{"id":6916,"date":"2021-04-18T09:12:39","date_gmt":"2021-04-18T07:12:39","guid":{"rendered":"https:\/\/www.fipwarriors.eu\/?p=6916"},"modified":"2023-03-29T09:50:04","modified_gmt":"2023-03-29T07:50:04","slug":"mozna-antivirusova-aktivita-kyseliny-5-aminolevulovej-pri-infekcii-virusom-infekcnej-peritonitidy-maciek","status":"publish","type":"post","link":"https:\/\/www.fipwarriors.eu\/en\/mozna-antivirusova-aktivita-kyseliny-5-aminolevulovej-pri-infekcii-virusom-infekcnej-peritonitidy-maciek\/","title":{"rendered":"Possible Antiviral Activity of 5-Aminolevulinic Acid in Feline Infectious Peritonitis Virus (Feline Coronavirus) Infection"},"content":{"rendered":"<p class=\"has-small-font-size wp-block-paragraph\">Tomomi Takano, Kumi Satoh, and Tomoyoshi Doki<br>Origin&aacute;lny &#269;l&aacute;nok: <a href=\"https:\/\/www.ncbi.nlm.nih.gov\/pmc\/articles\/PMC7903937\/\" target=\"_blank\" rel=\"noreferrer noopener nofollow\">Possible Antiviral Activity of 5-Aminolevulinic Acid in Feline Infectious Peritonitis Virus (Feline Coronavirus) Infection<\/a>; 10.2.2021;Preklad 18.4.2021<\/p>\n\n\n\n<h2 class=\"lm-underscore wp-block-heading\">Abstrakt<\/h2>\n\n\n\n<p class=\"wp-block-paragraph\">Infek&#269;n&aacute; peritonit&iacute;da ma&#269;iek (<a class=\"glossaryLink\" aria-describedby=\"tt\" data-cmtooltip=\"&lt;div class=glossaryItemBody&gt;Infek&#269;n&aacute; peritonit&iacute;da ma&#269;iek (angl. Feline infectious peritonitis, zau&#382;&iacute;van&aacute; skratka FIP) je infek&#269;n&eacute; ochorenie ma&#269;iek sp&ocirc;sobuj&uacute;ce smr&#357;. N&aacute;zov je mierne zav&aacute;dzaj&uacute;ci, preto&#382;e nejde o z&aacute;pal pobru&scaron;nice (&#269;i&#382;e peritonea), ale o z&aacute;pal ciev (vaskulit&iacute;da). Klinick&eacute; pr&iacute;znaky FIP sa v&scaron;ak &#269;asto prejavuj&uacute; vo forme z&aacute;palu pobru&scaron;nice. V&iacute;rus FIP nap&aacute;da v&scaron;ak aj pohrudnicu &ndash; z toho d&ocirc;vodu je niekedy choroba ozna&#269;ovan&aacute; tie&#382; ako Infek&#269;n&aacute; ma&#269;acia polyserozit&iacute;da (Feline infectious polyserositis).&lt;\/div&gt;\" href=\"https:\/\/www.fipwarriors.eu\/en\/glossary\/fip\/\" data-mobile-support=\"0\" data-gt-translate-attributes='[{\"attribute\":\"data-cmtooltip\", \"format\":\"html\"}]' tabindex=\"0\" role=\"link\">FIP<\/a>) je &#382;ivot ohrozuj&uacute;ce infek&#269;n&eacute; ochorenie ma&#269;iek sp&ocirc;soben&eacute; virulentn&yacute;m ma&#269;ac&iacute;m koronav&iacute;rusom (v&iacute;rus FIP: FIPV). Na lie&#269;bu FIP sa ned&aacute;vno objavilo nieko&#318;ko &uacute;&#269;inn&yacute;ch antivirot&iacute;k, ale mnoh&eacute; z nich nie s&uacute; dostupn&eacute; pre praktick&eacute; pou&#382;itie. Kyselina 5-aminolevulov&aacute; (5-ALA) je n&iacute;zkomolekul&aacute;rna aminokyselina syntetizovan&aacute; v rastlinn&yacute;ch a &#382;ivo&#269;&iacute;&scaron;nych bunk&aacute;ch. 5-ALA m&ocirc;&#382;e by&#357; syntetizovan&aacute; vo ve&#318;kom mno&#382;stve a je &scaron;iroko pou&#382;&iacute;van&aacute; v lek&aacute;rskych a po&#318;nohospod&aacute;rskych odvetviach. Dospeli sme k predpokladu, &#382;e 5-ALA inhibuje infekciu FIPV. Preto sme vyhodnotili jej antiv&iacute;rusov&uacute; aktivitu proti FIPV v cel&yacute;ch plodov&yacute;ch 4-bunk&aacute;ch felis catus a v prim&aacute;rnych ma&#269;ac&iacute;ch makrof&aacute;goch. FIPV infekcia bola v&yacute;znamne inhibovan&aacute; 250 &mu;M 5-ALA. Na&scaron;a &scaron;t&uacute;dia nazna&#269;ila, &#382;e 5-ALA je pou&#382;ite&#318;n&aacute; pre lie&#269;bu a prevenciu infekcie FIPV.<\/p>\n\n\n\n<p class=\"wp-block-paragraph\"><strong>K&#318;&uacute;&#269;ov&eacute; slov&aacute;: <\/strong>FIP, koronav&iacute;rus, kyselina 5-aminolevulov&aacute;, antiv&iacute;rusov&eacute; lie&#269;ivo, ma&#269;ka<\/p>\n\n\n\n<h2 class=\"lm-underscore wp-block-heading\">&Uacute;vod<\/h2>\n\n\n\n<p class=\"wp-block-paragraph\">Infek&#269;n&aacute; peritonit&iacute;da ma&#269;iek (FIP) je &#382;ivot ohrozuj&uacute;ce infek&#269;n&eacute; ochorenie sp&ocirc;soben&eacute; ma&#269;ac&iacute;m koronav&iacute;rusom (FCoV) u dom&aacute;cich a vo&#318;ne &#382;ij&uacute;cich druhov Felidae. FCoV je u ma&#269;iek vysoko roz&scaron;&iacute;ren&yacute; po celom svete. FCoV je obalen&yacute; jednovl&aacute;knov&yacute; pozit&iacute;vny <a class=\"glossaryLink\" aria-describedby=\"tt\" data-cmtooltip=\"&lt;div class=glossaryItemBody&gt;Ribonukleov&aacute; kyselina. Polymern&iacute; makromolekula, jej&iacute;&#382; z&aacute;kladn&iacute;mi stavebn&iacute;mi jednotkami jsou ribonukleotidy. Je sou&#269;&aacute;st&iacute; v&scaron;ech bun&#283;k a slou&#382;&iacute; k p&#345;enosu informac&iacute; z DNA d&aacute;le do bu&#328;ky. Existuje mnoho druh&#367; RNA s r&#367;zn&yacute;mi funkcemi. Mezi nejzn&aacute;mn&#283;j&scaron;&iacute; pat&#345;&iacute; mRNA (medi&aacute;torov&aacute;), tRNA (transferov&aacute;) a rRNA (ribozom&aacute;ln&iacute;).&lt;\/div&gt;\" href=\"https:\/\/www.fipwarriors.eu\/en\/glossary\/rna\/\" data-mobile-support=\"0\" data-gt-translate-attributes='[{\"attribute\":\"data-cmtooltip\", \"format\":\"html\"}]' tabindex=\"0\" role=\"link\">RNA<\/a> v&iacute;rus. Tento v&iacute;rus patr&iacute; do rodu Alphacoronavirus z pod&#269;e&#318;ade Orthocoronavirinae z &#269;e&#318;ade Coronaviridae (1). FCoV je sa del&iacute; na dva s&eacute;rotypy pod&#318;a aminokyselinovej sekvencie spike prote&iacute;nu (S), s&eacute;rotyp I FCoV a s&eacute;rotyp II FCoV (2). S&eacute;rologick&eacute; a genetick&eacute; v&yacute;skumy odhalili, &#382;e FCoV typu I je dominantn&yacute; na celom svete (3&ndash;5). FCoV sa &scaron;&iacute;ri hlavne fek&aacute;lno-or&aacute;lnym prenosom (6). V&auml;&#269;&scaron;ina ma&#269;iek infikovan&yacute;ch FCoV je subklinick&yacute;ch. V S prote&iacute;ne sa v&scaron;ak vyskytlo nieko&#318;ko mut&aacute;ci&iacute;, ktor&eacute; viedli k v&yacute;voju virulentn&eacute;ho typu naz&yacute;van&eacute;ho v&iacute;rus ma&#269;acej infek&#269;nej peritonit&iacute;dy (FIPV) (7, 8). Charakteristick&yacute;m patologick&yacute;m n&aacute;lezom FIP u ma&#269;iek je s&eacute;rov&aacute; tekutina v peritone&aacute;lnych a pleur&aacute;lnych dutin&aacute;ch a pyogranulomat&oacute;zne l&eacute;zie v nieko&#318;k&yacute;ch org&aacute;noch (9).