Chapter1: Peroxisomes tubulaires ren yo disponib pou fonksyon ren nòmal
Jun 10, 2022
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Peroxisomes yo se òganèl selilè espesyalize ki enplike nan yon varyete pwosesis metabolik. Nan imen, mitasyon ki mennen nan pèt konplè nan peroxisomes lakòz echèk miltiògàn (maladi spectre Zellweger a, ZSD), ki gen ladanensifizans ren. Sepandan, (pato)fizyolojik wòl peroxisomes nanrenrete enkoni. Nou te adrese wòl peroxisomes nanfonksyon rennan sourit ak ablation kondisyonèl nan byogenesis peroxisomal nan tubul ren (cKO sourit). Analiz fonksyonèl pa t 'revele okenn fenotip ren ouvè nan sourit cKO. Sepandan, ti bebe cKO sourit gason te gen pi ba kò ak pwa ren, ak granmoun sourit gason cKO te montre rediksyon sibstansyèl nan pwa ren ak pwa ren/pwa kò. Analiz estereolojik te montre yon ogmantasyon nan dansite mitokondri nan selil tubul proximal sourit cKO. Analiz transcriptome ak metabolom entegre te revele repwogram pwofon plizyè wout metabolik, ki gen ladan metabolis glutatyon ak byosentèz / biotransformation nan plizyè gwo klas lipid. Malgre ke analiz sa a sijere konpanseestrès oksidatif, defi a ak manje ki gen anpil grès pa t 'pwovoke defisyans ren enpòtan nan sourit cKO. Nou demontre ke peroxisomes tubulaires ren yo dispansab pou fonksyon ren nòmal. Done nou yo tou sijere ke andikap ren nan pasyan ki gen ZSD yo gen orijin ekstrarenal.
Klike la a pou aprann sou benefis ekstrè cistanche tubulosa
Entwodiksyon
Peroxisomes se yon sèl-manbràn òganèl ki mare ke yo te jwenn premye nan ren sourit pa Rhodin an 1954(1). Estimasyon aktyèl sijere ke peroxisomes gen plis pase 50 anzim ki enplike nan yon varyete fonksyon selilè, ki gen ladan -oksidasyon nan asid gra chèn trè long (VLCFAs), asid gra chèn long (LCFAs), ak asid dicarboxylic long chèn; a-ak-oksidasyon nan asid gra chèn branch; oksidasyon nan prostaglandin ak leukotrienes; metabolis nan asid amine; byosentèz etè fosfolipid (ki gen ladan plasmalogens) ak asid bile; omeyostazi redox; dezentoksikasyon glyoxylate; ak ferroptoz. Peroxisomes yo prezan omniprésente nan prèske tout selil mamifè yo, ak abondans ki pi wo nan selil tubul proximal ren ak epatosit. Nan selil sa yo, peroxisomes okipe apeprè 3 pousan nan volim selil yo, men kantite yo, gwosè ak fòm yo varye anpil pandan devlopman anbriyon ak apre anbriyon (2); kantite yo ka grandi anpil nan divès kondisyon estrès (3). Malgre ke anpil anzim peroxisomal yo te byen karakterize, syans adrese ekspresyon yo ak wòl fonksyonèl nan ren yo ra. Anplis, enpòtans fonksyonèl an jeneral nan peroxisomes nan ren an rete lajman enkoni.
Prèv pou wòl peroxisomes nan ren (patho)fizyoloji te sitou parèt nan etid jenetik imen. Yo te idantifye de kalite mitasyon ki mennen nan maladi peroxisomal imen: (i) mitasyon nan sa yo rele peroxin (PEX) jèn ki enplike nan byogenesis peroxisome ak (ii) mitasyon nan anzim peroksizomal espesifik. Premye kalite mitasyon an mennen nan malfonksyònman peroksizomal jeneralize ki lakòz maladi espèk Zellweger (ZSD) (4). Tibebe ki gen fòm ZSD grav anjeneral mouri pandan premye ane a nan lavi akòz echèk milti-òganik. Menmsi prezans ren kortikal ak/oswa wòch oksalat ren nan pasyan ki gen ZSD te byen dokimante pandan plizyè ane (5, 6), mekanis molekilè ki mennen nan andikap sa yo rete byen lwen tèlman enkoni. Lòt anomali ren posib nan ZSD pa te envestige paske maladi a domine pa konplikasyon newolojik. Enpak fòm entèmedyè oswa pi modere ZSD sou fonksyon ren pa te evalye sistemikman. Prèv ki lye yon sèl defisi anzim peroksizomal ak fizyopatoloji ren tou rete limite.
