Differential Lipidomics Ntawm HK-2 Cells Thiab Exosomes Hauv Qab Glucose Stimulation

Abstract

Qhov txawv txav ntawm cov cellular lipid metabolism yog qhov tseem ceeb heev hauv qhov tshwm sim thiab kev loj hlob ntawm tus kab mob ntshav qab zib raum (DKD). Txawm li cas los xij, cov lipid muaj pes tsawg leeg thiab kev qhia sib txawv los ntawm cov kua nplaum siab ntawm lub raum tubular hlwb thiab lawv cov exosomes, uas yog ib feem tseem ceeb ntawm kev loj hlob ntawm DKD, feem ntau tsis paub. Hauv txoj kev tshawb no, raws li kev tsom xam lipid tsom xam los ntawm isotope labeling thiab tandem mass spectrometry, tag nrho ntawm 421 thiab 218 hom lipid tau suav hauv HK-2 hlwb thiab exosomes, raws li. Qhov tseem ceeb tshaj, cov txiaj ntsig tau pom tias GM3 d18:1/22:0, GM3 d18:1/16:0, GM3 d18:0/16:0, GM3 d18: 1 / 22: 1 tau nce ntxiv, thaum LPE18: 1, LPE, CL66: 4 (16: 1), BMP36: 3, CL70: 7 (16: 1), CL74: 8 (16 :1) tau txo qis hauv cov piam thaj ntau ntxiv HK{{4{{9{98}}}}}} hlwb. Kuj, PG36:1, FFA22:5, PC38:3, SM d18:1/16:1, CE-16:1, CE-18:3, CE-20:5, thiab CE-22:6 tau nce ntxiv, thaum GM3 d18:1/24:1, GM3 tau txo qis hauv exosomes secreted los ntawm cov piam thaj-stimulated HK-2 hlwb. Tsis tas li ntawd, TAG, PC, thiab CL tau txo qis hauv exosomes piv nrog HK-2 hlwb, thiab LPA18:2, LPI22:5, PG32:2, FFA16:1, GM3 d18:1/18:1 , GM3 d18:1/2{100}}:1, GM3 d18:0/20:0, PC40:6p, TAG52:1(18:1), TAG52:0(18:0), CE{101 }}:5, CE-20:4, CE{105}}:6 tsuas yog pom hauv exosomes. Tsis tas li ntawd, qhov kev qhia ntawm PI4P hauv HK{108}} cov hlwb txo qis hauv cov piam thaj siab. Cov ntaub ntawv no yuav muaj txiaj ntsig los muab lub hom phiaj tshiab rau kev tshawb nrhiav cov txheej txheem ntawm DKD.

Ntsiab lus

Lub hom phiaj lipidomics; Lub raum tubules; Exosomes; Mob raum mob ntshav qab zib.

Nyem qhov no mus yuav Cistanche ntxiv los tiv thaiv lub raum

Taw qhia

Lipid metabolic ntshawv siab koom nrog qhov tshwm sim thiab kev loj hlob ntawm tus kab mob raum ntshav qab zib (DKD) [1, 2]. Kev hloov pauv hauv cellular lipid muaj pes tsawg leeg nyob rau hauv lub raum nyob rau hauv lub xeev siab qabzib tuaj yeem ua rau muaj kev nce hauv cov pa oxygen ntau lawm, mitochondrial tsis ua haujlwm, thiab txawm tias induce apoptosis [3, 4, 5, 6]. Txawm hais tias ntau tus kws tshawb fawb tau lees paub lub luag haujlwm tseem ceeb ntawm lub raum tubules hauv kev loj hlob thiab kev loj hlob ntawm DKD, ob peb txoj kev tshawb fawb tau hais txog tag nrho cov lipid qhia hauv lub raum tubules lossis cov kev hloov pauv hauv cov piam thaj siab [7, 8]. Yog li ntawd, nws yog ib qho tseem ceeb heev kom tau txais cov ntaub ntawv qhia txog lipids nyob rau hauv lub raum tubules, yog li ntxiv elucidating lub luag hauj lwm ntawm lipids nyob rau hauv lub pathogenesis ntawm DKD.