<\/p>\n\n\n\n<p class=\"wp-block-paragraph\">FIP je imunitne sprostredkovan&aacute; a &#357;a&#382;ko lie&#269;ite&#318;n&aacute; v&iacute;rusov&aacute; infekcia. Ned&aacute;vno bolo publikovan&yacute;ch nieko&#318;ko &uacute;&#269;inn&yacute;ch antivirot&iacute;k na lie&#269;bu FIP (10, 11), mnoh&eacute; z nich v&scaron;ak nie s&uacute; dostupn&eacute; pre praktick&eacute; pou&#382;itie. Niektor&eacute; lieky proti FCoV, ako napr&iacute;klad itrakonazol, s&uacute; dostupn&eacute; v kinik&aacute;ch pre zvierat&aacute;, ale ich lie&#269;ebn&eacute; &uacute;&#269;inky s&uacute; obmedzen&eacute; (12). Preto&#382;e FIP je chronick&eacute; a syst&eacute;mov&eacute; ochorenie, je &#357;a&#382;k&eacute; dosiahnu&#357; klinick&uacute; remisiu. Preto je &#382;iaduce, aby terapeutick&eacute; lieky na FIP mali nasleduj&uacute;ce vlastnosti: (1) M&aacute;lo ved&#318;aj&scaron;&iacute;ch &uacute;&#269;inkov pre ma&#269;ky, (2) n&iacute;zka cena a (3) n&iacute;zka mutagen&eacute;za patog&eacute;nov.<\/p>\n\n\n\n<p class=\"wp-block-paragraph\">Kyselina 5-aminolevulov&aacute; (5-ALA) je n&iacute;zkomolekul&aacute;rna aminokyselina syntetizovan&aacute; v rastlinn&yacute;ch a &#382;ivo&#269;&iacute;&scaron;nych bunk&aacute;ch (13, 14). Je medziproduktom v biosynt&eacute;ze tetrapyrolu. Preto&#382;e 5-ALA je vysoko rozpustn&aacute; vo vode a m&aacute; n&iacute;zku cytotoxicitu, je &scaron;iroko pou&#382;&iacute;van&aacute; v lek&aacute;rskych a po&#318;nohospod&aacute;rskych odvetviach(15). Bolo publikovan&yacute;ch nieko&#318;ko &scaron;t&uacute;di&iacute; o &uacute;&#269;inkoch 5-ALA na infek&#269;n&eacute; choroby. Suzuki a kol. uviedli, &#382;e ke&#271; sa 5-ALA a i&oacute;n &#382;eleza pod&aacute;vali or&aacute;lne my&scaron;iam infikovan&yacute;m parazitom sp&ocirc;sobuj&uacute;com mal&aacute;riu (<em>Plasmodium yoelii<\/em>), my&scaron;i pre&#382;ili (16). Na druhej strane s&uacute; jeho &uacute;&#269;inky na v&iacute;rusov&uacute; infekciu nejasn&eacute;.<\/p>\n\n\n\n<p class=\"wp-block-paragraph\">Vo veterin&aacute;rnom lek&aacute;rstve bola fotodynamick&aacute; terapia (PDT) s pou&#382;it&iacute;m 5-ALA sk&uacute;man&aacute; pri lie&#269;be n&aacute;dorov u psov (17), ale pod&#318;a na&scaron;ich vedomost&iacute; nebol sk&uacute;man&yacute; vplyv 5-ALA na infek&#269;n&eacute; choroby zvierat. Sk&uacute;mali sme, &#269;i 5-ALA m&ocirc;&#382;e by&#357; aplikovan&aacute; ako anti-FCoV liek <em><a class=\"glossaryLink\" aria-describedby=\"tt\" data-cmtooltip=\"&lt;div class=glossaryItemBody&gt;in vitro - lat. &amp;quot;V skle&amp;quot; tj. V sk&uacute;mavke, za umel&yacute;ch, laborat&oacute;rnych podmienok. Napr. pri testovan&iacute; &uacute;&#269;inkov chemick&yacute;ch l&aacute;tok na &#382;iv&yacute; organizmus s&uacute; &uacute;&#269;inky sk&uacute;man&eacute; najprv na kult&uacute;re buniek pestovan&yacute;ch v sk&uacute;mavke. In&yacute;m pr&iacute;kladom je oplodnenie in vitro &amp;quot;die&#357;a zo sk&uacute;mavky&amp;quot;, IVF. Porov. in vivo lat. in v; VITRUM sklo&lt;\/div&gt;\" href=\"https:\/\/www.fipwarriors.eu\/en\/glossary\/in-vitro\/\" data-mobile-support=\"0\" data-gt-translate-attributes='[{\"attribute\":\"data-cmtooltip\", \"format\":\"html\"}]' tabindex=\"0\" role=\"link\">in vitro<\/a><\/em>.<\/p>\n\n\n\n<h2 class=\"lm-underscore wp-block-heading\">Materi&aacute;ly a met&oacute;dy<\/h2>\n\n\n\n<h3 class=\"wp-block-heading\">Bunkov&eacute; kult&uacute;ry, zvierat&aacute; a v&iacute;rusy<\/h3>\n\n\n\n<p class=\"wp-block-paragraph\">Ma&#269;acie plodov&eacute; 4-bunky Felis catus (fcwf)-4 (l&aacute;skavo dodan&eacute; Dr. MC Horzinkem z Universiteit Utrecht) boli vypestovan&eacute; v Eagles &sbquo;MEM obsahuj&uacute;com 50% m&eacute;dia L-15 Leibovitz, 5% fet&aacute;lne te&#318;acie <a class=\"glossaryLink\" aria-describedby=\"tt\" data-cmtooltip=\"&lt;div class=glossaryItemBody&gt;&amp;lt;!-- wp:paragraph --&amp;gt;Krvn&eacute; s&eacute;rum je &#382;ltkast&aacute;, tekut&aacute; a nebunkov&aacute; zlo&#382;ka krvi, ktor&aacute; vznik&aacute; po vyzr&aacute;&#382;an&iacute; krvi a n&aacute;slednom odstr&aacute;nen&iacute; krvn&eacute;ho kol&aacute;&#269;a mechanicky a pomocou centrif&uacute;gy. Zlo&#382;en&iacute;m zodpoved&aacute; krvnej plazme, av&scaron;ak na rozdiel od plazmy neobsahuje fibrinog&eacute;n a &#271;al&scaron;ie zr&aacute;&#382;acie faktory krvi.&amp;lt;br\/&amp;gt;&amp;lt;!-- \/wp:paragraph --&amp;gt;&amp;lt;!-- wp:paragraph --&amp;gt;Krv, ktor&aacute; sa odober&aacute; na pr&iacute;pravu s&eacute;ra, sa odober&aacute; do oby&#269;ajn&yacute;ch sk&uacute;maviek alebo strieka&#269;iek bez pridania ak&yacute;chko&#318;vek antikoagulantov (napr. hepar&iacute;n, EDTA). V klinickej a laborat&oacute;rnej praxi sa &#269;asto pou&#382;&iacute;va ozna&#269;enie s&eacute;rum.&amp;lt;br\/&amp;gt;&amp;lt;!