Pem hauv ntej hlwb. 2018; 9: 81.
Luam tawm hauv internet 2018 Mar 12. doi: 10.3389 / fpsyt.2018.00081
PMCID: PMC5858605
PMID: 29593587
Minho Lee,1,† Hyeyoung Cho,2,3,† Seung Hyun Jung,2,4 Seon-Hee Yim,2 Sung-Min Cho,2,3 Ji-Won Chun,5 Soo-Hyun Paik,5 Yae Eun Park,6,7 Dong Huey Cheon,7,8 Ji Eun Lee,7 Jung-Seok Choi,9 Dai-Jin Kim,5,* thiab Yeun-Jun Chung1,2,3,*
Abstract
Tom qab
Hom kev siv internet thiab hauv internet ua si yog ib qho kev puas siab puas ntsws tsis hais ua rau hauv Internet gaming teeb meem (IGD). Altered microRNA (miRNA) kev qhia tau raug tshaj tawm hauv cov ntshav thiab lub hlwb ntawm cov neeg mob uas muaj kev puas siab puas ntsws thiab pom tias biomarkers. Txawm li cas los, tsis muaj cov ntaub ntawv qhia txog ntshav miRNA profiles hauv IGD.
txoj kev
Mus nrhiav tau IGD-txuas miRNAs, peb tau soj ntsuam cov kev qhia txog cov khoom ntawm 51 25 (26 IGD thiab 36 ntawm kev tswj) siv cov TaqMan Low Density miRNA Array. Rau validation, peb ua tau ntau dua Reversed PCR nrog 20 independent kuaj (16 IGD thiab XNUMX tswj).
tau
Los ntawm kev tshawb nrhiav thiab kev ywj siab, peb pom tias peb miRNAs (hsa-miR-200c-3p, hsa-miR-26b-5p, hsa-miR-652-3p) uas tau txo qhov teeb meem hauv pawg IGD. Cov neeg uas muaj tag nrho peb cov kev ntsuas miRNA hloov tau muaj feem ntau ntawm cov kab mob IGD dua cov uas tsis muaj kev hloov [kev sib txawv ntawm qhov txawv (LOS YOG) 22, 95% CI 2.29-211.11], thiab cov ORS ntau ntxiv nrog rau cov kev hloov kho miRNAs. Lub hom phiaj ntawm lub hom phiaj ntawm peb lub miRNAs tau txuam nrog txoj kev ua neural. Peb tau tshawb cov protein ntau ntawm peb lub hauv paus ntawm cov noob qis thiab paub tseeb tias qhov qhia ntawm GABRB2 thiab DPYSL2 tau yooj yim dua hauv pawg IGD.
xaus
Peb pom tias cov kab lus ntawm hsa-miR-200c-3p, hsa-miR-26b-5p, thiab hsa-miR-652-3p raug txo nyob hauv cov neeg mob IGD. Peb cov qhab-nees yuav pabcuam kom nkag siab txog qhov kev mob ntsws ntawm IGD.
Introduction
Kev siv hom kev siv Internet thiab Internet-based games tsis yog ib qho tshwm sim hauv lub teb chaws uas muaj kev siv internet yooj yim heev, tiam sis muaj kev puas siab puas ntsws tsis hais ua rau hauv Internet gaming teeb meem (IGD)1-3). Raws li cov ntawv ceeb toom epidemiological, feem ntau ntawm IGD cov me nyuam hluas sib txawv thoob lub teb chaws, xws li 0.8 rau 26.7% (4). Tshwj xeeb tshaj yog, cov kev tshawb fawb pom tias muaj ntau tshaj li ntawm 10% cov tub ntxhais hluas nyob rau ntau lub teb chaws Asia xws li South Kauslim Tebchaws, Tuam Tshoj, Taiwan, Hong Kong, thiab Singapore (4). IGD yog txuam nrog kev puas hauv cognition, psycho-social relationships, thiab lub neej txhua hnub; Piv txwv, poob qib kev kawm lossis kev ua haujlwm (4-7). IGD yog tam sim no muaj nyob rau hauv Tshooj III (Cov Kws Tshaj Lij rau Txoj Kev Kawm Tshaj Lij) ntawm txoj cai thib tsib ntawm Txoj Cai Diagnostic thiab Cov Ntawv Cuam Tshuam ntawm Kev Xiam Hlwb ntawm Kev Nyuaj Siab (DSM-V) (8). Txawm li cas los xij, qhov tseem ceeb ntawm nws qhov kev mob-kev tseem ceeb, me ntsis yog paub txog molecular caj tshuab tom qab IGD.
Tsis ntev los no ntau-kev sib tw kev tshawb fawb tau pom tias qhov caj ces keeb kwm yav dhau los IGD (9, 10). Vink li al. tshawb xyuas tib neeg kev sib txawv hauv kev siv Internet nrog 5,247 monozygotic thiab cov menyuam yau hauv cov tub ntxhais hluas ntxhais ntxaib hauv lub Tebchaws Twin Register thiab qhia tias 48% ntawm qhov txawv tau piav qhia los ntawm cov caj ces (9). Li li al. pom 825 khub ntawm Suav cov tub ntxhais hluas ntxaib thiab qhia tias cov caj ces piav qhia 58-66% ntawm qhov sib txawv (10). Raws li, polymorphisms ntawm cov noob koom hauv neurotransmission, cognition, thiab xim xws li dopamine receptor D2 noob (DRD2), catecholamine-O-methyltransferase noob (COMT), serotonin transporter noob (5HTTLPR), thiab cholinergic receptor nicotinic alpha 4 noob (CHRNA4) tau raug tshaj tawm tias yuav raug cuam tshuam nrog kev tiv thaiv hauv Internet (11-13). Tsis ntev los no, Kim et al. ntsuam xyuas cov kab txawv ntawm ntau tshaj 100 tus neeg sib tw noob nrog ntau lawm, kev ua, thiab cov metabolism hauv neurotransmitters los ntawm lwm tiam neeg qhov kev soj ntsuam thiab qhia tias rs2229910 ntawm NTRK3 noob yog txuam nrog IGD (14).