Mitasyon nan alanin peroxisomal fwa a: glyoxylate aminotransferase kode pa jèn AGXT mennen nan iperoksaluri prensipal kalite I, yon maladi karakterize pa depo kristal oksalat kalsyòm nan ren an (revize nan ref. 7). Dènyèman, yo te idantifye yon mitasyon otozomal dominan ki mennen nan sendwòm Fanconi nan ren, oswa malfonksyònman jeneralize nan tib proximal la, nan anzim peroxisomal enoyl-CoA idrataz ak 3-hydroxy acyl CoA dehydrogenase (EHHADH)(8) . Mitasyon sa a lakòz mistargeting nan EHHADH nan mitokondri ak andikap fonksyon mitokondriyo. Etid jenetik ki fèt sou tansyon rat konjenik yo te montre ke anzim peroxisomal idwoksi asid oksidaz 2(HAO2) ka jwe yon wòl nan kontwòl tansyon (9,10). Sepandan, si fenotip sa a se akòz fonksyon HAO2 ki chanje nan ren an oswa lòt tisi rete enkoni. An jeneral, done ki soti nan modèl bèt ki gen defo espesifik nan ren nan byogenesis peroxisome oswa inaktivasyon espesifik nan ren nan yon sèl anzim peroxisomal yo toujou manke. Isit la, nou adrese wòl peroxisomes nan fonksyon ren nan sourit gason ak fi ak ablasyon kondisyonèl nan byogenesis peroxisomal nan tib ren.
Rezilta yo
Jenerasyon ak karakterizasyon debaz sourit gason ak fi tibebe ak ablation kondisyonèl nan byogenesis peroxisomal nan tubul ren an. Peroxisome byogenesis nan tib ren yo te deranje nan inaktivasyon kondisyonèl nan Pex5 kode cytosolic peroxisome vize siyal 1 (PTS1) reseptè, ki se esansyèl pou enpòte nan PTS1 motif ki gen pwoteyin nan peroxisomes (11,12). Suppression kondisyonèl nan Pex5 nan tubul ren yo te reyalize lè l sèvi avèk yon sistèm doxycycline-inducible (DOX-inducible) (PexSanlo/Pax8-rtTA/LC1 sourit, ref.13). Paske enpòtans fonksyonèl peroxisomes yo ka depann de laj (14), yo te fè karakterizasyon debaz defisyans Pex5 nan ren nan tou de sourit tibebe ak adilt. Nan eksperyans ak sourit tibebe yo, yo te fè eksizyon nan alèl Pex5 floxed atravè administrasyon DOX (2 mg/mL nan dlo pou bwè) nan fi ansent nan E14 epi kenbe jiskaske P7. Sourit tibebe yo te sakrifye nan P28. Sourit ak jenotip Pex5oilo yo te itilize kòm kontwòl. Jan yo montre nan Figi Siplemantè 1A (materyèl siplemantè ki disponib sou entènèt ak atik sa a; https://doi.org/10.1172/jci.insight.155836DS1), tretman DOX te lakòz yon eksizyon prèske konplè nan alèl Pex5 floxed nan Pexswn/Pax{{ 29}}GTA/LC1 sourit ti bebe gason ak fi. Analiz de seksyon ren yo pa t 'revele okenn anomali istolojik aklè oswa wòch ren nan sourit tibebe san Pex.5 nan tubul ren (pa montre). Albuminuria te absan nan echantiyon pipi tach sourit Pex5-devoid ti bebe nan tou desèks(Figi Siplemantè 1B). Pwa kò (BW) ak pwa ren (KW) men pa rapò KW/BW yo te pi ba nan sourit gason Pex 5-devoid tibebe konpare ak kontwòl littermate (Figi Siplemantè 1C).