-- \/wp:paragraph --&amp;gt;&lt;\/div&gt;\" href=\"https:\/\/www.fipwarriors.eu\/en\/glossary\/krvne-serum\/\" data-mobile-support=\"0\" data-gt-translate-attributes='[{\"attribute\":\"data-cmtooltip\", \"format\":\"html\"}]' tabindex=\"0\" role=\"link\">s&eacute;rum<\/a> (FCS), 100 U\/ml penicil&iacute;nu a 100 &mu;g\/ml streptomyc&iacute;nu. Udr&#382;iavacie m&eacute;dium malo rovnak&eacute; zlo&#382;enie ako rastov&eacute; m&eacute;dium s v&yacute;nimkou koncentr&aacute;cie FCS (2%). Pre prim&aacute;rne <a class=\"glossaryLink\" aria-describedby=\"tt\" data-cmtooltip=\"&lt;div class=glossaryItemBody&gt;Makrof&aacute;g je bu&#328;ka p&#345;irozen&eacute; imunity, kter&aacute; hraje velmi d&#367;le&#382;itou roli v imunitn&iacute; reakci. Jedn&aacute; se o z&aacute;stupce mononukle&aacute;r&#367;, tj. bun&#283;k s jedn&iacute;m, nesegmentovan&yacute;m j&aacute;drem.Makrof&aacute;g vznik&aacute; p&#345;em&#283;nou z monocyt&#367;. Ty jsou tvo&#345;eny v kostn&iacute; d&#345;eni z kmenov&eacute; hemopoetick&eacute; bu&#328;ky a jsou vyplavov&aacute;ny do krevn&iacute;ho ob&#283;hu. Monocyty koluj&iacute; v krvi asi 8 hodin, pot&eacute; vstupuj&iacute; do tk&aacute;n&iacute; a tam se m&#283;n&iacute; na makrof&aacute;gy. Tk&aacute;&#328;ov&eacute; makrof&aacute;gy pak vykazuj&iacute; &#269;etnou heterogenitu v z&aacute;vislosti na tk&aacute;ni. Mezi tk&aacute;&#328;ov&eacute; makrof&aacute;gy pat&#345;&iacute; Kupfferovy bu&#328;ky, histiocyty, osteoklasty &#269;i mikroglie.Aby se monocyt p&#345;em&#283;nil na makrof&aacute;g, mus&iacute; prob&#283;hnout n&#283;kolik krok&#367;. Zv&#283;t&scaron;uje se velikost bu&#328;ky, zvy&scaron;uje se po&#269;et lysozom&#367;, receptor&#367; pro imunoglobulin IgG a zvy&scaron;uje se jeho schopnost fagocyt&oacute;zy.Z&aacute;kladn&iacute; funkc&iacute; makrof&aacute;gu je fagocyt&oacute;za. K dal&scaron;&iacute;m funkc&iacute;m pat&#345;&iacute; prezentace antigenu T-lymfocyt&#367;m, &#345;&iacute;zen&iacute; hemopo&eacute;zy, hemost&aacute;zy a hojen&iacute; ran, regulace z&aacute;n&#283;tu, destrukce mikroorganism&#367;, odstra&#328;ov&aacute;n&iacute; mrtv&yacute;ch bun&#283;k a cytotoxick&aacute; reakce.&lt;\/div&gt;\" href=\"https:\/\/www.fipwarriors.eu\/en\/glossary\/makrofag\/\" data-mobile-support=\"0\" data-gt-translate-attributes='[{\"attribute\":\"data-cmtooltip\", \"format\":\"html\"}]' tabindex=\"0\" role=\"link\">makrof&aacute;gy<\/a> boli vybran&eacute; ma&#269;acie prim&aacute;rne makrof&aacute;gy. Ma&#269;acie alveol&aacute;rne makrof&aacute;gy sa z&iacute;skali zo &scaron;tyroch ma&#269;iek bez &scaron;pecifick&yacute;ch patog&eacute;nov (SPF) vo veku 3 &ndash; 5 rokov bronchoalveol&aacute;rnou lav&aacute;&#382;ou s Hankov&yacute;m vyv&aacute;&#382;en&yacute;m so&#318;n&yacute;m roztokom. Ma&#269;acie prim&aacute;rne makrof&aacute;gy sa udr&#382;iavali v D-MEM doplnenom o 10% FCS, 100 U\/ml penicil&iacute;nu a 100 &mu;g\/ml streptomyc&iacute;nu. SPF ma&#269;ky sa udr&#382;iavali v izolovanom zariaden&iacute; s regulovanou teplotou. Pokus na zvierat&aacute;ch bol schv&aacute;len&yacute; prezidentom Kitasato University na z&aacute;klade rozhodnutia In&scaron;titucion&aacute;lneho v&yacute;boru pre starostlivos&#357; a pou&#382;itie zvierat Kitasato University (18-050) a uskuto&#269;nen&yacute; v s&uacute;lade s Guidelines for Animal Experiments of Kitasato University. Ve&#318;kosti vzoriek boli ur&#269;en&eacute; na z&aacute;klade na&scaron;ej predch&aacute;dzaj&uacute;cej &scaron;t&uacute;die a bol pou&#382;it&yacute; minim&aacute;lny po&#269;et ma&#269;iek. Kme&#328; FCoV KU-2 typu I (FIPV-I KU-2) bol izolovan&yacute; v na&scaron;om laborat&oacute;riu. FCoV-II 79-1146 l&aacute;skavo poskytol Dr. M. C. Horzinek z Utrechtskej univerzity. Tieto v&iacute;rusy boli kultivovan&eacute; v bunk&aacute;ch fcwf-4 pri 37&deg;C.<\/p>\n\n\n\n<h3 class=\"wp-block-heading\">Zl&uacute;&#269;eniny<\/h3>\n\n\n\n<p class=\"wp-block-paragraph\">5-ALA a citr&aacute;t &#382;eleznat&yacute; sodn&yacute; (SFC) poskytla spolo&#269;nos&#357; Neopharma Japan (Tokio, Japonsko). 5-ALA a SFC sa rozpustili v kultiva&#269;nom m&eacute;diu pri 200 mM a 50 mM. Roztok SFC sa pou&#382;il ako rozp&uacute;&scaron;&#357;adlo 5-ALA. V de&#328; experimentov sa tieto zl&uacute;&#269;eniny rozriedili na po&#382;adovan&eacute; koncentr&aacute;cie v kultiva&#269;nom m&eacute;diu.<\/p>\n\n\n\n<h3 class=\"wp-block-heading\">Cytotoxick&eacute; &uacute;&#269;inky zl&uacute;&#269;en&iacute;n<\/h3>\n\n\n\n<p class=\"wp-block-paragraph\">Bunky fcwf-4 sa nasadili na 96-jamkov&eacute; do&scaron;ti&#269;ky. Zl&uacute;&#269;eniny boli nasaden&eacute; do jamiek trojmo. Po inkub&aacute;cii v trvan&iacute; 96 hod&iacute;n sa supernatanty kult&uacute;ry odstr&aacute;nili, pridal sa roztok WST-8 (Kishida Chemical, Osaka, Japonsko) a bunky sa vr&aacute;tili do inkub&aacute;tora na 1 hodinu. Absorbancia vyprodukovan&eacute;ho formazanu sa merala pri 450 nm pomocou 96-jamkovej spektrofotometrickej &#269;&iacute;ta&#269;ky do&scaron;ti&#269;iek, pod&#318;a doporu&#269;enia v&yacute;robcu. Percento &#382;ivotaschopnosti buniek bolo vypo&#269;&iacute;tan&eacute; pomocou nasleduj&uacute;ceho vzorca: &#381;ivotaschopnos&#357; buniek (%) = [(OD buniek neo&scaron;etren&yacute;ch zl&uacute;&#269;eninou &ndash; buniek o&scaron;etren&yacute;ch zl&uacute;&#269;eninou)\/(OD buniek neo&scaron;etren&yacute;ch zl&uacute;&#269;eninou)]x100. 50% koncentr&aacute;cia cytotoxicity (CC<sub>50<\/sub> ) bola definovan&aacute; ako cytotoxick&aacute; koncentr&aacute;cia ka&#382;dej zl&uacute;&#269;eniny, ktor&aacute; zn&iacute;&#382;ila absorbanciu o&scaron;etren&yacute;ch buniek na 50% v porovnan&iacute; s absorpciou neo&scaron;etren&yacute;ch buniek.