Nyob rau xyoo tas los no, lub luag haujlwm ntawm exosomes hauv kev txhim kho ntawm DKD tau nyiam ntau ntxiv. Exosomes uas tuaj yeem koom tes nrog autophagy-lysosome txoj hauv kev los txo cov kev ntxhov siab hauv lub cev muaj lub luag haujlwm tseem ceeb hauv kev tswj hwm cellular homeostasis [9, 10]. Lub caij no, lawv ua raws li cov neeg nqa khoom thauj khoom thiab cov ntaub ntawv sib txuas lus ntawm cov hlwb [11]. Cov kev tshawb fawb tsis ntev los no tau qhia tias kev sib tham ntawm cov tubular hlwb thiab lwm lub raum cell hom nyob rau hauv nephron tej zaum yuav muaj lub luag haujlwm tseem ceeb hauv kev kis tus kab mob [12, 13]. Txawm hais tias lipid muaj pes tsawg leeg ntawm exosomes tuaj yeem cuam tshuam rau lawv cov khoom, nws tau kawm txog qhov tsawg dua piv nrog cov xim ntawm cov protein thiab RNA [14]. Txij li thaum exosomes secreted los ntawm cov hlwb sib txawv hauv lipid hom, cov ntsiab lus, thiab kev ua haujlwm, qhia meej txog cov lipid muaj pes tsawg leeg thiab qhov sib txawv ntawm cov exosomes secreted los ntawm lub raum tubules nyob rau hauv lub xeev siab qabzib yuav muab lub hauv paus theoretical kom paub meej cov txheej txheem ntawm kev sib txuas lus exosomal txog raum tubules. .

Hauv qhov no, peb tau nthuav tawm qhov sib txawv lipidomic tsom xam ntawm lub raum tubules thiab lawv cov exosomes nyob rau hauv siab stimulation ntawm cov piam thaj thiab kuaj pom qhov txawv ntawm hom lipid cardiolipin (CL), monosialodihexosyl ganglioside (GM3), thiab cov roj cholesterol ester (CE). Peb kuj tau pib tshawb nrhiav lub luag haujlwm ntawm phosphoinositides (PIPs) hauv kev tsim tawm exosomal thiab tau soj ntsuam qhov nce ntawm tubular exosomes los ntawm glomerular mesangial cells (GMCs).

Cistanche ntxiv

Cov txheej txheem

1. Cell kab lis kev cai

HK-2 tib neeg lub raum proximal tubular epithelial cell kab tau yuav los ntawm Shanghai Institute for Biological Sciences Cell Resource Center. HK-2 hlwb raug coj los ua ke hauv cov piam thaj ib txwm muaj Dulbecco's Modified Eagle Medium/Nutrient Mixture F-12 (DMEM/F-12) uas tau ntxiv nrog 10 feem pua ​​​​fetal bovine serum (FBS; FSP500, ExCell). Cov piam thaj li qub DMEM/F-12 yog 1: 1 sib xyaw ntawm DMEM (11966025, Gibco) thiab Ham's F-12 (11765054, Gibco) uas muaj 5.56 mmol / L qabzib. Cov hlwb tau khaws cia ntawm 37 degree hauv 5 feem pua ​​CO2 incubator. Thaum 80 feem pua ​​​​ntawm cov confluence tau mus txog, cov hlwb tau sau los ntawm cov txheej txheem trypsin digestion thiab dhau ntawm qhov sib faib ntawm 1: 2. Cov kab mob kis tau raug coj los ntawm exosome-depleted FBS (CMS101.03, Cellmax) hauv 5.5 mmol·L{25}} qabzib raws li pawg tswj hwm dawb paug (NG) thiab 30 mmol·L{27}} qabzib raws li cov qabzib siab. pab pawg (HG) rau 48 teev.

2. Exosomes cais

Differential ultracentrifugation tau siv los rho tawm exosomes los ntawm HK {{0}} cell culture supernatants. Nyob rau hauv luv luv, HK-2 hlwb kab lis kev cai supernatants tau sau thiab sequentially centrifuged ntawm 2000 × g, 4 degree rau 10 min, thiab ces ntawm 10, 000 × g, 4 degree rau 30 min kom tshem tawm cov cell, cellular khib nyiab, thiab vesicles loj. Cov feem ntau supernatant tau lim siv 0.22-μm pore loj lim. Cov qauv lim tau ces raug ultracentrifugation ntawm 110,000 × g rau 75 min, ob zaug ntawm 4 degree rau pellet exosomes. Cov exosomes uas tau tshwm sim tau raug tshem tawm hauv ib qho me me ntawm phosphate buffer tov (PBS) thiab khaws cia ntawm -80 degree .