Ntxiv rau cov caj ces, nws tseem paub zoo tias neurobehavioral phenotypes yog epigenetically tswj los ntawm non-coding RNAs xws li microRNAs (miRNAs) (15, 16). miRNAs yog cov me-uas tsis muaj kev sib tw RNA molecules (kwv yees 20-23 nucleotides nyob ntev), uas cuam tshuam tswj cov protein-coding noob by degrading mRNAs thiab ua si lub luag haujlwm tseem ceeb hauv txoj kev mob ntawm cov kab mob ntau yam (cov kab mob pathophysiological17). Cov pov thawj ntawm cov pov thawj tau pom tias cov miRNAs muaj ntau nyob rau hauv tib neeg lub hauv nruab nrab hauv lub paj hlwb (CNS) thiab ua kom zoo qhib cov qib theem ntawm lawv lub hom phiaj, uas muaj kev koom tes hauv kev loj hlob thiab kev loj hlob ntawm CNS system (15). Tseeb, tsis ntev los no cov kev tshawb fawb muaj ices tias miRNA qhia profiles yog kho hauv lub hlwb cov nqaij mob ntawm cov neeg mob uas muaj kev puas hlwb, tawm tswv yim tias lawv cov ntaub ntawv qhia tau yog biomarkers rau cov kev puas siab puas ntsws (15, 16, 18). Piv txwv li, los ntawm kev tshawb nrhiav qhov tseeb, Lopez li al. qhia tias kev qhia ntawm miR-1202, uas tswj cov kev qhia ntawm metabotropic glutamate receptor-4 noob thiab kwv yees cov lus teb rau antidepressant, yog downregulated nyob prefrontal cortex cov nqaij ntawm cov neeg mob nyhav loj (19). Nyob rau hauv cov nqe lus ntawm biomarker kev soj ntsuam, txoj kev qhia no muaj meej meej vim hais tias ua lub zog ntawm nqaij tawv nqaij CNS rau kev kuaj xyuas yog tsis yooj yim sua. Txij thaum miRNAs tuaj yeem kuaj tau cov ntshav (ntshav lossis ntshav dej), circulating miRNAs muaj meej kom zoo dua li cov uas tsis yog tus neeg mob biomarkers hauv kev puas siab ntsws (neuropsychiatric disorders). Txawm li cas los, hnub tim, tsis muaj kev tshawb fawb txog circulating miRNA profiles hauv IGD. Zoo dua kev to taub txog kev ncig ntawm mirNA qhia txog kev nkag siab tau pab qhia ntxiv txog txoj kev txhim kho IGD thiab pab txhawb kev txhais lus.
Nyob rau hauv txoj kev tshawb no, peb xav kom txheeb xyuas cov tshuaj miRNA uas yog IGD-txheeb los ntawm kev soj ntsuam sib txawv ntawm cov ntshav mlRNAs ntawm IGD thiab tswj hwm pawg thiab tshawb txog lawv qhov kev xeeb menyuam.
Ntaub ntawv thiab kev
Kawm Yam Kawm
Peb tau soj ntsuam 3,166 cov hluas (hnub nyoog 12-18 xyoo) uas siv DSM-V IG tw. Ntawm lawv, 251 (168 Caug thiab 83 pojniam) raug kuaj tau raws li IGD raws li DSM-V cov cai (8). Ib qho tag nrho ntawm 91 cov neeg (49 IGDs thiab 42 tswj) tau txais kev tso cai pom zoo rau txoj kev tshawb no. Ntawm lawv, plaub tus neeg tau raug tshem tawm raws li kev cais tawm. Thaum kawg, 87 cov neeg (45 IGD cov ntsiab lus thiab 42 cov neeg tswj kev noj qab nyob zoo) tau koom rau txoj kev tshawb no. Ntawm lawv, 51 cov neeg koom (25 IGD cov neeg mob thiab 26 tswj) tau nrhiav ua cov kev nrhiav pom los ntawm 2014 rau 2016. Lwm tus 36 cov neeg tuaj koom (20 IGD cov neeg mob thiab 16 tswj) tau nrhiav ua tus neeg sab nraud validation los ntawm 2016. Tag nrho cov neeg koom ua ke tau yog cov neeg Korean, Seoul St. Mary lub Tsev Kho Mob (Seoul, South Kauslim) thiab Seoul National University Boramae Tsev Kho Mob (Seoul, Kaus Lim Kauslim Teb). Tag nrho cov neeg tau tuaj yeem muaj kev sib tham los ntawm ib tug kws kho mob raws li Kiddie Kiddie Schedule rau Affective Disorders thiab Schizophrenia (K-SADS-PL) (20). Tag nrho cov neeg koom ua tiav Lub Nroj Tsim thiab Cov Ntawv Kawm cov ntawv raws li Kauslim-Wechsler Kev Ntsuas Ntawm Cov Me Nyuam rau Cov Me Nyuam, 4th tsab (K-WISC-IV) (21). Impulsiveness raug soj ntsuam los ntawm Barratt Impulsiveness Nplai (BIS) (22). Cwj Pwm Cuam Tshuam Txheej Txheem (BInS) thiab Cwj Pwm Cuam Tshuam Txheej (BAS) cov ntsuas tau los ntsuas tus kheej qhov dav (23). Cov kev cais tawm ntawm cov teebmeem nrog yav dhau los lossis tam sim no kev mob nkeeg (xws li, ntshav qab zib mellitus), teeb meem txog kev puas hlwb (xws li qaug dab peg, mob taub hau), kev puas hlwb (xws li kev nyuaj siab loj, kev ntxhov siab, kev ntxhov siab), kev ntxhov siab, , haus luam yeeb, cannabis, cawv). Cov ntsiab lus ntawm cov kev kawm sib txawv yog sau ua ke hauv Cov Ncauj Lus Table1.1. Txoj kev tshawb no tau pom zoo los ntawm Institutional Review Board ntawm Catholic University Kawm Kev Kawm Txuj Ci Qib Kauslim (MC16SISI0120). Tag nrho cov neeg tuaj koom thiab lawv niam lawv txiv tau sau daim ntawv pom zoo.
rooj 1
Cov yam ntxwv ntawm cov qauv kawm.
