau 10 min ntawm 4 degree . Cov supernatants raug xa mus rau kev ntxuav cov hlab micro-centrifuge thiab khaws cia ntawm −80 degree kom txog rau thaum kuaj xyuas.

2.4 Enzyme-linked immunosorbent assay rau IL-10, IL-6, thiab TNF- theem

Qib IL-10, IL-6, thiab TNF- hauv ntshav thiab supernatants los ntawmraumCov ntaub so ntswg homogenates tau ntsuas siv ELISA Ready-SET-go cov khoom siv (eBioscience, Inc., San Diego, CA). Cov qib kuaj pom ntawm cov khoom siv (tus qauv nkhaus ntau) yog raws li hauv qab no: IL-10 cov khoom siv (Catalog no. 88-7105-88), 32–4000 pg/ml; IL-6 Cov khoom siv (Catalog no. 88-7064-22), 4–500 pg/ml; TNF- Cov Khoom Siv (Catalog no. 88-7324-86), 8–1000 pg/ml. Cov theem ntawm cytokines hauvlub raumCov ntaub so ntswg tau normalized los ntawm cov protein ntau (tseem ntsuas los ntawm Bio-Rad detergent tau tshaj cov protein soj ntsuam txoj kev) (Cat. No. # 500-0112; Bio-Rad, Hercules, CA)

2.5 Kev ntsuam xyuas ntawm lub raum raug mob

Formalin-taw paraffin-embeddedraumcov seem tau siv rau kev tshuaj xyuas glomerulosclerosis siv periodic acid-Schiff staining (Lara li al., 2012; Luna, 1992; Singh li al., 2013, 2017) thiab tubulointerstitial fibrosis siv Gomori's triom50 staining, 9. 2013, 2017).

CISTANCHE yuav txhim kho lub raum / raum mob

2.6 Kev kwv yees ntawm glomerulosclerosis

Tus nqi ntawm glomerulosclerosis tau soj ntsuam ntau npaum li cas los ntawm kev txheeb xyuas cov duab tsis siv neeg ntawm txhua lub glomerulus siv periodic-acid-Schiff stainedlub raumseem (Mass Histology Service, Worcester, MA) (Lara li al., 2012; Luna, 1992; Singh li al., 2013, 2017). Nees nkaum daim duab los ntawm txhuaraumswb nrog tsawg kawg ib lub glomerulus ib daim teb tau yees duab siv Nikon Eclipse 50i microscope nruab nrog Nikon DS Camera Head (DS Fi1) thiab DS lub koob yees duab tswj chav tsev (DSU2). Ib qho xim liab doog hauv glomerulus tau lees paub tias yog sclerosis. Qhov feem pua ​​​​ntawm cov cheeb tsam uas tau them los ntawm cov kab mob sclerosis hauv glomeruli hauv txhua daim teb tau txheeb xyuas siv Nikon NIS-Elements software (version 2.34). Qhov feem pua ​​​​ntawm cov ntaub ntawv tau txais rau txhua ntawm 20 cov duab tau nruab nrab kom tau txais qhov feem pua ​​​​ntawm cov kab mob sclerosis rau tag nrho cov swb.

2.7 Kev kwv yees ntawm tubulointerstitial fibrosis

Qhov cuam tshuam ntawm interstitial collagen-zoo cheeb tsam (fibrosis) tau soj ntsuam ntau npaum li cas los ntawm cov duab tsis siv neeg tsom xam ntawmlub raumseem nyob los ntawm interstitial cov ntaub so ntswg staining zoo rau collagen nyob rau hauv Gomori lub trichrome stained seem (Gomori, 1950; Singh li al., 2013, 2017). Formalin-taw paraffin-embeddedlub raumcov seem tau stained nrog plasmin stain (chromotrope 2R) thiab cov ntaub so ntswg sib txuas (aniline xiav) ua ke hauv cov tshuaj phosphotungstic acid uas glacial acetic acid tau ntxiv. Qhov no stained lub collagen xiav, uas qhia tias fibrosis. Slides tau yees duab raws li tau piav saum toj no. Qhov feem pua ​​​​ntawm cov khoom siv collagen hauv txhua daim teb tau txheeb xyuas siv Nikon NIS-Elements software (version 2.34). Qhov feem pua ​​​​ntawm cov ntaub ntawv tau txais rau txhua ntawm 20 cov duab tau nruab nrab kom tau txais qhov feem pua ​​​​ntawm cov kab mob fibrosis rau tag nrho cov swb.