Jenerasyon ak karakterizasyon debaz sourit granmoun gason ak fi ak ablasyon kondisyonèl nan byogenesis peroxisomal nan tubul ren an. Sipresyon Pex5 nan tubul ren sourit adilt yo te pwovoke pa tretman 2-semèn ak DOX nan 8-semèn Pex5xio/Pax8-ntTA/LC1 sourit gason ak fi, yo refere yo aprè sa a. kòm sourit cKOm ak cKOf, respektivman (gade Metòd) .An paralèl, yo te bay menm tretman DOX nan kontwòl littermate yo (sourit Pex5a/lx, yo rele sourit Ctrlm ak Ctrlf, respektivman). Tout eksperyans sou sourit adilt yo te fèt 4 semèn apre fen tretman DOX. Jan yo montre nan Figi 1A, ekspresyon Pex5 mRNA te siyifikativman redwi nan ren tou de sourit cKOm ak cKOf. Sepandan, nan sourit cKOf, kèk rezidyèl plen longè Pex5 mRNA te detekte tou, sijere ke eksizyon enkonplè nan alèl floxed la. Menm jan an tou, pwoteyin PEX5 te nòmalman absan nan ren yo nan sourit cKOm men yo toujou detekte nan sourit cKOf, byenke nan yon nivo sibstansyèlman pi ba. Diferans sa a te vizib lè ekstrè ren ki soti nan cKOm ak cKOf sourit yo te chaje sou menm jèl SDS-PAGE la ak imunoblotted ansanm (Figi 1B). Tou de sourit Kim ak cKOf yo te solid, pa montre okenn anomali aklè, epi yo te gen BW nòmal konpare ak sourit kontwòl (Tablo Siplemantè 1). Rapò KW ak KW/BW yo te pi ba anpil nan sourit cKOm konpare ak sourit Ctrl men se pa nan sourit cKOf konpare ak sourit Ctrlf (Figi 1, C, ak D, respektivman). Analiz 24-èdtan echantiyon pipi ak plasma pa t 'revele yon efè jenotip nan sourit tou de sèks, eksepte pou pi ba nivo potasyòm plasma nan sourit cKOm konpare ak sourit Ctrl (Tablo Siplemantè 1). Albuminuria te absan nan pipi tou de sourit cKOm ak cKOf (figi siplemantè 2A). Pa gen okenn chanjman mòfolojik brit, depo oksalat kalsyòm, oswa akimilasyon lipid yo te obsève nan ren sourit cKOm (Figi Siplemantè 2, BD, respektivman).
Evalyasyon mikwoskopi elektwonik nan selil tubul proximal nan sourit adilt cKO. Evalyasyon mikwoskospi elektwonik nan cortical ren an te revele yon gwo kantite peroxisomes nan selil tubul proximal sourit Ctrlm ak Ctrlf (Figi 2, A, ak B, respektivman). Peroxisomes te nòmalman absan nan tubul proximal nan sourit cKOm ak cKOf (Figi 2, Cand D, respektivman). Zouti estereolojik yo te anplwaye pou analiz quantitative nan selilè
òganèl ak dimansyon selilè nan tubul proximal sourit Ctrlm ak cKOm. Tou de dansite volim peroxisomes '(nimewo / μm² sitoplasm) ak pousantaj nan sitoplasm okipe pa peroxisomes nan selil tubul proximal yo te dramatikman diminye nan sourit cKOm (Figi 3, A ak B, respektivman), Kontrèman, mitokondriyo volim dansite osi byen, pousantaj nan sitoplasm okipe pa mitokondri nan selil tubul proximal, te ogmante nan sourit cKOm konpare ak sourit Ctrl (Figi 3, C ak D, respektivman). Epitou-vini dansite volim ak pousantaj nan sitoplasm okipe pa lizozom nan selil tubul proximal yo pa t diferan ant sourit Kim ak Ctrlm (Figi 3, E ak F, respektivman). Jan yo montre nan Figi 3G, selil tubul proximal ki soti nan sourit cKOm yo te gen tandans redwi lajè selil (P=0.083).