| Discovery | Validation | Ua ke | |||||||
|---|---|---|---|---|---|---|---|---|---|
| Control | IGD | P-peem | Control | IGD | P-peem | Control | IGD | P-peem | |
| N | 26 | 25 | 16 | 20 | 42 | 45 | |||
| Hnub nyoog (xyoo) | |||||||||
| Median (min-max) | 13 (12 - 17) | 13 (12 - 15) | 0.759 | 15 (13 - 18) | 14.5 (12 - 18) | 0.628 | 14 (12 - 18) | 14 (12 - 18) | 0.509 |
| Txhua lub sijhawm ua si hauv Internet (h) | |||||||||
| Median (min-max) | 5.25 (2 - 17) | 18 (6 - 46) | 1.27E-6a | 5.5 (2 - 23) | 8 (1 - 112) | 0.374 | 5.5 (2 - 23) | 14 (1 - 112) | 1.63E-5a |
| Cov nyiaj tau los hauv tsev neeg txhua hli (lab ntawm KRW) | |||||||||
| Median (min-max) | 5 (1 - 9) | 3 (1 - 9) | 0.588 | 4 (4 - 4) | 2 (2 - 2) | 1.000 | 5 (1 - 9) | 3 (1 - 9) | 0.460 |
| Kev kawm ntawv (xyoo) | |||||||||
| Median (min-max) | 8 (7 - 9) | 8 (7 - 9) | 0.584 | 12 (12 - 12) | 6 (6 - 13) | 0.305 | 8 (7 - 12) | 8 (6 - 13) | 0.269 |
| K-WISC: thaiv qauv | |||||||||
| Median (min-max) | 10.5 (4 - 17) | 10 (4 - 16) | 0.544 | 10 (3 - 16) | 12.5 (4 - 15) | 0.125 | 10 (3 - 17) | 11 (4 - 16) | 0.598 |
| K-WISC: cov ntsiab lus | |||||||||
| Median (min-max) | 9 (5 - 17) | 7 (5 - 13) | 0.174 | 9.5 (8 - 15) | 11.5 (5 - 15) | 0.595 | 9 (5 - 17) | 9 (5 - 15) | 0.527 |
| KS | |||||||||
| Median (min-max) | 24 (17 - 36) | 37 (22 - 51) | 3.81E-6a | 29 (17 - 34) | 59 (22 - 108) | 1.2E-5a | 25 (17 - 36) | 40 (22 - 108) | 2.05E-10a |
| BIS | |||||||||
| Median (min-max) | 63 (35 - 75) | 67.5 (45 - 81) | 0.080 | 61 (45 - 79) | 63 (32 - 82) | 0.835 | 62 (35 - 79) | 65 (32 - 82) | 0.240 |
| QIS | |||||||||
| Median (min-max) | 31 (15 - 40) | 31 (13 - 51) | 0.558 | 36.5 (22 - 48) | 34 (27 - 52) | 1.000 | 32 (15 - 48) | 34 (13 - 52) | 0.637 |
| BInS | |||||||||
| Median (min-max) | 18 (10 - 26) | 17.5 (13 - 27) | 0.642 | 18.5 (12 - 25) | 20 (13 - 21) | 0.138 | 18 (10 - 26) | 19 (13 - 27) | 0.302 |
IGD, Cov neeg mob hauv Internet gaming; KS, Kauslim Teb Internet Kev Lom Zem; BIS, Barratt Impulsivity Nplai; BAS, Kev Cuam Tshuam Ua Haujlwm; BInS, Kev Coj Tus Cwj Pwm Zoo; KRW, Kauslim Kauslim.
aP <0.05 (Mann – Whitney test Wilcoxon test).
TaqMan Low Density miRNA Array (TLDA) Tiav
Peripheral blood tau sau los ntawm txhua tus neeg koom thiab muab xa mus rau qhov chaw kuaj ntshav hauv 4 h los txo cov ntshav cell lysis. Cov hnoos qeev tau centrifuged ntawm 3,000 rpm rau 10 min ntawm chav sov. Tom qab ntawd, supernatant (plasma txheej) tau sau tsis muaj kab mob hauv cov ntshav. Kev voj voog miRNAs tau muab rho tawm siv TaqMan miRNA ABC Purification Kit (Human Panel A; Thermo Fisher Scientific, Waltham, MA, USA) raws li qhov chaw qhia cov khoom qhia. Hauv kev qhia, 50 μL ntawm cov qauv ntshav thiab 100 μL ntawm ABC tsis tau sib xyaw. Tom qab kev cog qoob loo uas muaj kev sib tw tshwj xeeb los tiv thaiv me-miRNA sib sib zog nqus hlaws, bounded circulating miRNAs tau los ntawm cov hlaws dai nrog 100 μL ntawm elution tsis. Thaum 381 miRNAs raug soj ntsuam los ntawm 51 ntshav kuaj (25 IGDs thiab 26 controls) siv TaqMan miRNA ABC Purification Kit (Human Panel A, Thermo Fisher Scientific, Waltham, MA, USA) raws li cov khoom siv qhia. Megaplex thim rov qab thiab kev ua ntej cov lus thev ua ntej tau ua kom muaj qhov ntau ntawm cDNA rau kev miRNA qhia siv MegaplexPreAmp Primers Human Pool A thiab TaqManPreAmp Master Mix (Thermo Fisher Scientific). Lub vaj huam sib luag TLDA Lub v2.0 (Thermo Fisher Scientific) tau khiav ntawm ViiA7 real-time PCR system (Thermo Fisher Scientific) los ntsuas kev qhia txog cov miRNAs. Raw cov ntaub ntawv tau ua tiav kev siv ExpressionSuite Software v1.0.4 (Thermo Fisher Scientific) los txiav txim siab Ct rau txhua tus miRNA.
Kev Txheeb Xyuas Cov Ntsiab Lus rau TLDA
Peb ua ntej ntsuas ntsuas kev voj voog (Ct tus nqi) ntawm txhua miRNA. miRNAs uas muaj Ct tus nqi> 35 tau txiav txim siab tias yog qhov tsis paub thiab cais tawm ntawm kev tsom xam tom ntej. Txhua Ct qhov tseem ceeb tau ib txwm rau Ct tus nqi ntawm miR-374b (valueCt tus nqi), ib qho kev pom zoo tshaj plaws miRNA uas nthuav tawm hauv tib neeg ntshav (24). Log2 npawm-hloov piv (ΔΔCt tus nqi) ntawm qhov kev qhia tau muab xam siv qhov txiaj ntsim zoo ntawm kev kuaj kev kuaj ua tus calibrator hauv HTQPCR pob hauv Bioconductor (25). Qhov ntsuas kwv yees (RQ) ntawm txhua lub hom phiaj miRNA txhais tias 2-ΔΔCt. Rau kev ntsuas hypothetical ntawm qhov sib txawv hauv kev qhia ntawm ob pawg, peb thov surrogate kuj hloov (SVA) mus ntes heterogeneities xws li batch teebmeem hauv cov kev sim siv sva pob hauv Bioconductor (26). miRNAs nrog ib tug P-tus nqi <0.05 tau pom tias muaj ntau yam sib txawv ntawm ob pawg.