2.8 Kev txheeb cais

Tag nrho cov txiaj ntsig tau qhia raws li txhais tau tias ± SE. Kev txheeb xyuas kev txheeb xyuas tau ua tiav siv Sigma stat software (Systat Software, hnub nyob rau hauv 2 lub lis piam ntawm kev kawm yam tsis tau sau cov quav thiab lwm yam kev ntsuas ntawm insensible poob ntsev thiab dej. Nws tsis tuaj yeem ua los ntawm cov khoom sib sau los ntawm cov kab mob metabolic.

3.3 Inflammatory cytokine theem

3.31 TSI-Daim duab 2 qhia txog lub plasma thiablub raumcov ntaub so ntswg ntawm TNF- (Daim duab 2a thiab b) thaum kawg ntawm lub sijhawm sim. Qhov TNF- concentration hauv cov ntshav tsis tau kuaj pom hauv NS cov pab pawg, ob leeg WT thiab eNOSKO nas. Txawm li cas los xij, hauv HS cov pab pawg ntawm cov nas, cov plasma TNF- qib tau kuaj pom muaj qhov hloov pauv siab hauv WT, 202 ± 146 pg / ml thiab eNOSKO, 175 ± 68 pg / ml) nas. Hauv cov pab pawg NS noj, TNF- qib hauv lub raum cov ntaub so ntswg yog siab dua hauv eNOSKO dua WT nas (624 ± 67 vs. 325 ± 73, pg/mg protein; p < .05).="" txawm="" li="" cas="" los="" xij,="" hauv="" pawg="" hs="" kev="" noj="">lub raumcov ntaub so ntswg TNF- qib tau qis dua hauv WT (114 ± 17 pg / mg protein) thiab eNOSKO (115 ± 18 pg / mg protein) nas piv rau NS noj cov pab pawg.

3.32 IB -6

Daim duab 3 qhia txog lub plasma thiablub raumcov ntaub so ntswg ntawm IL-6 (Daim duab 3a thiab b). Hauv pawg NS noj, IL-6 concentration hauv cov ntshav (180 ± 44 vs. 13 ± 11 pg/ml; p < .05)="" thiab="" hauv="">raum(619 ± 106 vs. 166 ± 61 pg / mg protein; p < .05)="" hauv="" enosko="" yog="" siab="" dua="" wt="" nas.="" txawm="" li="" cas="" los="" xij,="" plasma="" il-6="" qib="" hauv="" hs="" tau="" txais="" cov="" pab="" pawg="" tsis="" raug="" kuaj="" pom="" hauv="" wt="" thiab="" enosko="">Lub raumcov ntaub so ntswg IL-6 qib kuj tseem qis dua hauv HS noj cov pab pawg hauv ob qho tib si eNOSKO (56 ± 7 pg / mg protein) thiab WT (81 ± 14 pg / mg protein) nas piv rau NS noj cov pab pawg.

3.33 IB -10

Daim duab 4 qhia txog lub plasma thiablub raumcov ntaub so ntswg ntawm IL-10 (Daim duab 4a thiab b). Hauv NS noj cov pab pawg, plasma qib ntawm IL-10 hauv eNOSKO qis dua WT nas (510 ± 125 vs. 701 ± 54 pg / ml; p < .05).="" txawm="" li="" cas="" los="" xij,="">lub raumcov ntaub so ntswg IL-10 qib siab dua hauv eNOSKO dua WT nas (6,087 ± 567 vs. 3,929 ± 378, pg/mg protein; p < .05).="" nyob="" rau="" hauv="" hs="" noj="" pab="" pawg,="" plasma="">lub raumCov ntaub so ntswg ntawm IL-10 tau qis dua hauv eNOSKO (327 ± 67 pg / ml; 882 ± 141 pg / mg protein) thiab WT (344 ± 52 pg / ml; 865 ± 130 pg / mg protein) nas.