3. Nanoparticle tracking analysis (NTA)

Exosomes tau thawed hauv 25 degree dej da dej thiab muab tso rau hauv dej khov, tom qab ntawd lawv tau ncaj qha diluted nrog 1 × PBS rau kev kuaj pom nrog tunable resistive pulse sensing (TRPS). Kev soj ntsuam ntawm qhov loj me thiab qhov concentration ntawm exosomes tau ua tiav siv qNano Kub nrog Izon Control Suite 3.3.2 los ntawm Izon Science.

4. Protein kev txiav txim siab thiab western blotting

Kwv yees li. 100ul premixed cracking tov (RIPA cracking tov thiab 1 feem pua ​​PMSF inhibitor) tau ntxiv rau ntawm tes thiab exosome kuaj cais. Tom qab tawg rau 30min, lawv tau centrifuged ntawm 12, 000 × g, 4 degree rau 20min. Cov supernatant tau noj, thiab cov protein ntau tau txiav txim siab los ntawm kev siv bicinchoninic acid (BCA; Beyotime) cov khoom kuaj protein raws li cov chaw tsim khoom cov lus qhia. Bovine serum albumin tau siv los ua tus qauv protein.

Cov qauv ntawm cov protein tau muab faib los ntawm sodium-dodecyl-sulfate polyacrylamide gel electrophoresis (SDS-PAGE). Tom qab raug xa mus rau PVDF daim nyias nyias (Millipore), lawv tau tsim nrog ntau yam tshuaj tiv thaiv kab mob, anti-CD63 (EXOAB-CD63A-1, SBI; 1: 1000), anti-ALIX (YT6283, Immunoway; 1: 1000). ), anti-Calnexin (YT0613, Immunoway; 1: 1000), anti-P62 (P0067, Sigma; 1: 1000), thiab anti-LC3 (L7543, Sigma; 1: 1000), tom qab ntawd los ntawm tshis los tiv thaiv luav lossis nas. Ig Secondary Antibodies (180202-001, SBI; 1:5000). Cov qhab nia tshwj xeeb tau kuaj pom siv cov khoom siv hluav taws xob zoo dua qub (ECL; Beyotime) substrate. -actin (BM0627, Boster) tau siv los ua kev tswj hwm sab hauv. Tus txheeb ze band siv tau ntsuas los ntawm kev siv ImageJ software.

5. Transmission electron microscopy (TEM)

5ul coj mus kuaj tau muab tso rau hauv tooj liab net thiab incubated ntawm chav tsev kub rau 5min. Ib qho poob ntawm 2 feem pua ​​​​ntawm uranium peroxide acetate tau ntxiv rau hauv cov tooj liab thiab incubated ntawm chav tsev kub rau 1 min. Tom qab ntawd nws tau qhuav rau li 20min ntawm chav tsev kub thiab pom nyob rau hauv nano-transmission electron microscope (Tecnai G2 Spirit BioTwin, FEI).