Noob Txhawb Kev Ntsuam Xyuas Kev Ntsuam Xyuas
Rau cov noob uas nrhiav kev pom zoo, peb siv ToppFun hauv ToppGene Suite (27) kom ua tau zoo dua qub Gene Ontology (GO) (28), cov ntsiab lus, txoj kab, thiab cov kab mob. Raws li cov tswv yim rau txoj kev qhia no, peb siv 1,230 los kwv yees cov noob ntawm tus neeg sib tw miRNAs. Kev ntsuam xyuas txoj hauv kev siv rau kev tshawb nrhiav tau siv los nrhiav cov kev tseem ceeb ntawm cov noob raws lub hom phiaj raws li KEGG, BioCarta, Reactome, GeneMAPP, thiab MSigDBin lub HompGene. Qhov tseem ceeb ntawm cov ntsiab lus zoo rau kev ua tau zoo raws li Bonferroni-kho P-peem.
Quantitative Reverse PCR (qRT-PCR) Kev Siv Tiag thiab Kev Lees
Yuav ua li cas thiaj paub tseeb tias 10 miRNAs uas tau sib txawv hauv qhov kev tshawb nrhiav pom, qRT-PCR tau siv cov TaqMan MicroRNA Assay (miR-15b-5p, # 000390; miR-26b-5p, # 000407; miR-29b-3p, # 000413; miR-125b-5p, # 000449; miR-200c-3p, # 002300; miR-337-5p, # 002156; miR-411-5p, # 001610; miR-423-5p, # 002340; miR-483 -5p, # 002338; thiab miR-652-3p, # 002352) thiab ViiA7 system (Lub neej ua haujlwm) raws li qhov kev tsim kho kev lag luam. Kaum nanograms ntawm tag nrho RNA tau hloov mus ua ntej-strand cDNA nrog cov qhia tseeb ntawm miRNA uas siv TaqMan MicroRNA Reverse Transcription Kit (#4366596, Lub neej Sojntsuam), raws li lub sijhawm PCR nrog TaqMan Probes. RQ ntawm txhua tus miRNA txhais tias 2-ΔCt, qhov twg ΔCt yog qhov txawv ntawm kev pib khoob mus rau qhov qauv nug, normalized tiv thaiv cov miRNA kawg (miR-374b-5p, # 001319). Tag nrho PCR cov kev xav tau raug coj tawm hauv kev sib txig, thiab lawv cov nqi Ct raug kwv yees. Peb xam lub log2 quav hloov (ΔΔCt) ntawm txhua tus miRNA tib txoj kev raws li hauv kev soj ntsuam raws array. Ib qho kev ntsuas tsis-parametric Mann-Whitney-Wilcoxon tau ntsuam xyuas qhov sib txawv hauv cov kev qhia ntawm miRNAs hauv ob pawg nrog ib qho chaw pib P-peem ntawm 0.05.
Western Blot Analysis
Txhua tus qauv ntshav yog thawj zaug ntawm cov tshuaj 14 high-abundance proteins (albumin, immunoglobulin G, immunoglobulin A, serotransferrin, haptoglobin, alpha-1 antitrypsin, fibrinogen, alpha-2 macroglobulin, alpha-1 acid glycoprotein, immunoglobulin M, apolipoprotein AI , apolipoprotein A-II, Nkaus C3, thiab transthyretin) uas siv cov kem MARS-14 (4.6 × 50 mm, Agilent Technology, Santa Clara, CA, USA) ua ntej hnub poob kev tshuaj ntsuam. Cov feem ntawm unbound uas tau los ntawm MARS-14 kem tau siv cov Amicon Ultracel Ultracel-3 centrifugal lim (3 kDa cutoff), thiab ces cov concentration protein tau txiav txim siab siv cov bicinchoninic acid method. Tus nqi tib yam (ntawm 10 rau 30 μg) ntawm kev tswj thiab kev kuaj ntshav IGD tau sib cais ntawm 4-20% Mini-PROTEAN TGX precast gel (Bio-Rad, CA, USA) thiab pauv mus rau ib lub hnab polyvinylidene difluoride membrane. Tom ntej no, lub membrane tau thaiv hauv TBS-T (190 mM NaCl, 25 mM Tris-HCl, pH XUM, thiab 7.5% Tween 0.05) nrog 20% tsis muaj rog mis nyuj haus hauv chav tsev kub rau 5 min. Cov tshuaj tua tau tom qab txhaj tshuaj tiv thaiv kab mob DPYSL30 (2, Novus Biologicals, Littleton, CO, USA), GABRB1 (500: 2, Abcam, Cambridge, MA, USA), thiab CNR1 (1000: 1, Santa Cruz Biotechnology (1: 100, MybioSource, San Diego, CA, Tebchaws USA) hauv TBS-T nrog 4, XYUAS, 1, 500, XYOOX, San Diego, CA, USA), thiab PI15 % tsis muaj roj mis nyuj rau ntawm 1 ° C thaum tsaus ntuj, thiab tom qab ntawd nrog rau cov tshuaj tiv thaiv kab mob tsim nyog (xws li 500, Santa Cruz Biotechnology) los yog tshis anti-luav (5: 4, Cell Signaling, Beverly, MA, USA ) conjugated rau horseradish peroxidase ntawm chav tsev kub rau 1 h. Qhov kev kuaj pom tshwm sim tau muab siv chemiluminescence nrog ECL reagent (GE healthcare, Piscataway, NJ, USA). Peb xam qhov nruab nrab ntawm cov txiaj ntsig thaj siv TotalLab 1,000D tsom xam software (Non-linear Dynamics, Newcastle upon Tyne, UK). Tom qab ntawd, tus nqi densitometry piv tau los ntawm kev faib cov densitometry tus nqi ntawm txhua tus qauv li tau piav nyob rau lwm qhov (29). Raws li kev tswj rau normalization, ib qho kev kuaj ntshav tau los ntawm 46 IGD thiab tswj kev kuaj tau siv rau txhua txoj kev sim. Qhov tseem ceeb ntawm kev txiav txim tau raug txiav txim siab siv ib qho kev xeem tsis tshua meej ntawm Mann-Whitney-Wilcoxon nrog ib qho chaw pib P-peem ntawm 0.05.