3.4 Kev raug mob rau lub raum tsis ua haujlwm

3.4.1|Glomerulosclerosis

Cov duab sawv cev ntawm periodic-acid-Schiff stainedraumcov seem uas muab qhov kev txiav txim siab ntawm glomerular sclerosis tau muab rau hauv daim duab 5a thiab feem pua ​​​​ntawm cov cheeb tsam sclerotic tau piav qhia hauv daim duab 5b. Txawm hais tias peb tau pom muaj kev cog lus thiab thrombosed glomeruli hauv NS-fed eNOSKO tab sis tsis yog WT nas, tsis muaj qhov sib txawv ntawm qhov feem pua ​​​​ntawm cov kab mob sclerotic ntawm WT-NS thiab eNOSKO-NS nas (11.4 ± 1 feem pua ​​thiab 11.2 ± 1 feem pua). Txawm li cas los xij, qhov feem pua ​​​​ntawm thaj chaw sclerotic qis dua hauv WT-HS piv rau WT-NS nas (7.7 ± 1 feem pua ​​​​vs. 11.4 ± 1 feem pua; p < .05).="" ntawm="" qhov="" tsis="" sib="" xws,="">

3.4.2 Tubulointerstitial fibrosis

Cov duab sawv cev ntawm Gomori's trichrome-stained seem muab qhov twglub rauminterstitial fibrosis tau muab rau hauv daim duab 6a, thiab feem pua ​​​​ntawm cov chaw fibrotic tau qhia hauv daim duab 6b. Zuag qhia tag nrho, cov txiaj ntsig fibrotic hauv lub raum interstitium yog qhov qis dua hauv cov qauv nas no. Hauv WT nas, thaj chaw fibrotic nyob hauvlub rauminterstitium tau zoo sib xws hauv ob pawg NS thiab HS noj. Txawm li cas los xij, raws li qhov xav tau, thaj chaw fibrotic tau nce siab dua hauv eNOSKO-NS piv rau WT-NS nas (2.2 ± 0.2 feem pua ​​​​vs. 1.5 ± 0.2 feem pua; p < .05)="" .="" txawm="" li="" cas="" los="" xij,="" thaj="" chaw="" fibrotic="" hauv="">lub rauminterstitium qis dua hauv eNOSKO- HS dua eNOSKO-NS nas (1.8 ± 0.1 feem pua ​​vs. 2.2 ± 0.2 feem pua ​​; p < .05).

4 Kev sib tham

Txawm hais tias tsis muaj qhov tsis muaj peev xwm ua rau muaj kev txhim kho hauv cov cytokines, cov txiaj ntsig ntawm txoj kev tshawb no qhia tau hais tias HS tau txais hauv cov nas, tsawg kawg yog 2 lub lis piam, feem ntau txwv cov qib ntawm ob qho tib si pro- thiab anti-inflammatory cytokines, tshwj xeeb tshaj yog nyob rau hauv cov nas.raum. Cov qib plasma basal ntawm TNF- feem ntau tsis tuaj yeem kuaj pom hauv WT thiab eNOSKO nas uas tau noj NS cov zaub mov. Txawm li cas los xij, ib qho tseem ceeb ntawm cov cytokine no tau pom nyob rau hauv ob hom kev noj HS noj rau 2 lub lis piam. Yog li ntawd, nws qhia tau hais tias HS tau txais kev txhawb nqa kev tsim tawm ntawm TNF-, raws li qhia hauv lwm cov kev tshawb fawb (Costa li al., 2012; Liu li al., 2012; Yilmaz li al., 2012; Zhu et al., 2014). Txawm li cas los, nws yog nthuav kom nco ntsoov tias lublub raumtheem ntawm TNF- yog qhov qis dua hauv HS cov pab pawg ntawm WT thiab eNOSKO nas piv rau hauv pawg NS noj. Xws li HS-vim txo qis kuj tau sau tseg hauv lwm cov cytokines xws li IL-6 thiab IL-10 hauvraum.Txawm tias covlub raumtheem ntawm IL-6 thiab IL-10 tau siab dua hauv eNOSKO-NS nas dua li cov nyob hauv WT-NS nas, lawv cov ntshav plasma kuj sib txawv. Piv nrog rau WT-NS nas, plasma qib ntawm IL-6 tau siab dua, IL-10 qis dua, tab sis TNF- qib tsis txawv hauv eNOSKO-NS nas. Txawm li cas los xij, qhov siab dualub raumtheem ntawm pro-inflammatory cytokines hauv eNOSKO-NS nas yog me ntsis xav tau. TSIS yog ib qho kev taw qhia molecule uas feem ntau plays ib qho kev tiv thaiv kab mob nyob rau hauv ib txwm physiological mob (Harrison, 1997;