<\/p>\n\n\n\n<h3 class=\"wp-block-heading\">Antiv&iacute;rusov&eacute; &uacute;&#269;inky 5-ALA<\/h3>\n\n\n\n<p class=\"wp-block-paragraph\">Konfluentn&eacute; monovrstvy buniek fcwf-4 sa kultivovali v m&eacute;diu so zl&uacute;&#269;eninami alebo bez nich v uveden&yacute;ch koncentr&aacute;ci&aacute;ch na 24-jamkov&yacute;ch viac&scaron;tancov&yacute;ch platniach pri 37&deg;C po&#269;as 24 alebo 48 hod&iacute;n. Bunky sa premyli a v&iacute;rus (MOI 0,01) sa adsorboval do buniek pri teplote 37&deg;C po&#269;as 1 hodiny. Po premyt&iacute; sa bunky kultivovali v 1,5% karboxymetylcelul&oacute;ze (CMC)-MEM alebo MEM so zl&uacute;&#269;eninami alebo bez nich. V pr&iacute;pade buniek kultivovan&yacute;ch v CMC-MEM boli bunkov&eacute; monovrstvy inkubovan&eacute; pri teplote 37&deg;C po dobu 48 hod&iacute;n, fixovan&eacute; a zafarben&eacute; 1% roztokom kry&scaron;talickej fialovej obsahuj&uacute;cim 10% pufrovan&eacute;ho formal&iacute;nu a n&aacute;sledne boli spo&#269;&iacute;tan&eacute; v&yacute;sledn&eacute; plaky. Percento zn&iacute;&#382;enia alebo zv&yacute;&scaron;enia plakov bolo vypo&#269;&iacute;tan&eacute; pomocou nasleduj&uacute;ceho vzorca: Percento redukcie plakov (%) = [(po&#269;et plakov buniek o&scaron;etren&yacute;ch zl&uacute;&#269;eninou)\/(po&#269;et plakov buniek neo&scaron;etren&yacute;ch zl&uacute;&#269;eninou)]x100. EC<sub>50<\/sub> bola definovan&aacute; ako &uacute;&#269;inn&aacute; koncentr&aacute;cia zl&uacute;&#269;en&iacute;n, ktor&aacute; zn&iacute;&#382;ila <a class=\"glossaryLink\" aria-describedby=\"tt\" data-cmtooltip=\"&lt;div class=glossaryItemBody&gt;&amp;lt;strong&amp;gt;Titr&amp;lt;\/strong&amp;gt; &ndash; v s&eacute;rologii pojem u&#382;&iacute;van&yacute; k vyj&aacute;d&#345;en&iacute; stupn&#283; z&#345;ed&#283;n&iacute; vy&scaron;et&#345;ovan&eacute;ho krevn&iacute;ho s&eacute;ra, p&#345;i n&#283;m&#382; jsou s&eacute;rov&eacute; protil&aacute;tky je&scaron;t&#283; schopny viditeln&#283; reagovat s p&#345;&iacute;slu&scaron;n&yacute;m antigenem bakteri&aacute;ln&iacute;m, virov&yacute;m, pl&iacute;s&#328;ov&yacute;m apod.. &#268;&iacute;m vy&scaron;&scaron;&iacute; je t., t&iacute;m v&iacute;ce lze z&#345;edit s&eacute;rum a je&scaron;t&#283; doc&iacute;lit p&#345;&iacute;slu&scaron;n&eacute; reakce. Z toho lze usuzovat na obsah a mno&#382;stv&iacute; protil&aacute;tek v krvi a t&iacute;m i na infekci a jej&iacute; intenzitu.&lt;\/div&gt;\" href=\"https:\/\/www.fipwarriors.eu\/en\/glossary\/titer\/\" data-mobile-support=\"0\" data-gt-translate-attributes='[{\"attribute\":\"data-cmtooltip\", \"format\":\"html\"}]' tabindex=\"0\" role=\"link\">titer<\/a> v&iacute;rusu v supernatante kult&uacute;ry infikovan&yacute;ch buniek na 50% v porovnan&iacute; s titrom v&iacute;rusovej kontroly. V pr&iacute;pade buniek kultivovan&yacute;ch v MEM sa kultiva&#269;n&eacute; supernatanty zozbierali 48 hod&iacute;n po infekcii a titre v&iacute;rusu sa merali pomocou testu TCID<sub>50<\/sub>.<\/p>\n\n\n\n<p class=\"wp-block-paragraph\">Prim&aacute;rne ma&#269;acie makrof&aacute;gy sa kultivovali v m&eacute;diu so zl&uacute;&#269;eninami alebo bez nich v uveden&yacute;ch koncentr&aacute;ci&aacute;ch na 24-jamkov&yacute;ch viacn&aacute;sobn&yacute;ch do&scaron;ti&#269;k&aacute;ch pri 37&deg;C po dobu 48 hod&iacute;n. Po premyt&iacute; PBS sa nechal FIPV 79-1146 (1 x 10<sup>4<\/sup> TCID<sub>50<\/sub>) adsorbova&#357; do buniek pri 37&deg;C s 5% CO<sub>2<\/sub> po dobu 1 hodiny. Po premyt&iacute; PBS sa bunky kultivovali v m&eacute;diu a supernatanty sa zhroma&#382;dili. Titre v&iacute;rusu sa merali pomocou testu TCID<sub>50<\/sub>.<\/p>\n\n\n\n<h3 class=\"wp-block-heading\">&Scaron;tatistick&aacute; anal&yacute;za<\/h3>\n\n\n\n<p class=\"wp-block-paragraph\">&Uacute;daje iba z dvoch skup&iacute;n sa analyzovali pomocou Studentovho t-testu (Welchov t-test) a &uacute;daje viacer&yacute;ch skup&iacute;n sa analyzovali jednosmernou ANOVA nasledovanou Tukeyho testom. Za v&yacute;znamn&uacute; sa pova&#382;ovala hodnota P &lt;0,05.<\/p>\n\n\n\n<h2 class=\"lm-underscore wp-block-heading\">V&yacute;sledky<\/h2>\n\n\n\n<h2 class=\"wp-block-heading\">Cytotoxick&eacute; a antiv&iacute;rusov&eacute; &uacute;&#269;inky 5-ALA<\/h2>\n\n\n\n<p class=\"wp-block-paragraph\">Pre stanovenie netoxickej koncentr&aacute;cie 5-ALA proti bunk&aacute;m fcwf-4 sa vykonal test cytotixicity (obr&aacute;zok 1). Viac ako 75% buniek fcwf-4 pre&#382;ilo v pr&iacute;tomnosti 1 000 &mu;M 5-ALA (maxim&aacute;lna koncentr&aacute;cia v tomto experimente). Vehikulum nevykazuje &#382;iadne cytotoxick&eacute; &uacute;&#269;inky na bunky fcwf-4.<\/p>\n\n\n\n<div class=\"wp-block-media-text alignwide is-stacked-on-mobile is-vertically-aligned-top lm-back\"><figure class=\"wp-block-media-text__media\"><img loading=\"lazy\" decoding=\"async\" width=\"709\" height=\"719\" src=\"https:\/\/www.fipwarriors.eu\/wp-content\/uploads\/2021\/04\/5ala1.jpg\" alt=\"\" class=\"wp-image-6933 size-full\" srcset=\"https:\/\/www.fipwarriors.eu\/wp-content\/uploads\/2021\/04\/5ala1.jpg 709w, https:\/\/www.fipwarriors.eu\/wp-content\/uploads\/2021\/04\/5ala1-296x300.jpg 296w, https:\/\/www.fipwarriors.eu\/wp-content\/uploads\/2021\/04\/5ala1-592x600.jpg 592w, https:\/\/www.fipwarriors.eu\/wp-content\/uploads\/2021\/04\/5ala1-12x12.jpg 12w, https:\/\/www.fipwarriors.eu\/wp-content\/uploads\/2021\/04\/5ala1-100x100.jpg 100w, https:\/\/www.fipwarriors.eu\/wp-content\/uploads\/2021\/04\/5ala1-350x355.jpg 350w, https:\/\/www.fipwarriors.eu\/wp-content\/uploads\/2021\/04\/5ala1-64x65.jpg 64w\" sizes=\"auto, (max-width: 709px) 100vw, 709px\"\/><\/figure><div class=\"wp-block-media-text__content\">\n<p class=\"has-small-font-size wp-block-paragraph\"><strong><em>Obr&aacute;zok 1.<\/em><\/strong><\/p>\n\n\n\n<p class=\"has-small-font-size wp-block-paragraph\"><em>Cytotoxick&eacute; &uacute;&#269;inky 5-ALA na bunky fcwf-4. &#381;ivotaschopnos&#357; buniek fcwf-4 sa merala pomocou testu WST-8. &#268;ierny kr&uacute;&#382;ok: 5-ALA. Biely kr&uacute;&#382;ok: Vehikulum (SFC). Vehikulum (kontroln&eacute; rozp&uacute;&scaron;&#357;adlo) bolo pri ka&#382;dom postupnom rieden&iacute; rovnak&eacute; ako vehikulum v roztoku 5-ALA. V&yacute;sledky s&uacute; uveden&eacute; ako priemer &plusmn; SE. &Uacute;daje predstavuj&uacute; tri nez&aacute;visl&eacute; experimenty (n = 3).