tau
Yam ntxwv ntawm Cov Kawm Tshawb Fawb
Cov xwm txheej ntawm pejxeem thiab soj ntsuam ntawm cov kev kawm tseem muaj qhia nyob rau hauv Table Table1.1. Thaum peb piv IGD thiab tswj hwm pawg raws li Kauslim Teb Internet Kev Npliag Kev Ntsuas (K-Scale) raws li piav hauv lwm qhov chaw (20, 30), pab pawg neeg IGD pom ib tus nqi siab dua K-nplai tshaj qhov kev tswj hwm (37 vs. 24, P = 3.81 × 10-6) (Cov lus (Table1). 1). Cov neeg Median txhua lub lim tiam siv sijhawm hauv Internet gaming hauv pab pawg neeg IGD tau ntev dua li ntawm cov tswj (18 vs. 5.25 h, P = 1.27 × 10-6). Whereas tsis muaj qhov sib txawv ntawm ob pawg hauv lub hnub nyoog, cov nyiaj tau los hauv tsev neeg, kev kawm ntawv, kev tsim qauv, thiab cov ntsiab lus ntawm K-WISC, BIS, BInS, thiab BAS.
Txawv Txawj Hais Tiag txog miRNAs Ntawm IGD thiab Ntawm Cov Kev Tswj
Yuav nrhiav tau IGD-miRNAs, peb saws ob-kauj ruam (discovery thiab ywj siab validation). Txoj kev tshawb no thiab kev tawm tswv yim zoo yog qhia hauv Daim Ntawv S1 hauv Cov Ntawv Qhia Ntxiv. Nyob rau hauv lub discovery theem, peb analyzed miRNA qhia profiles ntawm 51 kuaj (25 IGDs thiab 26 ntawm lwm yam uas) siv cov miRNA array uas muaj 384 miRNAs. Qhov qhia ntawm 10 miRNAs tau pom tias nws txawv ntawm IGD thiab pawg tswj hwm (Cov Lus (Table2). 2). Cov txheeb ze qhia qib ntawm cov 10 miRNAs yog muaj nyob rau hauv Xam Xam Figure1.1. Ntawm lawv, ob tug (hsa-miR-423-5p thiab hsa-miR-483-5p) tau muab kho thiab yim (hsa-miR-15b-5p, hsa-miR-26b-5p, hsa-miR-29b-3p, hsa-miR-125b-5p, hsa-miR-200c-3p, hsa-miR-337c-5p, hsa-miR-411-5p, thiab hsa-miR-652-3p) raug txo nyob hauv pawg IGD.
rooj 2
Qhov sib txawv ntawm qhov sib txawv ntawm cov microRNAs (miRNAs) thiab cov kev hloov quav.
| miRNA | Discovery | Validation | Ua ke | |||
|---|---|---|---|---|---|---|
| P-peem | Quav hloov | P-peem | Quav hloov | P-peem | Quav hloov | |
| hsa-miR-15b-5p | 0.033 | 0.829 | 0.694 | 1.119 | 0.381 | 0.947 |
| hsa-miR-26b-5pa | 0.008 | 0.871 | 0.049 | 0.841 | 0.013 | 0.857 |
| hsa-miR-29b-3p | 0.005 | 0.400 | 0.560 | 1.187 | 0.089 | 0.647 |
| hsa-miR-125b-5p | 0.021 | 0.582 | 0.290 | 0.950 | 0.069 | 0.723 |
| hsa-miR-200c-3pa | 0.011 | 0.336 | 0.003 | 0.542 | 2.93 × 10-5 | 0.415 |
| hsa-miR-337c-5p | 0.009 | 0.385 | 0.582 | 0.872 | 0.020 | 0.553 |
| hsa-miR-411-5p | 0.004 | 0.322 | 0.336 | 1.282 | 0.158 | 0.595 |
| hsa-miR-423-5p | 0.026 | 1.387 | 0.189 | 0.955 | 0.518 | 1.175 |
| hsa-miR-483-5p | 0.018 | 1.861 | 0.765 | 1.413 | 0.211 | 1.647 |
| hsa-miR-652-3pa | 0.019 | 0.715 | 0.049 | 0.877 | 0.011 | 0.782 |
amiRNAs tau hloov kho hauv ob qho tib si kev tshawb nrhiav thiab kev lees paub tseeb hauv kev sib luag.
qRT-PCR Validation ntawm tus neeg sib tw miRNAs
Yuav kom paub tseeb tias 10 tus neeg sib tw miRNAs, peb tau ua qRT-PCR nrog ib qho kev tso cai ywj pheej (20 IGDs thiab 16 controls) (Cov lus S1 nyob rau hauv Supplementary Materials). Peb ntawm cov miRNAs (hsa-miR-200c-3p, hsa-miR-26b-5p, thiab hsa-miR-652-3p) tau txo qhov teeb meem hauv IGD pawg ntawm qhov kev lees paub (Table2). 2). Peb lwm cov miRNAs (hsa-miR-337c-5p, hsa-miR-125b, thiab hsa-miR-423-5p) kuj tau downregulated nyob hauv pab pawg IGD tab sis tsis tseem ceeb. Tshuav plaub miRNAs (hsa-miR-15b-5p, hsa-miR-29b-3p, hsa-miR-411-5p, thiab hsa-miR-423-5p) tau hais tawm tsam qhov teeb tsa kev cai. Thaum peb ua ke cov kev tshawb nrhiav thiab kev lees paub (tag nrho ntawm 45 IGD kawm thiab 42 tswj), peb cov validated miRNAs tau xwm yeem tseem ceeb (Cov lus (Table2). 2). Cov ncauj lus kom ntxaws, qhov chaw chromosomal, cov zauv tshwm sim, thiab qhia ntau ntau hauv CNS ntawm no cov miRNAs peb muaj nyob rau hauv Table S2 hauv Cov Ntawv Qhia Ntxiv.