Moncada et al., 1991). Tsis muaj qhov tsis txaus yog feem ntau paub tias ua lub luag haujlwm tseem ceeb hauv kev ua rau cov txheej txheem inflammatory (Liu & Huang, 2008). Txawm li cas los xij, qib ntawm kev paub txog kev tiv thaiv kab mob cytokine, IL -10, kuj tseem siab dua hauvlub raumcov ntaub so ntswg ntawm eNOSKO-NS nas piv rau hauv WT-NS nas. Qhov no tuaj yeem yog vim qhov tseeb IL-10 tsis yog ib txwm ua raws li tus neeg sawv cev tiv thaiv kab mob tab sis kuj tuaj yeem ua raws li cov cytokine pro-inflammatory hauv qee yam mob uas txuas rau siab RAS, raws li tau pom ua ntej los ntawm peb lub chaw kuaj mob (Singh thiab ib., 2017).

Txawm hais tias nws feem ntau pom tau tias HS kev noj zaub mov feem ntau cuam tshuam nrog kev cuam tshuam ntawm biomarkers ntau dua ntawm kev mob, feem ntau ntawm cov kev tshawb pom no tau pom nyob rau hauv cov kev tshawb fawb pej xeem uas HS noj cov zaub mov feem ntau yog coded rau cov khoom noj uas muaj roj ntau thiab cov khoom noj muaj protein ntau, uas tuaj yeem ua rau qee qhov kev tshwm sim no (Zhu li al., 2014). Ib qho kev tshawb fawb tsis ntev los no kuj tau qhia tias kev noj cov ntsev rov qab zoo dua, tab sis tsis yog ntuj hiav txwv ntsev, ua rau kub siab thiab ua rau txawv txav.raumpathology hauv Dahl ntsev-sensitive nas (Lee li al., 2017). Yog li, muaj kev sib raug zoo ntawm kev noj ntsev thiab kev tiv thaiv kab mob hauv tib neeg lub cev. Txoj kev tshawb no yog thawj zaug ntawm cov kev tshawb fawb uas siv cov qauv tsiaj los qhia cov lus teb rau kev tswj hwm ntau dua ntawm HS cov zaub mov ntawm qib ntawm qee qhov mob cytokines hauv cov ntshav thiab hauv cov ntshav.raum.Qhov pom kev txo qis hauv covlub raumKev tsim cov cytokines thaum lub sij hawm HS tau txais tuaj yeem cuam tshuam nrog kev tawm tsam ntawm RAS vim HS tau txais (Drenjančević-Perić li al., 2011).

Nws raug sau tseg tias qib siab ntawm plasma IL-10 tau kuaj pom hauv NS-fed WT nas, tab sis nws qib tsuas yog qis me ntsis hauv NS-fed eNOSKO nas. Ntawm qhov tsis sib xws, qhov kev txo qis ntawm IL-10 tau pom nyob rau hauv cov lus teb rau mob NOS inhibition hauv nas, raws li qhia hauv peb txoj kev tshawb fawb dhau los (Singh li al., 2014). Txawm li cas los xij, IL-10 qib hauv plasma tseem qis dua hauv WT thiab eNOSKO nas thaum noj HS hauv txoj kev tshawb no. Cov ntaub ntawv no qhia tau hais tias kev noj tshuaj HS ntev feem ntau txwv cov qib ntawm ob qho tib si los tiv thaiv cytokines, tshwj xeeb hauvraum. Hauv eNOSKO nas,lub raumcov ntaub so ntswg nitrotyrosine qib qis dua piv nrog rau hauv WT nas (Kopkan li al., 2010) qhia tias peroxynitrite ntau lawm tsawg nyob rau hauv cov nas uas tsis muaj eNOS thiab qhov no yuav pab txhim kho IL-10 qib hauv cov nas no (Cook et al., 2003; Numanami et al., 2003). plasma ntau dua IL-10 qib tuaj yeem tsim cov nyhuv inhibitory ntawm TNF-plasma hauv eNOSKO nas, raws li nws tau pom tias qhov mob hnyav ntawm IL-10 hauv cov nas txo TNF- qib hauv covraum(Singh li al., 2014). Hauv qee qhov xwm txheej, txawm li cas los xij, IL-10 tau pom tias ua raws li tus neeg sawv cev tiv thaiv kab mob kom ntxiasmob raum,tshwj xeeb tshaj yog thaum nce angiotensin II hauv nas (Singh li al., 2017). Yog li, nws muaj peev xwm ua tau tias IL-10 ua raws li kev tiv thaiv cytokine hauv eNOS gene knockout hom.