Cistanche extract

6. Lub hom phiaj lipidomics

Lipids tau muab rho tawm los ntawm kev hloov kho ntawm Bligh thiab Dyer txoj kev [15]. Txhua qhov kev ntsuam xyuas lipidomic tau ua tiav ntawm Exion ultra-performance kua chromatography (UPLC) ua ke nrog SCIEX QTRAP 6500 PLUS system. UPLC-MS/MS kev soj ntsuam tau ua nyob rau hauv electrospray ionization (ESI) hom, nrog rau cov hauv qab no: ntaub thaiv npog roj=20, ion spray voltage=5,500 V , kub=400 degree , ion qhov chaw roj 1=35, thiab ion qhov chaw roj 2=35. Polar lipids tau muab sib cais siv Phenomenex Luna 3 µm ntawm silica kem (sab hauv kab 150 × 2.0 mm) nrog ob theem mobile: mobile theem A (chloroform: methanol: ammonium hydroxide, 89.5: 10: 0.5) thiab mobile theem B (chloroform: methanol: ammonium hydroxide: dej, 55: 39: 0.5: 5.5). Kev sib cais gradient tau ua raws li hauv qab no: qhov gradient tau khaws cia nrog 95 feem pua ​​​​A rau 5 feeb thiab tom qab ntawd txo qis mus rau 60 feem pua ​​​​hauv 7 min thiab tuav rau 4 min, tom qab ntawd nws tsis kam mus rau 30 feem pua ​​thiab ces tuav rau 15 min. Thaum kawg, thawj gradient tau thov thiab khaws cia rau 5 feeb. Mass spectrometric multi-react monitoring (MRM) tau tsim los rau ntau yam kev txheeb xyuas lipid thiab kev tsom xam ntau [16, 17]. Cov tib neeg polar lipid hom tau suav los ntawm kev xa mus rau cov qauv sab hauv ntawm tib chav lipid suav nrog PE-d31(16:0/18:1), DMPE; PG-d31(16:0/18:1), DMPG; PI-d31(16:0/18:1), di-C8- PI; PS-d31(16:{98}}/18:1), DMPS; PA-d31(16:{{1{{1{11{115}}}}8}}4}}/18:1), PA(17:{120}}/17:0); BMP-(14:0/14:0); LPE-17:1; LPI-17:1; LPS-17:1; LPA-17:0; GM3 d18:1/18:0-d3; d31-16:0, d8-20: 4; PC-d31(16:0/18:1), DMPC; SM d18:1/d31-16:0, SM d18:1/12:0; LPC{109}}:0; Cer d18:1-d7/15:0; Sph-d18:1/17:0; 22:1(3){124}}:1.

Glycerol lipids, suav nrog diacylglycerols (DAGs) thiab triacylglycerols (TAGs), tau suav nrog siv cov hloov pauv hloov pauv ntawm theem HPLC / MS / MS. Kev sib cais ntawm cov lipids nruab nrab tau ua tiav ntawm Phenomenex Kinetex-C18 2.6 µm kem (id 4.6x100 mm) siv lub sijhawm txawb uas muaj chloroform: methanol: {{19} }.1 M ammonium acetate (100: 100:4), ntawm tus nqi ntws ntawm 160 µl/min. d5-TAG (16:0)3; d5-TAG (14:0)3; d5-TAG (18:0)3 tau siv rau tus kheej TAG. d5-DAG (1,{27}}:0); d5-DAG (1,{31}}:1) tau siv los ua kom muaj DAG tus kheej raws li Neutral nco MS/MS thev naus laus zis.

Cov roj cholesterol dawb, sterols, thiab lawv cov esters raug tshuaj xyuas los ntawm HPLC-MS / MS, ua nyob rau hauv atmospheric siab tshuaj ionization (APCI) hom, nrog cov roj (cholesterol) -2, 2,3,4,4, 6-d6 Octadeconate; Cov roj (cholesterol)-26,26,26,27,27,27-d6 raws li cov qauv sab hauv.

Chloroform: methanol: (1:1), 2.4 M hydrochloric acid, thiab 0.25 M EDTA tau ntxiv rau cov qauv, ua raws li kev sib tsoo ntawm qhov kub thiab txias. Cov qauv yog centrifuged tom qab incubation rau 3 teev ntawm 4 degree, tom qab uas lub qis organic theem tau muab rho tawm. Chloroform tau ntxiv rau qhov thib ob extraction, tom qab ntawd ntxuav cov organic theem siv 1N hydrochloric acid: methanol (1: 1). Cov theem ntawm cov organic yog qhuav nyob rau hauv lub tshuab nqus tsev rotary concentrator. Raws li yav dhau los tau tshaj tawm, 40 feem pua ​​​​methylamine: dej: n-butanol: methanol (36:8:9:47) tau siv rau deacylation thiab tshuaj xyuas los ntawm Thermo ICS-5000 ion chromatography. PI4P, PI3P, PI4P, PI (3, 4) P2, PI (3, 5) P2, PI (4, 5) P2, thiab PI (3, 4,5) P3 tau siv los tsim ib qho kev ntsuas sab nraud rau ntau qhov tsom xam. Kev tshuaj xyuas tau txhawb nqa los ntawm Lipidall Technologies Company Limited.