Analiz transcriptomic ak metabolomik entegre sijere repwogram pwofon nan metabolik, antioksidan, ak wout sentèz lipid nan ren yo nan sourit cKO. Entegre transcriptome-sekans ak analiz metabolom yo te fèt pou idantifye chanjman molekilè nan ren sourit cKO (GSE179202). Konparezon transkriptòm yo te revele 1350 transkripsyon diferan ki eksprime nan ren sourit Ctrlm ak cKOm (Figi 4A ak Tablo Siplemantè 2, FDR).< 5%)and="" 121="" transcripts="" differentially="" expressed="" in="" kidneys="" of="" ctrlf="" and="" ckof="" mice="" (figure="" 4b="" and="" supplemental="" table="" 3,="">< 5%).="" 61="" differentially="" expressed="" transcripts="" were="" present="" in="" both="" sexes(figure="" 4c="" and="" supplemental="" figure="" 3).="" enrichment="" analysis="" of="" 100="" genes="" encoding="" proteins="" related="" to="" peroxisomal="" function(kyoto="" encyclopedia="" of="" genes="" and="" genomes="" [kegg]="" pathway="" hsa04146)performed="" on="" transcriptomes="" of="" ctrlm="" and="" ckom="" mice="" revealed="" 52="" transcripts="" either="" up-or="" downregulated="" in="" kidneys="" of="" ckom="" mice="" (figure="" 4d="" and="" supplemental="" figure="" 4).="" gene="" set="" enrichment="" analysis(gsea)="" performed="" on="" the="" kegg="" pathway="" database="" (release="" on="" december="" 11,="" 2020)showed="" downregulation="" of="" pathways="" related="" to="" the="" metabolism="" of="" pyruvate,="" glyoxylate="" and="" dicarboxylate,="" amino="" acids="" (arginine,="" proline,="" cysteine,="" methionine,="" tryptophan,="" alanine,="" and="" histidine),="" or="" glutathione(figure="" 4e="" and="" supplemental="" figure="" 5)="" and="" upregulation="" of="" pathways="" related="" to="" fatty="" acid="" synthesis="" and="" degradation,="" ppar="" signaling,="" and="" abc="" transporters="" (figure="" 4f="" and="" supplemental="" figure="" 6).="" no="" enrichment="" was="" found="" in="" pathways="" linked="" to="" inflammation="" or="" fibrosis.="" targeted="" analysis="" of="" several="" groups="" of="" functionally="" related="" genes="" that="" are="" not="" represented="" in="" kegg="" pathways="" revealed="" substantial="" changes="" in="" the="" expression="" of="" genes="" encoding="" enzymes="" involved="" in="" the="" peroxisomal="" metabolism="" of="" fatty="" acids="" (acsl1/3/4/6,="" acsvl1,="" acot3/4,="" acnat1,="" acox3,="" abcd3,="" ehhadh,="" and="" acaa1b),="" plasmalogen="" biosynthesis="" (far1="" and="" agps),="" bile="" acid="" synthesis(amacr="" and="" hsd17b4),="" and="" reactive="" oxygen="" species="" (ros)="" detoxification(cat="" and="" sod1)="" (supplemental="" figure="" 3).="" alterations="" were="" also="" noted="" in="" the="" expression="" of="" a="" large="" number="" of="" genes="" critical="" for="" membrane="" transport="" processes="" in="" the="" proximal="" tubule,="" thick="" ascending="" limb="" (tal),="" and="" distal="" nephron(supplemental="" figure="" 7="" and="" supplemental="" table="" 4).="" for="" instance,="" in="" the="" proximal="" tubule,="" we="" found="" a="" reduction="" in="" the="" expression="" of="" an="" aquaporin-1="" water="" channel(agp1);="" megalin="" (lrp2);="" phosphate="" transporter="" napi-2a="" (slc34al);="" urate="" transporters="" uratl="" (slc22a12),="" npt1/4="" (sic17al/3),="" and="" oat1="" (slc22a6);="" and="" numerous="" other="" organic="" anion="" and="" amino="" acid="" transporters.="" in="" the="" tal="" and="" distal="" nephron,="" there="" was="" a="" substantial="" increase="" in="" the="" expression="" of="" genes="" encoding="" proteins="" involved="" in="" sodium="" reabsorption:="" nkcc2="" (slc12a1),="" clc-kb="" (clcnkb),="" βenac="" (scnn1b)=""><0.05), and="" ncc="" (slc12a3)(fdr="">