Raws li ib txwm muaj, HS kev noj haus tsis hloov pauv BP hauv WT tab sis ua rau muaj kev nce ntxiv hauv BP hauv eNOSKO nas (Kopkan li al., 2010) uas lub hauv paus BP siab dua piv rau WT nas. Raws li HS kev noj haus ua rau txo qis hauv cytokine qib hauv ob hom nas, cov lus teb hauv eNOSKO nas yuav tsis qhia txog kev sib raug zoo ntawm BP hloov nrog cov kev hloov hauv cov cytokines inflammatory. Tseem tsis tau muaj pov thawj muaj nyob rau tam sim no, qhov kev hloov pauv hauv cov hlab ntsha ncaj qha cuam tshuam rau kev tsim cov TNF- tab sis ntawm qhov tsis sib xws, peb thiab lwm tus tau tshaj tawm tias cov tshuaj intravenous infusion ntawm tib neeg recombinant TNF- txo BP hauv cov nas (Kramer li al., 1988. ; Shahid et al., 2008). Hauv kev tshawb fawb tam sim no, kev hloov pauv zoo sib xws hauv plasma thiablub raumtheem ntawm TNF- thiab lwm yam cytokines hauv WT thiab eNOSKO nas tau pom los ntawm BP nce tsuas yog hauv eNOSKO nas thiab tsis nyob hauv WT nas. Qhov kev soj ntsuam no yuav tsis yog qhov xav tsis thoob li ntau lwm cov kev tshawb fawb txog kev cuam tshuam hauv tib neeg (Luo li al., 2016) kuj tau qhia txog qhov tsis muaj kev sib raug zoo nrog inflammatory cytokines thiab kub siab thaum noj HS. Hauv kev tshawb nrhiav cov placebo-controlled crossover nyob rau hauv cov kev kawm tsis tau kho (ua ntej) hypertensive cov ntsiab lus, nws kuj tau sau tseg tias lub sijhawm 4-lub lim tiam ntawm kev noj ntsev tsis cuam tshuam rau cov cim inflammatory xws li plasma qib ntawm TNF- , IL{{6} } , IL-6, IL-8, or MCP-1) (Gijsbers et al., 2015). Nws kuj tseem tau sau tseg tias BP hauv tib neeg tseem tsis tau hloov pauv thaum noj ntsev tsawg uas ua rau muaj kev tsim cov tshuaj tiv thaiv kab mob xws li TNF- , IL-6, lossis pro-calcitonin (Mallamaci li al., 2013). Yog li, kev sib raug zoo ncaj qha ntawm kev hloov pauv hauv cytokines thiab BP hloov pauv tseem tsis tau muaj pov thawj sim.