7. Fluorescence microscopy

Txhawm rau pom qhov nce ntawm exosomes rau hauv GMCs (Sicencell), qhov sib npaug ntawm cov exosomes secreted los ntawm HK-2 hlwb nrog lossis tsis muaj cov piam thaj ntau ntxiv yog ob qho tib si sau nrog PKH67 (mini67, Sigma) raws li cov chaw tsim khoom cov lus qhia. Labeled exosomes tau coj mus kuaj nrog GMCs rau 24 teev. Confluent GMCs tau ntxuav peb zaug nrog PBS, tom qab ntawd cov hlwb raug kho nrog 4 feem pua ​​​​ntawm paraformaldehyde thiab khaws cia tawm ntawm lub teeb rau 30 feeb. Cells raug ntxuav peb zaug nrog PBS thiab stained nrog DAPI (28718903, Solarbio) rau 4-6min. Tom qab ntxuav peb zaug, confluent GMCs tau yees duab siv Nikon C2 confocal microscope.

8. Kev txheeb cais

GraphPad 7. Cov ntaub ntawv raug qhia raws li txhais tau tias ± SEM. Cov tub ntxhais kawm qhov t-test tau siv los sib piv ntawm ob pawg. P-tus nqi < 0.05 raug suav hais tias yog qhov tseem ceeb.

Cistanche capsules

Kev sib tham

Peb txoj kev tshawb fawb muab cov hom lipid sib txawv ntawm HK{{0}} cov hlwb thiab cov exosomes nyob rau hauv cov ntshav qabzib siab, uas tsis tau tshaj tawm yav dhau los. Interestingly, hom GM3, xws li GM3 d18:1/22:0, GM3 d18:1/16:0, GM3 d18:0/16:0 , GM3 d18: 1 / 22: 1 tau nce siab hauv cov piam thaj-stimulated HK-2 hlwb, thiab GM3 d18: 1 / 24: 1, GM3 tau txo qis hauv lawv cov exosomes. Tsis tas li ntawd, GM3 d18: 1 /18:1, GM3 d18:1/20:1, thiab GM3 d18:0/20:0 tsuas yog pom hauv exosomes. GM3 yog qhov dav dav dav dav ganglioside hauv lub cev, suav nrog cov piam thaj, galactose, thiab sialic acid uas ua los ntawm cov pab pawg sialic cuam tshuam nrog cov txheej txheem ceramide skeleton [18]. Nws koom nrog kev tswj hwm ntawm ntau yam ntawm tes ua haujlwm, suav nrog kev xa tawm kev hloov pauv, kev loj hlob, kev sib txawv, thiab apoptosis. Raws li cov kev tshawb fawb tsis ntev los no, cov ntshav concentration ntawm GM3 yog siab dua nyob rau hauv cov neeg mob uas muaj hom 2 mob ntshav qab zib, hyperlipidemia, thiab cov neeg mob rog rog, thaum GM3 tuaj yeem txhawb kev tshem tawm cov tshuaj insulin thiab txo cov teeb liab insulin [19]. Peb cov txiaj ntsig tau qhia txog qib siab ntawm txhua qhov sib txawv qhia GM3 hom hauv HK-2 hlwb nyob rau hauv cov kua nplaum siab, tshwj xeeb tshaj yog qhov nce ntawm GM3 nrog cov saw hlau sab nraud (22: 0) uas yog qhov pom tseeb tshaj plaws. Zhang et al. [20] kuj tau pom GM3 d18: 1 / 22: 0 qhia nyob rau hauv lub raum cortex ntawm cov kab mob nephropathy nas, uas yog raws li peb cov txiaj ntsig. Kev tshawb fawb ntawm streptozotocin-induced type 2 diabetes nas ua los ntawm Anela li al. [21] pom tsis hloov GM3 nyob rau hauv lub glomeruli tab sis ho nce GM3 qhia nyob rau hauv lub raum tubules, uas yog nyob rau hauv txoj kab nrog peb nrhiav tau ntawm GM3 hloov nyob rau hauv lub raum tubules. Kwak et al. [22] pom GM3 txo qis hauv cov ntsiab lus glomerular ntawm streptozotocin-induced ntshav qab zib nas, ua raws li qhov poob ntawm cov nqi xaiv pom teeb meem hauv glomeruli. Nws yog ib qho tseem ceeb uas yuav tsum nco ntsoov tias GM3 lub luag haujlwm hauv glomeruli thiab lub raum tubules yuav txawv. GM3 kuj yog ib feem tseem ceeb ntawm lipid raft [23]. Kev hloov pauv hauv GM3 qib qhia hauv lipid rafts hloov cov dej num ntawm Na ntxiv -glucose cotransporter hom 2 (SGLT2) thiab lub raum Na plus / K ntxiv / Cl- cotransporter, uas yog ob qho tib si lipid raft-dependent [24]. Qee cov kws tshawb fawb tau xav tias qhov siab dua ntawm GM3 nce SGLT2 thiab Na plus / K ntxiv / Cl- cotransporter kev ua ub no, ua rau muaj zog tubular reabsorption thiab efferent arteriole hydrostatic siab thiab tom qab ntawd ua rau lub raum tubular cell raug mob los ntawm kev puas tsuaj rau tubuloglomerular tshuav nyiaj li cas. Kumari et al. [25] pom tias GM3 hauv cov zis exosomes ntawm cov neeg mob tau pom qhov sib txawv ntawm DKD thiab ntshav qab zib mellitus. Ua ke, lub luag haujlwm ntawm GM3 lub siab qhia nyob rau hauv cov piam thaj siab hauv lub raum tubules yuav tsum tau hais los ntawm kev tshawb fawb yav tom ntej.