Synergistic nyhuv ntawm Simultaneous Hloov ntawm peb cov miRNAs ntawm IGD yuav raug
Kev ntsuam xyuas qhov kev txuam ua ke ntawm peb cov miRNAs, peb pom qhov sib txawv ntawm qhov sib txawv (ORs) ntawm plaub subgroups (nrog 0, 1, 2, lossis 3 miRNA alterations). miRNA hloov tau txhais los ntawm tus nqi RQ li tau hais tseg hauv Tshooj lus "Ntaub ntawv thiab kev"Vim hais tias tag nrho peb cov miRNAs cov cim tau downregulated nyob rau hauv pab pawg neeg IGD, tus miRNA uas nws tus nqi RQ nyob hauv qab yog ib qho defied li hloov kho ib qho. Muaj cov ncauj lus kom ntxaws txog txhua qhov kev kawm hauv qhov kev kawm RQ rau peb cov miRNAs muaj nyob hauv Cov Ntawv S3 hauv Cov Ntawv Qhia Ntxiv. Rau txhua pawg ua ke, qhov sib npaug raug muab xam tias qhov sib npaug ntawm cov kev tswj ntawm cov IGDs, ces txhua tus LOSSIS tau muab xam los ntawm qhov faib txhua feem ntawm txhua pawg uas muaj pes tsawg tus tsis muaj kev hloov ntawm miRNA. Cov neeg uas muaj peb qho kev ntsuas meRNA pom tau hais tias yuav muaj 22 lub sijhawm siab tshaj qhov tsis muaj kev hloov ntawm miRNA (LOS YOG 22, 95% CI 2.29-211.11). ORs tau pom ib qho kev nce zuj zus nrog cov kev hloov kho miRNAs ntawm 0 rau 3 (r2 = 0.996) (Daim duab (Figure22).
GO thiab Pathway Analysis ntawm Hom Phiaj Ntawm Cov Neeg Sib Tw miRNAs
Mus nrhiav kev pom rau hauv cov dej num ntawm peb lub miRNA cov cim ntsuas ntawm cov IGD hauv pawg IGG, lawv lub hom phiaj tau kwv yees siv cov miRWalk 2.0 database (31). Ib qho tag nrho ntawm 1,230 lub noob tau xwm yeem tau muab pov tseg raws li kev nqes taw ntawm plaub lub ntsiab ntawm plaub algorithms (miRWalk, miRanda, RNA22, thiab Targetscan) siv cov miRWalk database (32-34) (Cov lus S4 nyob rau hauv Supplementary Materials). Noob muaj kev paub ntau ntxiv uas siv ToppFun hauv ToppGene Suite pom tias cov hom phiaj ntawm cov miRNAs tau txuam nrog txoj kev loj hlob neural xws li "Axon kev taw qhia" thiab cov nqe lus xws li "neurogenesis" (Cov lus S5 nyob rau hauv Supplementary Materials).
Qhia tawm ntawm Cov Hom Phiaj Ntiag Tug
Ntawm cov phiajcim qis ntawm peb lub miRNAs, 140 tau kwv yees li ob lossis ntau dua miRNAs (Cov lus S4 nyob rau hauv Supplementary Materials). Yog xav paub seb lawv cov kab lus protein ntau npaum li cas ntawm cov nqes dej hauv plab yog txawv ntawm IGD thiab pawg tswj hwm, peb xaiv 2 noob (DUSP4 thiab PI15), uas yog kwv yees raws kev nqes taw ntawm tag nrho cov 3 miRNAs thiab ntxiv 3 noob (GABRB2, DPYSL2, Thiab CNR1) los ntawm cov uas tau hais tseg rau 2 miRNAs thiab tau siv thaj poob tsom nrog cov ntshav kuaj los ntawm 28 IGDs thiab 28 ntawm cov khoom siv rau kev sim. Peb piv cov lus ntawm tsib lub hom phiaj ntawm IGD thiab pawg tswj hwm los ntawm kev ntsuas cov qhab nia thiab cheeb tsam raws li tau piav nyob rau lwm qhov (29). Ntawm lawv, qhia qib DPYSL2 (28 IGDs thiab 28 tswj, P = 0.0037) thiab GABBR2 (27 IGDs thiab 28 tswj, P = 0.0052) tau nce siab hauv IGD pawg (Daim duab (Figure3). 3). Txawm li cas los xij, peb tsis pom cov lus sib txawv ntawm CNR1 (P = 0.0853), DUSP4 (P = 0.5443), thiab PI15 (P = 0.6346).
kev sib tham
Nws tau raug qhia tias cov miRNAs tau koom tes rau hauv kev loj hlob neuronal (35, 36), thiab qhia tawm ntawm lub hlwb hlwb miRNAs raug pom hauv cov kev mob hlwb xws li schizophrenia (37). Yog li, nws yog plausible tias circulating miRNA profiles yuav pab tau biomarkers rau IGD. Kev voj voog miRNAs tau pom tias yog biomarkers rau ntau hom kev mob hlwb neuropsychiatric (38-40); Txawm li cas los xij, cov txheej txheem molecular mechanisms tom qab IGD txoj kev loj hlob yog tseem tsis tau paub ntau dua li nws txoj kev kuaj mob thiab tseem ceeb. Tshwj xeeb, tsis muaj kev tshawb fawb txog IGD-miRNAs. Lub hom phiaj ntawm txoj kev tshawb fawb no yog ob qho tib si. Ua ntej, peb tau sim mus nrhiav ntshav cov miRNAs nrog IGD. Qhov thib ob, peb soj ntsuam qhov txheeb raws roj ntsha ntawm miRNA los ntawm kev tshawb xyuas cov protein thiab GO ntawm cov hauv paus noob. Los ntawm kev tshawb nrhiav genome-wide ntawm miRNA qhia txog kev sib raug zoo thiab kev nqes peev ntawm qhov kev sib tw ntawm qhov kev sib tw, peb pom tias kev qhia peb cov miRNAs (hsa-miR-200c-3p, hsa-miR-26b-5p, thiab hsa-miR-652-3p) ho qis dua hauv cov neeg mob IGD dua ntawm cov tshuaj. Txawm hais tias cov qauv ntawm lwm tus xya miRNA sib tw tsis tau muab rov los ua ke, nws tuaj yeem yog cov lus tsis zoo vim tias tus qauv me me hauv qhov kev tshawb fawb no. Rau peb txoj kev paub, qhov no yog thawj daim ntawv qhia txog qhov tseem tias ntshav miRNA qhia profiles yuav pab tau biomarkers rau IGD. Kev sib xyaw ntawm peb lub cim ntawm miRNA tuaj yeem ua ib qho cuab yeej tsawg heev rau cov neeg thaum ntxov ntawm cov neeg pheej hmoo ntawm IGD.