Peb cov kev tshawb fawb yav dhau los hauv cov nas tsuag (Kopkan et al., 2010; Kopkan & Majid, 2005; Singh et al., 2017) tau pom tias endogenous TSIS MUAJ ntau lawm los ntawm eNOS txo ntsev rhiab heev rau cov lus teb rau kev noj zaub mov HS. Cov kev tshawb fawb hauv cov tsiaj nyob hauv ib txwm muaj qhia tias kev noj HS ntev ib leeg ua rau tsis muaj lossis hloov pauv tsawg hauv BP. Txawm li cas los xij, HS kev noj haus ua rau ntsev rhiab heev thiab kub siab thaum tsis muaj kev tsim tawm hauv eNOSKO nas (Kopkan et al., 2010) lossis nas (Kopkan & Majid, 2005) kho mob ntev nrog NOS inhibitor, L-NAME. Peb qhov kev soj ntsuam hauv txoj kev tshawb fawb tam sim no qhia tias HS kev noj haus ua rau muaj ntsev-sensitive ntshav siab tsuas yog hauv eNOSKO nas tab sis tsis yog WT nas (Daim duab 1). Qhov no feem ntau yuav yog vim kev txhim kho hauv tubular sodium reabsorption hauv lubraumuas tshwm sim los ntawm kev nce cov ntaub so ntswg superoxide thaum tsis muaj kev tsim khoom los ntawm eNOS kev ua haujlwm raug cuam tshuam (Kopkan li al., 2010). Kev noj haus HS ntev feem ntau ua rau tsis muaj kev tsim khoom los ntawm kev tswj hwm eNOS kev ua (Mattson & Higgins, 1996). Xws li kev nce qib NO txo qis cov ntaub so ntswg superoxide ua haujlwm kom tswj tau qhov xwm txheej ntawm "oxidative tshuav nyiaj li cas" uas txhawb nqa cov sodium homeostasis, thiab yog li txo qis qhov nce hauv BP. Peb kuj tau ua pov thawj tias cov lus teb mob siab rau cov kab mob HS ntev npaum li cas hauv Il-10 cov nas knockout raug txo vim kev txhim kho hauv eNOS qhia hauv cov nas no (Singh li al., 2017). Kev nce hauv plasma TNF- theem thaum lub sijhawm noj HS ntev kuj tseem ua rau txo qis qhov kev txhim kho hauv tubular sodium reabsorption, vim qhov nce ntawm sodium load hauv WT nas txawm tias txo qis hauv cov ntshav.lub raumcov ntaub so ntswg theem ntawm TNF-. Txawm li cas los xij, nyob rau hauv qhov xwm txheej ntawm oxidative tsis txaus vim tsis muaj qhov tsis txaus, xws li kev tiv thaiv ntawm plasma TNF- theem yog cuam tshuam, ua rau txhim kho sodium tuav, thiab yog li, ua rau ntsev rhiab heev thiab kub siab hauv eNOSKO nas. Peb tau pom yav dhau los tias qhov nce hauv plasma TNF- theem ua rau cov lus teb natriuretic los ntawm kev ua kom cov tubular TNF- receptor hom 1 (TNFR1) (Castillo li al., 2012). Peb cov ntaub ntawv ua ntej tsis ntev los no kuj qhia tau hais tias TNFR1 cov protein qhia tau txhim kho thaum lub sijhawm noj HS ntev hauv WT nas tab sis txo qis hauv TSIS muaj qhov tsis txaus xws li hauv eNOSKO nas (Majid li al., 2018) lossis hauv cov nas uas tau kho nrog L-NAME (Majid. et al., 2017). Xws li kev txo qis hauv TNFR1 kev ua haujlwm kuj tseem ua rau muaj ntsev rhiab heev thiab kub siab thaum HS tau txais hauv eNOSKO nas. Yog li, nws pom tau tias eNOS muaj lub luag haujlwm tseem ceeb hauv kev txo qis qhov cuam tshuam ntawm HS ntev npaum li cas hauv WT nas.