<0.05. (d)="" enrichment="" analysis="" of="" a="" homemade="" gene="" set="" (based="" on="" the="" kegg="" pathway="" mmu04146)="" targeting="" 100="" transcripts="" related="" to="" peroxisomal="" functions,="" in="" ckom="" versus="" ctrlm="" mice.="" (e="" and="" f)="" scatter="" plot="" of="" the="" top="" 25="" most="" downregulated="" (e)="" or="" upregulated="" (f)="" metabolic="" pathways="" in="" ckom="" versus="" ctrlm="" mice,="" based="" on="" an="" untargeted="" gsea="" using="" a="" database="" of="" 543="" kegg="" metabolic="" pathways.="" pathways="" are="" sorted="" by="" their="" absolute="" normalized="" enrichment="" score.="" a="" significant="" pathway="" regulation="" can="" be="" considered="" when="" adjusted="" p="" value="" referred="" to="" as="" "q="" value"="" is=""><0.2" alt="Figure 4. Transcriptional reprogramming in the kidneys of cKO mice. (A and B) Volcano plot representing the relative transcriptional expression of all renal transcripts in cKOm versus Ctrlm (A) or cKOf versus Ctrlf (B). Transcripts depicted in blue are significantly downregulated while transcripts depicted in red are significantly upregulated. (C) Venn diagrams showing the number of transcripts significantly downregulated or upregulated in cKOm (in blue) or cKOf (in pink) mice versus Ctrl mice of the same sex. A significant transcript regulation is considered when the adjusted P value referred to as " fdr"="" is=""><0.05. (d)="" enrichment="" analysis="" of="" a="" homemade="" gene="" set="" (based="" on="" the="" kegg="" pathway="" mmu04146)="" targeting="" 100="" transcripts="" related="" to="" peroxisomal="" functions,="" in="" ckom="" versus="" ctrlm="" mice.="" (e="" and="" f)="" scatter="" plot="" of="" the="" top="" 25="" most="" downregulated="" (e)="" or="" upregulated="" (f)="" metabolic="" pathways="" in="" ckom="" versus="" ctrlm="" mice,="" based="" on="" an="" untargeted="" gsea="" using="" a="" database="" of="" 543="" kegg="" metabolic="" pathways.="" pathways="" are="" sorted="" by="" their="" absolute="" normalized="" enrichment="" score.="" a="" significant="" pathway="" regulation="" can="" be="" considered="" when="" adjusted="" p="" value="" referred="" to="" as="" "q="" value"="" is=""><0.2" width="480" height="320" border="0" vspace="0" style="width: 480px; height: 320px;">
Soti nan total de 852 metabolit detekte, 207 te montre diferans nan abondans nan ren sourit Ctrlm ak cKOm, ak 118 te montre abondans diferans nan ren nan sourit Ctrlf ak cKOf (Tablo Siplemantè 5, FDR).<5%). seventy-nine="" metabolites="" demonstrating="" differential="" abundance="" were="" common="" in="" both="" comparisons(figure="" 5a="" and="" supplemental="" table="" 6).among="" metabolites="" showing="" the="" most="" significant="" differences="" were="" plasmalogens="" and="" sphingomyelins(decreased="" abundance)and="" glutathione-related="" metabolites="" and="" dicarboxylic="" acids="" (increased="" abundance)="" in="" kidneys="" of="" both="" ckom="" and="" ckof="" mice="" (figure="" 5b="" and="" supplemental="" figure="" 8,="" respectively).="" global="" analysis="" of="" metabolome="" confirmed="" that="" plasmalogens="" and="" sphingomyelins="" constituted="" a="" majority="" of="" metabolites="" showing="" a="" decreased="" abundance="" in="" kidneys="" of="" both="" ckom="" and="" ckof="" mice="" (figure="" 5c).lcfa="" dicarboxylates,="" very="" long-chain="" fatty="" acyl-carnitines,="" phosphatidylcholines,="" and="" phosphatidylethanolamines="" were="" present="" among="" metabolites="" with="" increased="" abundance,="" in="" addition="" to="" glutathione-related="" metabolites="" and="" dicarboxylic="" acids(figure="">