<\/em><\/p>\n<\/div><\/div>\n\n\n\n<h3 class=\"wp-block-heading\">&Uacute;&#269;inky 5-ALA na infekciu FIPV v ma&#269;acej bunkovej l&iacute;nii<\/h3>\n\n\n\n<p class=\"wp-block-paragraph\">Antiv&iacute;rusov&eacute; &uacute;&#269;inky 5-ALA proti FIPV sa hodnotili testom inhib&iacute;cie plaku v bunk&aacute;ch fcwf-4. Bunky boli lie&#269;en&eacute; 5-ALA nasleduj&uacute;cimi 3 postupmi: 24-hodinov&aacute; predbe&#382;n&aacute; lie&#269;ba (pre-24h), 24-h predbe&#382;n&aacute; lie&#269;ba nasledovan&aacute; 49-hodinovou lie&#269;bou FIPV (pre-24h a co-49h) a 48-hodinov&aacute; predbe&#382;n&aacute; lie&#269;ba, po ktorej nasledovala 49-hodinov&aacute; lie&#269;ba FIPV (pre-48h a co-49h). Pri pre-24h sa percento inhib&iacute;cie plaku v&yacute;znamne zv&yacute;&scaron;ilo pri 500&mu;M alebo vy&scaron;&scaron;ej (obr&aacute;zky 2A, B). Pri pre-24h a co-49h sa percento inhib&iacute;cie plakov v&yacute;znamne zv&yacute;&scaron;ilo pri 125&mu;M alebo vy&scaron;&scaron;ej (obr&aacute;zky 2C, D). Pri pre-48h a co-49h dosiahlo percento inhib&iacute;cie plaku pri FIPV typu I pri 125 &mu;M 5-ALA 75% (obr&aacute;zky 2E, F). Kontroln&yacute; nosi&#269;, SFC, nevykazoval &#382;iadne plakov&eacute; inhibi&#269;n&eacute; &uacute;&#269;inky na FCoV za ak&yacute;chko&#318;vek podmienok. Pod&#318;a titra&#269;n&eacute;ho testu bola produkcia FIPV typu I a typu II v&yacute;znamne zn&iacute;&#382;en&aacute; pri 250 a 500&mu;M 5-ALA (obr&aacute;zok 3).<\/p>\n\n\n\n<div class=\"wp-block-image lm-back\"><figure class=\"aligncenter size-large\"><a href=\"https:\/\/www.fipwarriors.eu\/wp-content\/uploads\/2021\/04\/5ala2.jpg\"><img loading=\"lazy\" decoding=\"async\" width=\"767\" height=\"684\" src=\"https:\/\/www.fipwarriors.eu\/wp-content\/uploads\/2021\/04\/5ala2.jpg\" alt=\"\" class=\"wp-image-6938\" srcset=\"https:\/\/www.fipwarriors.eu\/wp-content\/uploads\/2021\/04\/5ala2.jpg 767w, https:\/\/www.fipwarriors.eu\/wp-content\/uploads\/2021\/04\/5ala2-300x268.jpg 300w, https:\/\/www.fipwarriors.eu\/wp-content\/uploads\/2021\/04\/5ala2-673x600.jpg 673w, https:\/\/www.fipwarriors.eu\/wp-content\/uploads\/2021\/04\/5ala2-13x12.jpg 13w, https:\/\/www.fipwarriors.eu\/wp-content\/uploads\/2021\/04\/5ala2-350x312.jpg 350w, https:\/\/www.fipwarriors.eu\/wp-content\/uploads\/2021\/04\/5ala2-64x57.jpg 64w\" sizes=\"auto, (max-width: 706px) 89vw, (max-width: 767px) 82vw, 740px\"\/><\/a><figcaption><strong>Obr&aacute;zok 2.<\/strong><br>Test inhib&iacute;cie plaku FIPV v bunk&aacute;ch fcwf-4 lie&#269;en&yacute;ch 5-ALA. (<strong>A, B<\/strong>) &Uacute;&#269;inky 24-hodinovej predbe&#382;nej lie&#269;by na antiv&iacute;rusov&uacute; aktivitu 5-ALA. R&yacute;chlos&#357; plakovej inhib&iacute;cie FIPV infikovan&yacute;ch buniek fcwf-4 predbe&#382;ne lie&#269;en&yacute;ch 5-ALA po dobu 24 hod&iacute;n. (<strong>C, D<\/strong>) &Uacute;&#269;inky 24-hodinovej predbe&#382;nej lie&#269;by a 48-hodinovej n&aacute;slednej lie&#269;by na antiv&iacute;rusov&uacute; aktivitu 5-ALA. R&yacute;chlos&#357; plakovej inhib&iacute;cie FIPV infikovan&yacute;ch buniek fcwf-4 predbe&#382;ne lie&#269;en&yacute;ch po&#269;as 24 hod&iacute;n a dodato&#269;ne lie&#269;en&yacute;ch po&#269;as 48 hod&iacute;n s 5-ALA. (<strong>E, F<\/strong>) &Uacute;&#269;inky 48-hodinovej predbe&#382;nej lie&#269;by a 48-hodinovej n&aacute;slednej lie&#269;by na antiv&iacute;rusov&uacute; aktivitu 5-ALA. R&yacute;chlos&#357; plakovej inhib&iacute;cie FIPV-infikovan&yacute;ch buniek fcwf-4 predbe&#382;ne lie&#269;en&yacute;ch po dobu 48 hod&iacute;n a n&aacute;sledne 48 hod&iacute;n s 5-ALA. (<strong>A, C, E<\/strong>) typ I FIPV. (<strong>B, D, F<\/strong>) typ II FIPV. &#268;ierna st&#314;pec: 5-ALA. Biela st&#314;pec: Vehikulum (kontroln&eacute; rozp&uacute;&scaron;&#357;adlo). V&yacute;sledky s&uacute; uveden&eacute; ako priemer &plusmn; SE. &Uacute;daje predstavuj&uacute; &scaron;tyri nez&aacute;visl&eacute; experimenty (n = 4). ** p &lt;0,01 (* p &lt;0,05) vs. vehikulum.<\/figcaption><\/figure><\/div>\n\n\n\n<div class=\"wp-block-image lm-back\"><figure class=\"aligncenter size-large\"><a href=\"https:\/\/www.fipwarriors.eu\/wp-content\/uploads\/2021\/04\/5ala3.jpg\"><img loading=\"lazy\" decoding=\"async\" width=\"767\" height=\"691\" src=\"https:\/\/www.fipwarriors.eu\/wp-content\/uploads\/2021\/04\/5ala3.jpg\" alt=\"\" class=\"wp-image-6940\" srcset=\"https:\/\/www.fipwarriors.eu\/wp-content\/uploads\/2021\/04\/5ala3.jpg 767w, https:\/\/www.fipwarriors.eu\/wp-content\/uploads\/2021\/04\/5ala3-300x270.jpg 300w, https:\/\/www.fipwarriors.eu\/wp-content\/uploads\/2021\/04\/5ala3-666x600.jpg 666w, https:\/\/www.fipwarriors.eu\/wp-content\/uploads\/2021\/04\/5ala3-13x12.jpg 13w, https:\/\/www.fipwarriors.eu\/wp-content\/uploads\/2021\/04\/5ala3-350x315.jpg 350w, https:\/\/www.fipwarriors.eu\/wp-content\/uploads\/2021\/04\/5ala3-64x58.jpg 64w\" sizes=\"auto, (max-width: 706px) 89vw, (max-width: 767px) 82vw, 740px\"\/><\/a><figcaption><strong>Obr&aacute;zok 3.<\/strong><br>Inhib&iacute;cia infekcie FIPV 5-ALA v bunk&aacute;ch fcwf-4. (<strong>A, C<\/strong>) &Uacute;&#269;inky 48-hodinovej predbe&#382;nej lie&#269;by a 48-hodinovej n&aacute;slednej lie&#269;by na antiv&iacute;rusov&uacute; aktivitu 250 &mu;M 5-ALA. (<strong>B, D<\/strong>) &Uacute;&#269;inky 48-hodinovej predbe&#382;nej lie&#269;by a 48-hodinovej n&aacute;slednej lie&#269;by na antiv&iacute;rusov&uacute; aktivitu 500 &mu;M 5-ALA. (<strong>A, B<\/strong>) 250 &mu;M 5-ALA. (<strong>A, B<\/strong>) typu I FIPV. (<strong>C, D<\/strong>) typu II FIPV. &#268;ierny kr&uacute;&#382;ok: 5-ALA. Biely kr&uacute;&#382;ok: vehikulum (kontroln&eacute; rozp&uacute;&scaron;&#357;adlo). V&yacute;sledky s&uacute; uveden&eacute; ako priemer &plusmn; SE. &Uacute;daje predstavuj&uacute; &scaron;tyri nez&aacute;visl&eacute; experimenty (n = 4). ** p &lt;0,01 (* p &lt;0,05) vs. vehikulum.