Qhov tseem ceeb, peb cov txiaj ntsig kuj tau pom tias qee hom CL, suav nrog CL66: 4 (16: 1), CL70: 7 (16: 1), thiab CL74: 8 (16: 1) tau txo qis hauv cov piam thaj-stimulated HK{ {13}} hlwb. Piv nrog HK-2 hlwb, CL tau txo qis hauv cov exosomes. CL yog nyob yuav luag tag nrho hauv mitochondrial puab daim nyias nyias thiab yog ib feem tseem ceeb ntawm kev tswj cov qauv mitochondrial. Nws muaj lub luag haujlwm tseem ceeb hauv electron conduction, adenosine triphosphate ntau lawm, lub zog metabolism, thiab apoptosis [26, 27]. Hauv kev mob ntshav qab zib mellitus, mitochondrial morphological hloov pauv thiab ua haujlwm tsis zoo feem ntau nrog rau kev hloov pauv CL [28,29]. Lub proximal lub raum tubular epithelial hlwb xav tau ATP txaus los tswj cov kev thauj mus los thiab rov qab nqus ntawm ntau yam khoom. Raws li lub zog siv hluav taws xob siab, lub raum lub raum tubular epithelial hlwb yog nplua nuj nyob rau hauv mitochondria [30]. Mitochondrial fragmentation tau pom nyob rau hauv proximal tubular epithelial hlwb hauv cov ntshav qab zib thaum ntxov. Zhang et al. [31] pom tias CL yog qhov tseem ceeb rau kev tswj cov tubular mitochondrial muaj nuj nqi hauv cov nas uas muaj ntshav qab zib hom 1. Peb cov txiaj ntsig yuav muab lub hauv paus tshiab rau kev kawm txog kev raug mob mitochondrial mechanism ntawm DKD.

Exosomes muaj lub luag haujlwm tseem ceeb hauv kev tswj hwm cellular homeostasis los ntawm kev koom tes nrog txoj kev autophagy-lysosome. Cov kev tshawb fawb yav dhau los tau pom tias kev hloov pauv hauv PIPs qhia qib tuaj yeem cuam tshuam rau ob qho tib si exosome secretion thiab cellular autophagy los ntawm kev tswj hwm kev hloov pauv ntawm MVBs [32, 33, 34]. Nina et al. [35] pom tias nyob rau hauv PC-3 hlwb, inhibition ntawm PIKfyve, uas yog substrate yog PI3P, ua rau kom lub secretion ntawm exosomes thiab induced secretory autophagy, yog li qhia tau hais tias cov kev sib raug zoo los ntawm PIPs. Raws li cov lus hais saum toj no, peb tau pom cov kev hloov pauv hauv PIPs, exosomal ntau lawm, thiab autophagy hauv cov piam thaj-stimulated HK-2 hlwb. Peb cov txiaj ntsig tau pom tias cov piam thaj siab tau txhawb nqa ob qho tib si tso tawm ntawm exosomes thiab ua kom autophagy hauv HK{11}} hlwb, thaum muaj qhov txo qis hauv PI4P qhia. Cov kev tshawb fawb ntxiv yog tsim nyog los tshawb xyuas cov txheej txheem exosome-autophagy kev tswj hwm ntawm PIPs koom nrog kev txhawb nqa cov piam thaj siab.