Cov miRNA uas tau txheeb xyuas hauv txoj kev tshawb no tau raug tshaj tawm tias muaj kev koom tes rau ntau hom kev puas hlwb neuropsychiatric. Kev qhia ntawm hsa-miR-200c hauv cov ntshav tau raug tshaj tawm hauv cov kev puas siab puas ntsws xws li schizophrenia (41) thiab kev nyuaj siab loj (42). miR-200c tau tshaj tawm tias muaj ntau tshaj tawm hauv cov kev sib txuam ua ke tshaj li hauv tag nrho forbain (43) thiab kuj yog txuam nrog kev tuag ntawm kev tuag ntawm neuronal (44). Raws li cov lus qhia dhau los no, miR-200c tau koom tes rau hauv kev tsim kho mob neurodevelopment thiab tuaj yeem txuam nrog mob paj hlwb (neuropsychiatric disorders) yog tias nws qhov kev qhia tsis zoo. Muaj ntau cov kev tshawb fawb pom tias muaj kev sib koom tes ntawm miR-652 thiab muaj kev puas siab ntsws mob hlwb. Zoo ib yam li peb txoj kev qhia, kom paub cov ntshav biomarkers rau schizophrenia, Lai et al. nqa tawm TLDA cov kev ntsuam xyuas nrog cov neeg mob schizophrenia thiab cov kev tswj mob, thiab pom tias xya miRNAs nrog hsa-miR-652 tau sib txawv ntawm cov neeg mob schizophrenia (45). Nyob rau hauv txoj kev tshawb tom ntej, lawv tsim ib qho kev twv ua ntej uas yog siv cov ntaub ntawv qhia miRNA thiab ntse ntse txawv schizophrenia los ntawm kev tswj hwm (46). Hloov kev hloov ntawm hsa-miR-652 kuj tau pom nyob hauv alcoholics (47). Hsa-miR-26b tau pom tias yuav tsum tau qhib lub zog thaum lub sijhawm sib txawv ntawm neuronal (48). Perkins li al. qhia hais tias hsa-miR-26b tau nyob rau hauv cov hauv cov neeg mob cov paj hlwb (prefrontal cortex) ntawm tus neeg mob schizophrenia (49).
Txawm hais tias tsis muaj cov pov thawj ncaj nraim los txhawb txoj kev sib raug zoo ntawm cov perturbed qhia ntawm cov miRNAs thiab cov pathophysiology ntawm IGD, peb tuaj yeem xav tias qhov dysregulation ntawm cov miRNAs yuav txuam nrog cov pathophysiology ntawm IGD raws li ntau cov lus qhia dhau los ntawm cov qis nqus peb kwv yees . Qee cov nram qab rog ntawm peb cov miRNAs xws li GABRB2, CNR1, NRXN1, Thiab DPYSL2 raug tshaj tawm qhia tias yog mob dab tsi nrog rau kev puas hlwb neuropsychiatric disorders. Gamma-aminobutyric acid (GABA) yog ib qho tseem ceeb inhibitory neurotransmitter hauv CNS. Dysregulation ntawm GABA tus neeg tau txais kev pabcuam, yog muaj kev cuam tshuam rau kev mob hlwb (neuropsychiatric disorders) nrog kev tivthaiv, kev ntxhov siab, thiab kev nyuaj siab (50), uas yog cov tseem ceeb ntawm IGD (8). Genetic polymorphisms nyob rau hauv GABA receptor noob yog qhia txog kev haus dej cawv thiab tus mob schizophrenia (51, 52). Dihydropyrimidinase zoo li 2 (DPYSL2) yog ib tug tswvcuab ntawm cov neeg sib tw rau cov neeg sib tw rau tus neeg pluag, uas yog lub luag haujlwm hauv cov kev sib txuas lus ntawm microtubule, kev sim tshuaj, thiab kev tswj kev loj hlob ntawm txoj kev loj hlob. Txawm li ntawd los, qhov no tau muab tso ua ke raws li biomarker rau kev puas siab puas ntsws (53, 54). Polymorphism hauv lub DPYSL2 noob kuj tau raug tshaj tawm los mus nrog cawv siv tsis meej (55). Cov ntaub ntawv yav dhau los thiab peb cov ntaub ntawv hais tias overexpression ntawm GABRB2 thiab DPYSL2, cov phiaj xwm kev nqes ntawm cov miRNAs downstulated, muaj qhov cuam tshuam rau cov pathogenesis ntawm neuropsychiatric disorders, xws li IGD. Cannabinoid receptor type 1 (CNR1) yog ib hom tshuaj haj yam tshuaj (presidaptic heteroreceptor) uas kho cov neurotransmitter tso tawm thiab cov kab mob hauv cannabinoid signaling muaj ntau yam kab mob neuropsychiatric disorders (56). Genetic polymorphism ntawm CNR1 Noob ntsej muag yog paub txog kev quav dej quav cawv nyob rau hauv Caucasians (57). Hauv kev ua qauv zoo, kev ua kom ventral hippocampus CNR1 cuam tshuam kev coj tus cwj pwm zoo thiab cognition (58). Kev hloov caj ces nyob hauv NRXN tsev neeg yog paub tias muaj kev koom tes rau ntau hom kev mob paj hlwb (neuropsychiatric disorders) nrog rau kev quav tshuaj (59).