<\/figcaption><\/figure><\/div>\n\n\n\n<h3 class=\"wp-block-heading\">&Uacute;&#269;inky 5-ALA na infekciu FIPV v ma&#269;acej bunkovej l&iacute;nii<\/h3>\n\n\n\n<p class=\"wp-block-paragraph\">Makrof&aacute;gy infikovan&eacute; FIPV sa podie&#318;aj&uacute; na progresii sympt&oacute;mov FIP do &#357;a&#382;k&eacute;ho stavu. Sk&uacute;mali sme, &#269;i 5-ALA inhibuje mno&#382;enie FIPV v makrof&aacute;goch. V tomto experimente sa pou&#382;il FIPV 79-1146 typu II s vysokou schopnos&#357;ou mno&#382;enia v prim&aacute;rnych makrof&aacute;goch ma&#269;iek. Produkcia v&iacute;rusu v FIPV-infikovan&yacute;ch makrof&aacute;goch bola zn&iacute;&#382;en&aacute; pri 250&mu;M 5-ALA u troch zo &scaron;tyroch ma&#269;iek (obr&aacute;zok 4).<\/p>\n\n\n\n<div class=\"wp-block-image lm-back\"><figure class=\"aligncenter size-large\"><a href=\"https:\/\/www.fipwarriors.eu\/wp-content\/uploads\/2021\/04\/5ala4.jpg\"><img loading=\"lazy\" decoding=\"async\" width=\"708\" height=\"245\" src=\"https:\/\/www.fipwarriors.eu\/wp-content\/uploads\/2021\/04\/5ala4.jpg\" alt=\"\" class=\"wp-image-6956\" srcset=\"https:\/\/www.fipwarriors.eu\/wp-content\/uploads\/2021\/04\/5ala4.jpg 708w, https:\/\/www.fipwarriors.eu\/wp-content\/uploads\/2021\/04\/5ala4-300x104.jpg 300w, https:\/\/www.fipwarriors.eu\/wp-content\/uploads\/2021\/04\/5ala4-16x6.jpg 16w, https:\/\/www.fipwarriors.eu\/wp-content\/uploads\/2021\/04\/5ala4-350x121.jpg 350w, https:\/\/www.fipwarriors.eu\/wp-content\/uploads\/2021\/04\/5ala4-64x22.jpg 64w\" sizes=\"auto, (max-width: 708px) 100vw, 708px\"\/><\/a><figcaption><strong>Obr&aacute;zok 4.<\/strong><br>Inhib&iacute;cia infekcie FIPV v makrof&aacute;goch. &Uacute;&#269;inky na antiv&iacute;rusov&uacute; aktivitu 5-ALA (250&mu;M) v prim&aacute;rnych makrof&aacute;goch ma&#269;iek. &#268;ierna st&#314;pec: 5-ALA. Biely st&#314;pec: Vehikulum (kontoln&eacute; rozp&uacute;&scaron;&#357;adlo). &Uacute;daje predstavuj&uacute; tri nez&aacute;visl&eacute; experimenty (n = 3).<\/figcaption><\/figure><\/div>\n\n\n\n<h2 class=\"lm-underscore wp-block-heading\">Diskusia<\/h2>\n\n\n\n<p class=\"wp-block-paragraph\">5-ALA je medziproduktom synt&eacute;zy tetrapyrolu u zvierat, rastl&iacute;n a mikroorganizmov (13&ndash;15). V 80. rokoch bola hl&aacute;sen&aacute; mo&#382;nos&#357; &uacute;&#269;innosti 5-ALA u rastlin&aacute;ch (18), ale bolo &#357;a&#382;k&eacute; vyrobi&#357; 5-ALA v dostato&#269;nom mno&#382;stve pre praktick&eacute; pou&#382;itie, preto&#382;e v mikroorganizmoch sa produkuje iba mal&eacute; mno&#382;stvo. Po tom, &#269;o Nishikawa a kol. zaviedla met&oacute;du masovej produkcie 5-ALA s vyu&#382;it&iacute;m bakt&eacute;ri&iacute; (19), &uacute;&#269;innos&#357; 5-ALA sa potvrdila nielen v po&#318;nohospod&aacute;rstve, ale aj v lek&aacute;rskych a biologick&yacute;ch oblastiach. 5-ALA je lacn&yacute; liek a prakticky sa pou&#382;&iacute;va ako doplnok na zlep&scaron;enie v&yacute;konnosti zvierat a imunitn&uacute; odpove&#271; v oblasti veterin&aacute;rneho lek&aacute;rstva (14, 20).<\/p>\n\n\n\n<p class=\"wp-block-paragraph\">5-ALA inhibovala rast FIPV v bunk&aacute;ch fcwf-4. Bolo zisten&eacute;, &#382;e kovov&eacute; komplexy metabolitu 5-ALA, protoporfyr&iacute;n IX (PpIX), maj&uacute; antiv&iacute;rusov&uacute; aktivitu (21&ndash;23). Kovov&yacute; komplex PpIX, hem, inhibuje mno&#382;enie v&iacute;rusu dengue (21). Na druhej strane, mno&#382;enie v&iacute;rusu Zika nie je hemom inhibovan&eacute; (23). Nie je jasn&eacute;, &#269;i hem inhibuje mno&#382;enie FCoV. Spravidla zv&yacute;&scaron;enie intracelul&aacute;rneho hemu stimuluje produkciu hemeoxygen&aacute;zy-1 (HO-1), enz&yacute;mu degraduj&uacute;ceho hem. Uv&aacute;dza sa, &#382;e HO-1 indukuje antiv&iacute;rusov&uacute; aktivitu (24, 25). Av&scaron;ak v predbe&#382;nom experimente sme potvrdili, &#382;e hladina expresie HO-1 mRNA v bunk&aacute;ch o&scaron;etren&yacute;ch 250&mu;M 5-ALA ostala nezmenen&aacute; (&uacute;daje nie s&uacute; uveden&eacute;). Na z&aacute;klade tohto poznatku doch&aacute;dza pri 5-ALA-indukovanej inhib&iacute;cii FIPV infekcie v d&ocirc;sledku in&eacute;ho faktora ako hem a HO-1.<\/p>\n\n\n\n<p class=\"wp-block-paragraph\">V s&uacute;&#269;asnosti u&#382; existuje ve&#318;a &scaron;t&uacute;di&iacute; o terapeutick&yacute;ch liekoch na FIP. Bolo identifikovan&yacute;ch ve&#318;a liekov &uacute;&#269;inn&yacute;ch na FIP in vitro a bolo potvrden&eacute;, &#382;e niektor&eacute; vykazuj&uacute; lie&#269;ebn&eacute; &uacute;&#269;inky pri pod&aacute;van&iacute; ma&#269;k&aacute;m s FIP (10&ndash;12). &Uacute;&#269;inky v&scaron;etk&yacute;ch liekov v&scaron;ak boli slab&eacute; v pr&iacute;padoch s neurologick&yacute;mi prejavmi. Za pr&iacute;&#269;inu sa pova&#382;oval zl&yacute; prenos t&yacute;chto liekov do centr&aacute;lneho nervov&eacute;ho syst&eacute;mu; preto je potrebn&yacute; liek vykazuj&uacute;ci antiv&iacute;rusov&eacute; &uacute;&#269;inky proti FIPV schopn&eacute;mu dosta&#357; sa do mozgov&eacute;ho tkaniva. 5-ALA je aminokyselina s n&iacute;zkou molekulovou hmotnos&#357;ou a mo&#382;nou dobrou pentera&#269;nou schopnos&#357;ou do mozgov&eacute;ho tkaniva (26). &#270;alej sa uv&aacute;dza, &#382;e dif&uacute;zia 5-ALA z krvi do norm&aacute;lneho mozgov&eacute;ho tkaniva je ve&#318;mi n&iacute;zka (27), &#269;o nazna&#269;uje, &#382;e vykazuje menej nepriazniv&yacute;ch &uacute;&#269;inkov. FIP mo&#382;no jednozna&#269;ne diagnostikova&#357; iba detekciou antig&eacute;nu FCoV v l&eacute;zii (28). Ke&#271; sa v&scaron;ak lie&#269;ba za&#269;ne a&#382; po stanoven&iacute; jednozna&#269;nej diagn&oacute;zy, pr&iacute;znaky medzit&yacute;m progradovali do stavu, ke&#271; u&#382; v mnoh&yacute;ch pr&iacute;padoch sa nedostav&iacute; reakcia na lie&#269;bu. Preto, ak je k dispoz&iacute;cii liek, ktor&yacute; je mo&#382;n&eacute; profylakticky poda&#357; u&#382; pred diagnostikovan&iacute;m FIP, je mo&#382;n&eacute; zabr&aacute;ni&#357; progresii pr&iacute;znakov, pre &#269;o m&ocirc;&#382;e by&#357; 5-ALA ide&aacute;lnym prostriedkom. V na&scaron;om experimente s cie&#318;ov&yacute;mi bunkami FIPV, makrof&aacute;gmi, v&scaron;ak antiv&iacute;rusov&eacute; &uacute;&#269;inky 5-ALA neboli u niektor&yacute;ch ma&#269;iek pozorovan&eacute;. Preto, ke&#271; sa 5-ALA pou&#382;&iacute;va ako terapeutick&eacute; lie&#269;ivo na FIP, anti-FCoV lieky, ako s&uacute; GS-441524 (29), GC-376 (30), U18666A (31) a itrakonazol (32) alebo protiz&aacute;palov&eacute; lieky, ako napr&iacute;klad anti-TNF-alfa <a class=\"glossaryLink\" aria-describedby=\"tt\" data-cmtooltip=\"&lt;div class=glossaryItemBody&gt;&amp;lt;!