<0.05. metabolites="" are="" identified="" by="" their="" biochemical="" name="" and="" sorted="" by="" related="" metabolisms="" and="" subclasses="" of="" metabolites.="" for="" each="" metabolite,="" the="" individual="" expression="" of="" 6="" ctrl="" and="" 6="" cko="" mice="" normalized="" between="" 0="" and="" 1="" and="" the="" log2="" -transformed="" mean="" fold="" change="" of="" expression="" (log2fc)="" in="" cko="" versus="" ctrl="" mice="" are="" given,="" for="" both="" sexes.="" for="" calculation="" of="" the="" mean="" fc="" of="" expression,="" missing="" values="" (depicted="" in="" gray)="" have="" been="" replaced="" by="" the="" minimum="" value="" of="" both="" genotypes="" from="" the="" same="" sex."="" alt="Figure 5. Remodeling of the renal metabolome in cKO mice. (A) Venn diagrams representing the number of detected renal metabolites showing a significantly decreased or increased abundance in cKOm (in blue) or cKOf (in pink) mice versus Ctrl mice of the same sex. (B) Volcano plot representing the relative abundance of all detected metabolites in kidneys of cKOm versus Ctrlm. Metabolites depicted with blue dots are significantly less abundant, and transcripts depicted with red dots are significantly more abundant in kidneys of cKOm mice as compared with Ctrlm mice. The names of some representative metabolites are depicted using colors shared for related metabolites. (C and D) Heatmaps of metabolites showing a significantly decreased (C) or increased (D) abundance in cKO mice of both sexes as compared with Ctrl mice. A significant difference of abundance is considered when adjusted P value from a 2-way ANOVA referred to as " fdr"="" is=""><0.05. metabolites="" are="" identified="" by="" their="" biochemical="" name="" and="" sorted="" by="" related="" metabolisms="" and="" subclasses="" of="" metabolites.="" for="" each="" metabolite,="" the="" individual="" expression="" of="" 6="" ctrl="" and="" 6="" cko="" mice="" normalized="" between="" 0="" and="" 1="" and="" the="" log2="" -transformed="" mean="" fold="" change="" of="" expression="" (log2fc)="" in="" cko="" versus="" ctrl="" mice="" are="" given,="" for="" both="" sexes.="" for="" calculation="" of="" the="" mean="" fc="" of="" expression,="" missing="" values="" (depicted="" in="" gray)="" have="" been="" replaced="" by="" the="" minimum="" value="" of="" both="" genotypes="" from="" the="" same="" sex."="" width="480" height="320" border="0" vspace="0" style="width: 480px; height: 320px;">
Analiz chemen konjwen (MetaboAnalyst 5.0) nan transkripsyon ak metabolit ki montre abondans ogmante oswa diminye nan ren sourit cKOm konpare ak sourit Ctrl idantifye 22 wout regilasyon downregulated ak 7upregulated (Figi 6, A ak B, respektivman; FDR).<0.1; supplemental="" table="" 7).="" nitrogen="" metabolism,="" pyruvate="" metabolism,="" glycolysis,="" and="" gluconeogenesis="" were="" identified="" among="" the="" downregulated="" pathways="" while="" fatty="" acid="" degradation="" was="" identified="" among="" the="" upregulated="" pathways="" (figure="" 6,="" a="" and="" b,="" respectively).="" glutathione="" metabolism="" and="" retinol="" metabolism="" were="" present="" among="" both="" up-and="" downregulated="" pathways="" (figure="" 6,="" a="" and="" b).="" detailed="" analysis="" of="" transcripts="" and="" metabolites="" related="" to="" glutathione="" metabolism="" identified="" 16="" transcripts="" and="" 3="" metabolites="" with="" decreased="" abundance="" in="" kidneys="" of="" ckom="" mice="" (figure="" 6,="" c="" and="" d,="" respectively),="" along="" with="" 6="" transcripts="" and="" 8="" metabolites="" exhibiting="" increased="" abundance(figure="" 6,="" e="" and="" f,="" respectively).="" the="" oxidized="" form="" of="" glutathione(gssg)was="" present="" in="" ckom="" but="" not="" in="" ctrlm="" mice,="" thereby="" suggesting="" oxidative="" stress="" in="" ckom="" mice(figure="">