Txheeb xyuas qhov kev txheeb raws roj ntsha ntawm peb cov miRNA sib tw nyob rau hauv ib txoj kev ncaj ntxiv, peb tau tshawb txog cov khoom protein tawm ntawm lawv cov noob qis. Vim tias muaj tsawg ntawm cov kev kuaj ntshav, ntawm 140 cov neeg sib tw (kwv yees raws li nram 2 los yog ntau miRNAs), peb tau soj ntsuam 5 lub hom phaj (GABRB2, DPYSL2, CNR1, DUSP4, thiab PI15) los ntawm thaj poob thiab lees tias qhov qhia ntawm GABRB2 thiab DPYSL2 tau yooj yim dua hauv pawg IGD. Cov ntaub ntawv qhia yav dhau los thiab peb cov ntaub ntawv hais tias overexpression ntawm GABRB2 thiab DPYSL2, cov phiaj xwm kev nqes ntawm cov miRNAs downstulated, yuav muaj qhov cuam tshuam rau cov pathogenesis ntawm neuropsychiatric disorders, xws li IGD. Cov qhab-nees ntawm GO thiab pathway tsom neural development pathways kuj pab txhawb lub hlwb kev xav ntawm cov miRNA. Lwm cov ntsiab lus nrhiav tau yog qhov kev txuam ntawm synultistic ntawm kev hloov ntawm cov miRNAs. Cov neeg uas raug txo ntawm tag nrho cov 3 miRNAs pom 22 lub sijhawm ntau dua li cov uas tsis muaj kev txo qis qis, thiab cov ORS tau nce siab ntau dua. Txawm hais tias CI rau cov kev hloov no peb tau dav vim qhov loj me me, qhov pom tseeb zoo sib xws (r2 = 0.996) txhawb nqa qhov ua haujlwm synergistic ntawm peb miRNA.
Txawm hais tias peb tau pom IGR-kis miRNA cov cim thiab cov tib neeg nrog tag nrho peb cov miRNA alteration muaj 22 lub sij hawm siab tshaj cov tsis muaj kev miRNA alterations, muaj ntau ntau cov kev txwv hauv txoj kev tshawb no. Ua ntej, qhov ua piv txwv me me nce ntxiv yuav ua rau muaj lwm qhov cim npe miRNA. Thib ob, txij li thaum peb cov ntaub ntawv tsis txaus los qhia kom meej tias cov ntshav miRNA profiles yog leej twg ua rau los sis cuam tshuam, peb tsis tuaj yeem tshawb xyuas cov haujlwm ntawm caj dab tsis ntawm cov chaw kho mob. Ntxiv miRNA profiling thiab lawv cov kev nqes roj hauv qab siv cov tib neeg lub hlwb cov nqaij los ntawm paj hlwb nqaij muaj peev xwm muab tau lus teb ntxiv rau. Lub paj hlwb cov tsom xam nrog cov kev ua si tsis zoo rau tus tsiaj kuj yuav pab tau. Thib peb, vim muaj kev txwv cov ntshav qis xwb, peb tsuas kuaj tsib tus neeg sib tw ntawm cov neeg sib tw. Tshawb nrhiav ntau lub phiaj xwm nrog cov qauv loj dua yuav pab tau kom nkag siab ntxiv txog txoj kev molecular mechanism ntawm IGD.
Hauv cov ntsiab lus, los ntawm kev soj ntsuam genome thoob plaws ntawm miRNA qhia cov ntaub ntawv thiab kev ywj siab, peb nrhiav tau peb IGD-txuas miRNAs (hsa-miR-200c-3p, hsa-miR-26b-5p, thiab hsa-miR-652-3p). Ntau yam hauv lawv cov qog nqus tau raug tshaj tawm hais tias muaj kev koom tes rau ntau hom kab mob hlwb, thiab qhov kev hloov ntawm kev hloov ntawm cov caj qaum hauv qab no txhawb qhov kev siv ntawm miRNAs txheeb xyuas qhov kev tshawb fawb no. Peb pom tias cov neeg uas muaj kev txo ntawm tag nrho peb cov miRNA yog ntxim yuav raug tus kab mob IGD. Ua ke nrog cov kev kuaj mob los yog ib puag ncig muaj kev puas tsuaj thiab kev ntsuam xyuas pom, peb cov kev tshawb nrhiav tuaj yeem pab txhawb cov kev pabcuam thaum ntxov los pab rau cov neeg muaj feem raug mob siab tshaj ntawm IGD.
Nqe Kev Ncaj Ncees
Txoj kev tshawb no tau pom zoo los ntawm Institutional Review Board ntawm Catholic University Kawm Kev Kawm Txuj Ci Qib Kauslim (MC16SISI0120). Tag nrho cov neeg tuaj koom thiab lawv niam lawv txiv tau sau daim ntawv pom zoo.
Sau cov tswv yim
ML thiab HC tau sib koom nrog rau daim ntawv no. ML, D-JK, thiab Y-JC tsim qhov kev tshawb fawb. SJ, S-MC, YP, DC, thiab JL ua cov kev sim thiab cov ntaub ntawv qhia. J-WC, S-HP, J-SC, thiab D-JK sau cov ntshav kuaj thiab cov ntaub ntawv kuaj mob. ML, HC, S-HY, thiab Y-JC soj ntsuam cov ntaub ntawv. ML, HC, S-HY, thiab Y-JC tau piav txog kev sau ntawv. Y-JC tau saib xyuas txoj haujlwm.
Teebmeem ntawm Cov Lus Qhia Txaus
Cov sau phau ntawv tshaj tawm tias qhov kev tshawb fawb tau ua thaum tsis muaj kev lag luam los yog nyiaj txiag kev sib raug zoo uas yuav raug txwv tsis pub muaj kev cuam tshuam ntawm kev txaus siab.
Tshooj ntawv
Nyiaj. Cov haujlwm no tau txais kev pabcuam los ntawm Kev Pabcuam Kev Txawj Ntse los ntawm National Research Foundation ntawm Kauslim Teb (NRF), nyiaj txiag los ntawm Ministry of Science thiab ICT thiab Yav Tom Ntej Planning (NRF-2015M3C7A1064778).
Cov Ntaub Ntawv Ntxiv
Cov ntaub ntawv ntxiv rau cov ntsiab lus no muaj nyob hauv online ntawm http://www.frontiersin.org/articles/10.3389/fpsyt.2018.00081/full#supplementary-material.
References
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