-- wp:paragraph --&amp;gt;&amp;lt;strong&amp;gt;Protil&aacute;tka&amp;lt;\/strong&amp;gt;&nbsp;(&amp;lt;strong&amp;gt;imunoglobulin&amp;lt;\/strong&amp;gt;) je&nbsp;protein, kter&yacute; je schopen jako sou&#269;&aacute;st&nbsp;imunitn&iacute;ho syst&eacute;mu&nbsp;identifikovat a zne&scaron;kodnit ciz&iacute; objekty (bakterie&nbsp;a&nbsp;viry) v t&#283;le. Protil&aacute;tky jsou nositeli&nbsp;humor&aacute;ln&iacute; imunity. Jsou to krevn&iacute; b&iacute;lkoviny vznikaj&iacute;c&iacute; v m&iacute;zn&iacute; tk&aacute;ni.&amp;lt;br\/&amp;gt;&amp;lt;!-- \/wp:paragraph --&amp;gt;&lt;\/div&gt;\" href=\"https:\/\/www.fipwarriors.eu\/en\/glossary\/protilatka\/\" data-mobile-support=\"0\" data-gt-translate-attributes='[{\"attribute\":\"data-cmtooltip\", \"format\":\"html\"}]' tabindex=\"0\" role=\"link\">protil&aacute;tka<\/a> (33), by mali by&#357; s&uacute;be&#382;ne pou&#382;&iacute;van&eacute; aj &#271;alej.<\/p>\n\n\n\n<p class=\"wp-block-paragraph\">V ter&eacute;ne doch&aacute;dza medzi ma&#269;kami hlavne k prenosu FECV, zatia&#318; &#269;o horizont&aacute;lna infekcia FIPV medzi ma&#269;kami sa pova&#382;uje za zriedkav&uacute; (34). Zistilo sa, &#382;e FIPV je generovan&yacute; genetickou mut&aacute;ciou FECV. Ak teda existuj&uacute; prostriedky na prevenciu infekcie FECV na dennej b&aacute;ze, m&ocirc;&#382;e sa zabr&aacute;ni&#357; rozvoju FIP. Nie je vyvinut&aacute; &#382;iadna vakc&iacute;na schopn&aacute; zabr&aacute;ni&#357; infekcii FECV. Addie a kol. uviedli, &#382;e vylu&#269;ovanie v&iacute;rusov&eacute;ho g&eacute;nu vo v&yacute;kaloch prestalo u ma&#269;iek infikovan&yacute;ch FECV lie&#269;en&yacute;ch syntetick&yacute;m anal&oacute;gom adenoz&iacute;nu (35). Preto sa o&#269;ak&aacute;va elimi&aacute;cia FECV infikuj&uacute;ceho &#269;revo podan&iacute;m ativirotika FECV infikovan&yacute;m ma&#269;k&aacute;m. Syntetick&eacute; anal&oacute;gy adenoz&iacute;nu v&scaron;ak m&ocirc;&#382;u indukova&#357; mut&aacute;ciu g&eacute;nu koronav&iacute;rusu (36). Navy&scaron;e, aj ke&#271; hladina g&eacute;nu FECV vo v&yacute;kaloch klesla pod hranicu detekcie u ma&#269;iek lie&#269;en&yacute;ch syntetick&yacute;m adenoz&iacute;nov&yacute;m anal&oacute;gom, je mo&#382;n&eacute;, &#382;e FECV latentne infikoval &#269;revo alebo in&eacute; tkaniv&aacute;. Aby sa zabr&aacute;nilo mut&aacute;cii g&eacute;nu FECV a spo&#318;ahlivo sa eliminovala infekcia FECV, je potrebn&eacute; dlhodob&eacute; pod&aacute;vanie syntetick&eacute;ho anal&oacute;gu adenoz&iacute;nu, ale to nie je kv&ocirc;li jeho vysokej cene re&aacute;lne. Na druhej strane sa 5-ALA sa prakticky pou&#382;&iacute;va ako doplnok stravy. 5-ALA m&aacute; n&iacute;zku toxicitu pre zvierat&aacute; a rastliny, &#269;o v&yacute;razne nazna&#269;uje mo&#382;nos&#357; dlhodob&eacute;ho pod&aacute;vania 5-ALA ma&#269;k&aacute;m. V bud&uacute;cnosti je potrebn&eacute; presk&uacute;ma&#357;, &#269;i je 5-ALA pou&#382;ite&#318;n&yacute; ako doplnok na zabr&aacute;nenie rozvoja FIP u ma&#269;iek infikovan&yacute;ch FECV.<\/p>\n\n\n\n<p class=\"wp-block-paragraph\">V tejto &scaron;t&uacute;dii sme potvrdili mo&#382;nos&#357;, &#382;e 5-ALA inhibuje mno&#382;enie FIPV a produkciu TNF-alfa. Preto&#382;e 5-ALA je aminokyselina pr&iacute;tomn&aacute; v tele, je mo&#382;n&eacute; jej okam&#382;it&eacute; podanie. Je v&scaron;ak potrebn&eacute; pod&aacute;va&#357; 5-ALA ma&#269;k&aacute;m s FIP a sledova&#357;, &#269;i je mo&#382;n&eacute; dosiahnu&#357; terapeutick&eacute; &uacute;&#269;inky. &#270;alej treba sk&uacute;ma&#357;, &#269;i dlhodob&eacute; pod&aacute;vanie 5-ALA eliminuje v&iacute;rus a inhibuje v&yacute;voj FIP u ma&#269;iek infikovan&yacute;ch FECV.<\/p>\n\n\n\n<h2 class=\"lm-underscore wp-block-heading\">Referencie<\/h2>\n\n\n\n<ol class=\"lm-small wp-block-list\"><li>ICTV&nbsp;<em>Coronaviridae. Virus Taxonomy: 2019 Release<\/em>. (2020). 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The search for other drugs useful in the treatment of FIP continues. 5-aminolevulinic acid is another and certainly not the last candidate.  <\/p>","protected":false},"author":1,"featured_media":6918,"comment_status":"closed","ping_status":"open","sticky":false,"template":"","format":"standard","meta":{"_uag_custom_page_level_css":"","footnotes":""},"categories":[187,190],"tags":[],"class_list":["post-6916","post","type-post","status-publish","format-standard","has-post-thumbnail","hentry","category-odborne","category-preklady"],"featured_image_urls_v2":{"full":["https:\/\/www.fipwarriors.eu\/wp-content\/uploads\/2021\/04\/catlionFHD.jpg",1920,1080,false],"thumbnail":["https:\/\/www.fipwarriors.eu\/wp-content\/uploads\/2021\/04\/catlionFHD-150x150.jpg",150,150,true],"medium":["https:\/\/www.fipwarriors.eu\/wp-content\/uploads\/2021\/04\/catlionFHD-300x169.jpg",300,169,true],"medium_large":["https:\/\/www.fipwarriors.eu\/wp-content\/uploads\/2021\/04\/catlionFHD-768x432.jpg",525,295,true],"large":["https:\/\/www.fipwarriors.eu\/wp-content\/uploads\/2021\/04\/catlionFHD-1024x576.jpg",525,295,true],"1536x1536":["https:\/\/www.fipwarriors.eu\/wp-content\/uploads\/2021\/04\/catlionFHD-1536x864.jpg",1536,864,true],"2048x2048