<0.1. the="" size="" of="" each="" dot="" depends="" on="" the="" percentage="" of="" all="" transcripts="" and="" metabolites="" ("compounds")="" of="" the="" pathway="" that="" are="" significantly="" affected="" in="" ckom="" mice.="" (c="" and="" d)="" relative="" expression="" of="" glutathione-related="" transcripts="" (c)="" and="" metabolites="" (d)="" significantly="" less="" abundant="" (fdr="" <="" 0.05)="" in="" ckom="" mice="" as="" compared="" with="" ctrlm="" mice.="" (e="" and="" f)="" relative="" expression="" of="" glutathione-related="" transcripts="" (e)="" and="" metabolites="" (f)="" significantly="" more="" abundant="" (fdr="" <="" 0.05)="" in="" ckom="" mice="" as="" compared="" with="" ctrlm="" mice.="" individual="" values="" from="" 6="" ckom="" and="" 6="" ctrlm="" mice="" are="" depicted="" after="" transformation="" from="" raw="" individual="" data:="" values="" of="" metabolite="" abundance="" have="" been="" divided="" by="" the="" median="" value="" of="" both="" genotypes,="" while="" values="" of="" transcript="" expression="" from="" both="" genotypes="" have="" been="" normalized="" between="" 0="" and="" 1.="" box="" and="" whiskers="" represent="" the="" mean="" and="" the="" sem,="" respectively.="" nd,="" not="" detected."="" alt="Figure 6. Joint analysis of transcriptional and metabolic changes in cKOm mice. (A and B) Scatter plot of significantly downregulated (A) or upregulated (B) metabolic pathways, based on a joint pathway analysis of both regulated transcripts and modulated metabolites in kidneys of cKOm versus Ctrlm mice. Pathways are sorted by their absolute impact, and a significant pathway regulation is considered when adjusted P value referred to as " fdr"="" is=""><0.1. the="" size="" of="" each="" dot="" depends="" on="" the="" percentage="" of="" all="" transcripts="" and="" metabolites="" ("compounds")="" of="" the="" pathway="" that="" are="" significantly="" affected="" in="" ckom="" mice.="" (c="" and="" d)="" relative="" expression="" of="" glutathione-related="" transcripts="" (c)="" and="" metabolites="" (d)="" significantly="" less="" abundant="" (fdr="" <="" 0.05)="" in="" ckom="" mice="" as="" compared="" with="" ctrlm="" mice.="" (e="" and="" f)="" relative="" expression="" of="" glutathione-related="" transcripts="" (e)="" and="" metabolites="" (f)="" significantly="" more="" abundant="" (fdr="" <="" 0.05)="" in="" ckom="" mice="" as="" compared="" with="" ctrlm="" mice.="" individual="" values="" from="" 6="" ckom="" and="" 6="" ctrlm="" mice="" are="" depicted="" after="" transformation="" from="" raw="" individual="" data:="" values="" of="" metabolite="" abundance="" have="" been="" divided="" by="" the="" median="" value="" of="" both="" genotypes,="" while="" values="" of="" transcript="" expression="" from="" both="" genotypes="" have="" been="" normalized="" between="" 0="" and="" 1.="" box="" and="" whiskers="" represent="" the="" mean="" and="" the="" sem,="" respectively.="" nd,="" not="" detected."="" width="480" height="320" border="0" vspace="0" style="width: 480px; height: 320px;">
Defi manje ki gen anpil grès pa mennen nan estrès oksidatif adisyonèl nan ckKOmice. Chanjman nan chemen ki gen rapò ak glutatyon, rediksyon nan plasmalogens, ak diminye ekspresyon de katalaz sijere estrès oksidatif ak / oswa reyajman nan diferan sistèm defans antioksidan nan ren nan sourit cKOm. Pou teste ipotèz sa yo, nou te defye sourit cKOm ak yon rejim ki gen anpil grès (HFD) pou 4 semèn. Defi sa a pa te lakòz albuminuria men ogmante volim urin ak eskresyon urin nan kalsyòm ak urat. Pa gen okenn diferans obsève nan paramèt plasma teste, ki gen ladan kalkemi ak uricemi (Tablo Siplemantè 8). Pa gen okenn chanjman mòfolojik brit oswa akimilasyon lipid yo te detekte nan ren yo nan sourit HFD-challenged Ctrlm ak cKOm (Figi 7A). Kapasite total ak nonenzymatic antioksidan jan yo evalye Trolox esè (Figi 7B) osi byen ke nivo tisi nan malondialdehyde, yon makè nan peroksidasyon asid gra poliensature (Figi 7C), yo pa t diferan ant tretman ak jenotip. Abondans pwodwi peroksidasyon lipid 4-hydroxynonenal (4-HNE) te ogmante nan sourit Ctrlm ki te trete ak HFD an konparezon ak sourit Ctrl nan rejim kontwòl la, men pa te obsève diferans nan sourit cKOm trete ak 2 a. alimantasyon (Figi 7D).