Kev noj qab haus huv HS ntev, los ntawm "osmoreceptor muaj nuj nqi," ua rau lub cev tiv thaiv kab mob uas ua rau cov kab mob mononuclear phagocyte (MPS) cov hlwb hauv cov ntaub so ntswg myeloid hauv cov pob txha pob txha, spleen, thiab cov ntaub so ntswg ntawm daim tawv nqaij uas tom qab ntawd circulates thiab infiltrates ntau lub cev nrog rauraum(Machnik et al., 2009). Cov activated MPS hlwb tso tawm TNF- uas tshwm nyob rau hauv txoj kev raws li nws daim ntawv soluble (sTNF- ). sTNF- yog tsim los ntawm proteolytic cleavage los ntawm daim nyias nyias-tethered daim ntawv (mTNF- ) los ntawm TNF- -hloov enzyme (TACE), ib tug sheddase uas kuj tau txheeb xyuas raws li ib tug disintegrin thiab metalloprotease 17 (ADAM17) (Black li al. , 1997; Kriegler et al., 1988; Moss et al., 1997). Nrog rau kev tsim tawm ntawm TNF-, activated MPS hlwb kuj tso tawm NO feem ntau los ntawm eNOS ua kom (Connelly li al., 2003). Qhov kev soj ntsuam tseem ceeb ntawm qhov sib txawv ntawm TNF- cov lus teb hauv txoj kev tshawb fawb tam sim no qhia tias kev noj tshuaj HS ntev, ntawm ib sab, nce sTNF- hauv cov ntshav, tab sis txo qis mTNF- hauv cov ntshav.lub raumcov ntaub so ntswg, ntawm qhov tod tes, feem ntau yuav yog los ntawm kev ua kom TACE (Black li al., 1997; Kriegler li al., 1988; Moss et al., 1997). Txawm hais tias nws tau pom tias DOCAsalt ntshav siab tau tiv thaiv los ntawm TACE knockdown nyob rau hauv lub hlwb (Xia li al., 2013), nws txoj cai muaj tseeb hauvraumthaum lub sij hawm noj tshuaj HS ntev tsis tau txhais meej meej. Kev tshawb nrhiav ntxiv yuav xav tau los daws cov ntaub so ntswg. Txawm li cas los xij, nws tuaj yeem ntxiv rau ntawm no tias TNF- exerts nws cov lus teb biologic los ntawm kev sib cuam tshuam nrog ob lub xov tooj ntawm tes receptors, TNF- receptor hom 1 (TNFR1) thiab hom 2 (TNFR2), uas yog qhov sib txawv thiab tswj hwm hauvraum(Al- Lamki & Mayadas, 2015). Thaum mTNF- muaj qhov sib npaug sib luag rau ob qho tib si receptors, cov sTNF-muaj qhov siab tshaj plaws affinity los cuam tshuam nrog TNFR1 uas tsis muaj los yog tsawg affinity rau TNFR2 (Grell li al., 1998). Hauvraum, qhov twg TNFR2 ua lub luag haujlwm hauv lub raum mob (Singh li al., 2013), TNFR1 ua kom ua rau muaj cov nyhuv natriuretic los ntawm inhibiting tubular Na ntxiv rau kev thauj mus los thiab ENaC kev ua (Castillo et al., 2012) qhia lub luag haujlwm tiv thaiv TNF{{4 }} TNFR1 axis thaum lub sij hawm noj HS ntev. Hauv qhov xwm txheej zoo li no, qhov nce hauv plasma TNF- (sTNF- ) hauv cov lus teb rau kev noj tshuaj HS ntev uas tau pom nyob rau hauv txoj kev tshawb no yuav muaj lub luag haujlwm tseem ceeb hauv kev tswj hwm sodium homeostasis dua li cov ntaub so ntswg hloov pauv hauv lub raum hauv qib TNF- (mTNF-). Nws yog qhov nthuav kom nco ntsoov tias qhov kev noj haus HS ntev npaum li cas txo cov ntshav ntshav ntawm IL-6 hauv txoj kev tshawb no (Daim duab 3a), qhia txog lub luag haujlwm sib txawv ntawm cov cytokines pro-inflammatory hauv kev tswj hwm sodium homeostasis. Yog li ntawd, nws zoo nkaus li tias thaum TNF- raug tso tawm rau hauv kev ncig los ntawm lub tshuab ua haujlwm mononuclear phagocyte system (MPS) los ntawm osmoregulatory mechanism thaum lub sij hawm HS tau txais, kev tso tawm ntawm IL-6 zoo li feem ntau nyob ntawm lwm yam xws li nce renin- angiotensin system (Chamarthi li al., 2011). Thaum lub sij hawm HS noj ib leeg, renin-angiotensin system yog downregulated, thiab yog li, nws cuam tshuam rau IL -6 tsim yog tshwm sim nyob rau hauv ob qho tib si plasma thiab lub raum concentration ntawm no cytokine. Kev sim ntxiv yuav tsum tau ua kom nkag siab ntau ntxiv txog kev tswj hwm lub luag haujlwm ntawm IL -6 hauv lub raum excretory muaj nuj nqi.

Hauv txoj kev tshawb fawb tam sim no, nws tau sau tseg tias eNOSKO nas tau muab NS noj zaub mov muaj qib siab dua ntawm lub raum tubulointerstitial fibrosis, tab sis tsis yog glomerular sclerosis, piv rau WT nas. Cov kev hloov no zoo li yuav txo qis hauv eNOSKO nas muab HS noj zaub mov, qhia txog qhov cuam tshuam ntawm kev txo qis ntawm pro-inflammatory cytokines (TNF- thiab IL-6) hauv lub raum cov ntaub so ntswg. Xws li qhov txo qis ntawm lub raum raug mob hauv HS-fed eNOSKO nas zoo li tsis sib haum nrog qhov kev tshawb pom hauv lwm txoj kev tshawb fawb hauv eNOSKO nas tau tshaj tawm ua ntej (Daumerie li al., 2010) uas tsis ntsuas lub raum cytokine qib. Txawm li cas los xij, qhov sib txawv ntawm lub raum raug mob los ntawm txoj kev tshawb no (Daumerie et al., 2010) piv rau qhov kev tshawb fawb tam sim no tuaj yeem cuam tshuam rau cov hnub nyoog nas (kev sim pib ntawm 6 lub hlis vs. 2 lub hlis hnub nyoog) nrog rau qhov siab. cov ntsiab lus ntsev (8 feem pua ​​​​vs. 4 feem pua ​​NaCl) hauv cov khoom noj uas tau muab rau lub sijhawm ntev (8 lub lis piam vs 2 lub lis piam). Cov txiaj ntsig hauv txoj kev tshawb fawb tam sim no qhia tau hais tias, tsawg kawg hauv lub sijhawm luv, HS kev noj haus tau txo qis lub raum cytokine qib uas pab txo qis rau lub raum raug mob thaum ntxov.