":["https:\/\/www.fipwarriors.eu\/wp-content\/uploads\/2021\/04\/catlionFHD.jpg",1920,1080,false],"trp-custom-language-flag":["https:\/\/www.fipwarriors.eu\/wp-content\/uploads\/2021\/04\/catlionFHD-16x9.jpg",16,9,true],"twentyseventeen-featured-image":["https:\/\/www.fipwarriors.eu\/wp-content\/uploads\/2021\/04\/catlionFHD.jpg",1920,1080,false],"twentyseventeen-thumbnail-avatar":["https:\/\/www.fipwarriors.eu\/wp-content\/uploads\/2021\/04\/catlionFHD-100x100.jpg",100,100,true],"woocommerce_thumbnail":["https:\/\/www.fipwarriors.eu\/wp-content\/uploads\/2021\/04\/catlionFHD-250x250.jpg",250,250,true],"woocommerce_single":["https:\/\/www.fipwarriors.eu\/wp-content\/uploads\/2021\/04\/catlionFHD-350x197.jpg",350,197,true],"woocommerce_gallery_thumbnail":["https:\/\/www.fipwarriors.eu\/wp-content\/uploads\/2021\/04\/catlionFHD-100x100.jpg",100,100,true]},"post_excerpt_stackable_v2":"<p>Tomomi Takano, Kumi Satoh, and Tomoyoshi Doki ? H&#318;adanie &#271;al&scaron;&iacute;ch liekov pou&#382;ite&#318;n&yacute;ch pre lie&#269;bu <a class=\"glossaryLink\" aria-describedby=\"tt\" data-cmtooltip=\"&lt;div class=glossaryItemBody&gt;Infek&#269;n&aacute; peritonit&iacute;da ma&#269;iek (angl. Feline infectious peritonitis, zau&#382;&iacute;van&aacute; skratka FIP) je infek&#269;n&eacute; ochorenie ma&#269;iek sp&ocirc;sobuj&uacute;ce smr&#357;. N&aacute;zov je mierne zav&aacute;dzaj&uacute;ci, preto&#382;e nejde o z&aacute;pal pobru&scaron;nice (&#269;i&#382;e peritonea), ale o z&aacute;pal ciev (vaskulit&iacute;da). Klinick&eacute; pr&iacute;znaky FIP sa v&scaron;ak &#269;asto prejavuj&uacute; vo forme z&aacute;palu pobru&scaron;nice. V&iacute;rus FIP nap&aacute;da v&scaron;ak aj pohrudnicu &ndash; z toho d&ocirc;vodu je niekedy choroba ozna&#269;ovan&aacute; tie&#382; ako Infek&#269;n&aacute; ma&#269;acia polyserozit&iacute;da (Feline infectious polyserositis).&lt;\/div&gt;\" href=\"https:\/\/www.fipwarriors.eu\/en\/glossary\/fip\/\" data-mobile-support=\"0\" data-gt-translate-attributes='[{\"attribute\":\"data-cmtooltip\", \"format\":\"html\"}]' tabindex=\"0\" role=\"link\">FIP<\/a> pokra&#269;uje. Kyselina 5-aminolevulov&aacute; je &#271;al&scaron;&iacute;m a ur&#269;ite nie posledn&yacute;m kandid&aacute;tom.  <\/p>\n","category_list_v2":"<a href=\"https:\/\/www.fipwarriors.eu\/en\/category\/odborne\/\" rel=\"category tag\">Odborn\u00e9<\/a>, <a href=\"https:\/\/www.fipwarriors.eu\/en\/category\/preklady\/\" rel=\"category tag\">Preklady<\/a>","author_info_v2":{"name":"LM","url":"https:\/\/www.fipwarriors.eu\/en\/author\/starman\/"},"comments_num_v2":"0 comments","featured_image_src":"https:\/\/www.fipwarriors.eu\/wp-content\/uploads\/2021\/04\/catlionFHD.jpg","author_info":{"display_name":"L. M.","author_link":"https:\/\/www.fipwarriors.eu\/en\/author\/starman\/"},"yoast_head":"<!-- This site is optimized with the Yoast SEO plugin v28.8 - https:\/\/yoast.com\/product\/yoast-seo-wordpress\/ -->\n<title>Mo\u017en\u00e1 antiv\u00edrusov\u00e1 aktivita kyseliny 5-aminolevulovej pri infekcii v\u00edrusom infek\u010dnej peritonit\u00eddy ma\u010diek - FIP Warriors CZ\/SK \u00ae<\/title>\n<meta name=\"robots\" content=\"index, follow, max-snippet:-1, max-image-preview:large, max-video-preview:-1\" \/>\n<link rel=\"canonical\" href=\"https:\/\/www.fipwarriors.eu\/en\/mozna-antivirusova-aktivita-kyseliny-5-aminolevulovej-pri-infekcii-virusom-infekcnej-peritonitidy-maciek\/\" \/>\n<meta property=\"og:locale\" content=\"en_GB\" \/>\n<meta property=\"og:type\" content=\"article\" \/>\n<meta property=\"og:title\" content=\"Mo\u017en\u00e1 antiv\u00edrusov\u00e1 aktivita kyseliny 5-aminolevulovej pri infekcii v\u00edrusom infek\u010dnej peritonit\u00eddy ma\u010diek - FIP Warriors CZ\/SK \u00ae\" \/>\n<meta property=\"og:description\" content=\"Tomomi Takano, Kumi Satoh, and Tomoyoshi Doki ? H&#318;adanie &#271;al&scaron;&iacute;ch liekov pou&#382;ite&#318;n&yacute;ch pre lie&#269;bu FIP pokra&#269;uje. Kyselina 5-aminolevulov&aacute; je &#271;al&scaron;&iacute;m a ur&#269;ite nie posledn&yacute;m kandid&aacute;tom.\" \/>\n<meta property=\"og:url\" content=\"https:\/\/www.fipwarriors.eu\/en\/mozna-antivirusova-aktivita-kyseliny-5-aminolevulovej-pri-infekcii-virusom-infekcnej-peritonitidy-maciek\/\" \/>\n<meta property=\"og:site_name\" content=\"FIP Warriors CZ\/SK \u00ae\" \/>\n<meta property=\"article:publisher\" content=\"https:\/\/www.facebook.com\/groups\/fipczsk\" \/>\n<meta property=\"article:published_time\" content=\"2021-04-18T07:12:39+00:00\" \/>\n<meta property=\"article:modified_time\" content=\"2023-03-29T07:50:04+00:00\" \/>\n<meta property=\"og:image\" content=\"https:\/\/www.fipwarriors.eu\/wp-content\/uploads\/2021\/04\/catlionFHD.jpg\" \/>\n\t<meta property=\"og:image:width\" content=\"1920\" \/>\n\t<meta property=\"og:image:height\" content=\"1080\" \/>\n\t<meta property=\"og:image:type\" content=\"image\/jpeg\" \/>\n<meta name=\"author\" content=\"L. M.\" \/>\n<meta name=\"twitter:card\" content=\"summary_large_image\" \/>\n<meta name=\"twitter:label1\" content=\"Written by\" \/>\n\t<meta name=\"twitter:data1\" content=\"L. M.\" \/>\n\t<meta name=\"twitter:label2\" content=\"Estimated reading time\" \/>\n\t<meta name=\"twitter:data2\" content=\"17 minutes\" \/>\n<script type=\"application\/ld+json\" class=\"yoast-schema-graph\">{\"@context\":\"https:\\\/\\\/schema.org\",\"@graph\":[{\"@type\":\"Article\",\"@id\":\"https:\\\/\\\/www.fipwarriors.eu\\\/mozna-antivirusova-aktivita-kyseliny-5-aminolevulovej-pri-infekcii-virusom-infekcnej-peritonitidy-maciek\\\/#article\",\"isPartOf\":{\"@id\":\"https:\\\/\\\/www.fipwarriors.eu\\\/mozna-antivirusova-aktivita-kyseliny-5-aminolevulovej-pri-infekcii-virusom-infekcnej-peritonitidy-maciek\\\/\"},\"author\":{\"name\":\"L. 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