Raws li tus neeg xa xov liaison, exosomes muaj lub luag haujlwm tseem ceeb hauv intracellular signaling thiab ua haujlwm hloov pauv hauv kev txhim kho DKD [36, 37, 38]. Peb fluorescently sau npe HK-2 hlwb-tso tawm exosomes thiab pom lawv cov paracrine uptake los ntawm GMCs. Piv nrog rau lawv niam lub hlwb, exosomes muaj ib qho tshwj xeeb muaj pes tsawg leeg, uas ua rau lawv muaj kev ruaj khov thiab ua haujlwm zoo vim muaj cov roj cholesterol ntau, sphingolipids, saturated phospholipids, thiab lipid raft domains [39]. Txawm tias me me hloov hauv lipid muaj pes tsawg leeg tuaj yeem cuam tshuam cov khoom ntawm daim nyias nyias, yog li muaj txiaj ntsig zoo rau nws txoj haujlwm. Hauv txoj kev tshawb no, peb pom tias TAG, PC, thiab CL tau txo qis hauv exosomes piv nrog HK-2 niam hlwb, thaum 13 hom lipids suav nrog LPA18:2, LPI22:5, PG32:2, FFA16: 1, GM3 d18:1/18:1, GM3 d18:1/20:1, GM3 d18:{{30}}/20:0, PC40: 6p, TAG52:1(18:1), TAG52:0(18:0), CE-20:5, CE-20:4, CE-22:6 tsuas yog kuaj pom hauv exosomes. Cov hauj lwm nthuav tawm no muab lub hauv paus rau kev tshuaj xyuas ntxiv ntawm paracrine lub luag haujlwm ntawm exosomes tso tawm los ntawm tubular hlwb.

Cistanche hmoov

Cov lus xaus

Hauv kev xaus, peb txoj kev tshawb fawb tau pom qhov tseem ceeb ntawm kev soj ntsuam lipidomics hauv kev muab lub hom phiaj tshiab los tshawb txog cov kab mob ntawm DKD. Cov kev tshawb fawb ntxiv txog kev ua haujlwm ntawm qhov sib txawv ntawm cov lipids thiab cov kev tshawb fawb txog lipid ntawm lwm cov hlwb hauv nephron thiab lawv cov exosomes tuaj yeem muab qhov kev xav tshiab uas yuav ntxiv dag zog rau cov txheej txheem ntawm DKD.

Cov ntaub ntawv

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34. Tore Skotland, Nina P. Hessvik, Kirsten Sandvig, Alicia Llorente. Exosomal lipid muaj pes tsawg leeg thiab lub luag haujlwm ntawm ether lipids thiab phosphoinositides hauv exosome biology. J Lipid Res. 2019; 60(1): 9-18.

35. Nina Pettersen Hessvik, Anders Øverbye, Andreas Brech, Maria Lyngaas Torgersen, Ida Seim Jakobsen, Kirsten Sandvig. ua al. PIKfyve inhibition nce exosome tso tawm thiab induces secretory autophagy. Cell Mol Lub Neej Sci. 2016; 73: 3 :ua.

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38. Xiao-ming Wu, Yan-bin Gao, Fang-qiang Cui, Na Zhang. Exosomes los ntawm cov piam thaj siab kho glomerular endothelial hlwb ua kom cov hlwb mesangial los txhawb lub raum fibrosis. Biol Qhib. 2016; 5(4): 484-91.

39. Tore Skotland, Kirsten Sandvig, Alicia Llorente. Lipids hauv exosomes: Kev paub tam sim no thiab txoj hauv kev rau pem hauv ntej. Prog Lipid Res. 2017; 66: 3 :ua.

Weidong Wang, Tingting Li, Zhijie Li, Hongmiao Wang, Xiaodan Liu

Department of Nephrology, Thawj Tsev Kho Mob ntawm Tuam Tshoj Medical University, 155 North Nanjing Street, Shenyang, Liaoning, PR Tuam Tshoj, 110001