Lub physiological lossis pathophysiological qhov tseem ceeb ntawm cov kev tshawb pom no uas HS tau txais tsawg kawg 2 lub lis piam ua rau txo qis hauv lub raum ntawm cov cytokines pro-inflammatory, tej zaum yuav yog qhov tseem ceeb rau kev tshawb nrhiav ntxiv. Txawm hais tias pro-inflammatory cytokine, tshwj xeeb tshaj yog TNF- tau cuam tshuam rau kev txhim kho ntsev rhiab heev thiab kub siab (Majid li al., 2015; Rodríguez-Iturbe li al., 2014; Schiffrin, 2013), kev tshawb fawb ua ntej hauv peb lub chaw kuaj mob (Shahid et al. al., 2008) qhia tau hais tias TNF- kev tswj hwm ua rau cov lus teb diuretic thiab natriuretic uas qhia tias TNF- ua lub luag haujlwm tiv thaiv hauv kev sib kho ntawm ntsev-sensitive hypertension. Yog li, kev txo qis hauv lub raum ntawm cov cytokines pro-inflammatory xws li TNF- hauv kev teb rau HS noj rau 2 lub lis piam tuaj yeem txhim kho cov tubular reabsorptive muaj nuj nqi uas tuaj yeem pab txhawb nqa sodium hauv lub cev. Xws li khaws cia ntawm sodium tuaj yeem txhim kho manifold nyob rau hauv cov xwm txheej ntawm oxidative kev nyuaj siab vim tsis muaj qhov tsis txaus. Ntsev rhiab heev pom nyob rau hauv eNOSKO nas tuaj yeem tshwm sim los ntawm kev txhim kho sodium retention nyob rau hauv cov xwm txheej ntawm oxidative kev nyuaj siab raws li peb tau pom ua ntej (Kopkan li al., 2010). Cov kev tshawb fawb ntxiv tuaj yeem xav kom nkag siab txog kev sib raug zoo ntawm kev sib raug zoo ntawm kev txo qis hauv lub raum ntawm cov cytokines pro-inflammatory cytokines thiab induction ntawm kub siab nyob rau hauv cov lus teb rau kev noj HS ntev. Txawm li cas los xij, nws tsim nyog rau postulate tias qhov txo qis hauv lub raum theem ntawm TNF- thiab lwm yam cytokines pro-inflammatory cytokines pab txhawb sodium retention ua rau induction ntawm kub siab nyob rau hauv teb rau kev noj HS noj.

XAIV

Cov kev tshawb pom nyob rau hauv txoj kev tshawb fawb tam sim no qhia tau tias HS tau txais tsawg kawg yog muab rau lub sijhawm 2-lub lim tiam, feem ntau inhibits lub raum theem ntawm inflammatory cytokines nyob rau hauv zoo raws li nyob rau hauv eNOS tsis muaj tej yam kev mob. Zuag qhia tag nrho, cov ntaub ntawv no qhia tias HS kev noj haus feem ntau ua rau txo qis ntawm cov pro thiab tiv thaiv cytokines hauvraum.Xws li kev txo qis ntawm cytokine ntau lawm, tshwj xeeb tshaj yog TNF- thaum noj HS, tej zaum yuav yog ib qho tseem ceeb ntawm eNOS deficiency uas yuav txhim khu lub raum tubular sodium reabsorption ua rau ntsev-tso tseg, thiab yog li, pab txhawb txoj kev loj hlob ntawm ntsev-sensitive hypertension.