Final publication is available at http://dx.doi.org/10.1016/j.exer.2012.08.013. Congrong Guo, Atsushi Otani, Akio Oishi, Hiroshi Kojima, Yukiko Makiyama, Satoko Nakagawa, Nagahisa Yoshimura, Knockout of ccr2 alleviates photoreceptor cell death in a model of retinitis pigmentosa, Experimental Eye Research, Volume 104, November 2012, Pages 39-47, ISSN 0014-4835 / 京都大学 / 0048 / 新制・課程博士 / 博士(医学) / 甲第18680号 / 医博第3952号 / 新制||医||1007(附属図書館) / 31613 / 京都大学大学院医学研究科医学専攻 / (主査)教授 長澤 丘司, 教授 伊藤 壽一, 教授 長田 重一 / 学位規則第4条第1項該当 / Doctor of Medical Science / Kyoto University / DFAM

Adipocytokines released by the adipose tissue play an important role in the regulation of immune and inflammatory responses. In obesity their production is dysregulated, which is one of the major factors contributing to the onset of a chronic low-grade systemic inflammation representing a risk factor for the progression of other diseases, such as atherosclerosis or type-2 diabetes. The main goal of this thesis was to analyze the secretion of selected adipocytokines (adiponectin, IL6 and MCP1) by in-vitro differentiated adipocytes, isolated from the adipose tissue prior to and after a dietary intervention, and this under basal conditions and during stimulated lipolysis. In case of adiponectin, the secretion of its isoforms was analyzed too. The concentration of adiponectin, IL6 and MCP1 was determined by the ELISA method, the Western Blot method was used to determine the distribution of the adiponectin isoforms. The thesis also concentrates on the gene expression of ATF3, ATF4 and HSPA5, factors engaged in the ER stress in the course of the differentiation of adipocytes. The changes in the gene expression were measured by the quantitative Real Time PCR method. At the same time the development of the endoplasmic reticulum (ER) in the course of adipogenesis was monitored by indirect...

<p>U okviru ove doktorske disertacije ispitan je hemijski sastav i biolo&scaron;ke aktivnosti metanolnih i vodenih ekstrakata odabranih samoniklih vrsta tribusa Urticeae, rod&nbsp; Urtica:&nbsp; U. <em>dioica</em>&nbsp; subsp.&nbsp; <em>dioica</em>&nbsp; var.&nbsp;<em> pubescens</em>, U.<em>&nbsp; dioica&nbsp;</em> subsp.&nbsp; <em>dioica&nbsp; </em>var.&nbsp; <em>dioica</em>&nbsp; i&nbsp; U. <em>kioviensis</em>&nbsp; i tribusa Parietarieae, rod&nbsp; Parietaria:<em>&nbsp; P. officinalis</em>,&nbsp; <em>P.</em><br /><em>lusitanica</em> L. subsp<em>. lusitanica</em>,<em> P. judaica</em> L. subsp. <em>judaica</em> i<em> P. serbica</em>. Cilj rada bio je da se odredi sadrţaj biolo&scaron;ki aktivnih jedinjenja u ovim, do sada veoma malo ispitanim vrstama famijije Urticaceae, i utvrdi njihov potencijal primene kao pomoćnih lekovitih sredstava i dodataka ishrani.<br />Hemijski sastav ekstrakata ispitivanih vrsta određen je primenom: tečnohromatografskih tehnika (LC-DAD-MS i LC-MS-MS) za kvalitativnu analizu metanolnih ekstrakata, dok je za kvantitativnu analizu odabranih fenolnih jedinjenja primenjena LC-MS-MS tehnika. Spektrofotometrijskim metodama je određen sadržaj<br />ukupnih fenolnih komponenti i flavonoida. Ispitivanja biolo&scaron;kih aktivnosti ekstrakata obuhvatila su: određivanje antioksidantne i antiinflamatorne aktivnosti kao i sposobnost ekstrakata da inhibiraju acetilholinesterazu.&nbsp; Određen je uticaj odabranih metanolnih ekstrakata na imuni odgovor i proliferaciju intestinalnih ćelijskih linija pacova (IEC18) i ĉoveka (Caco2).<br />Dobijeni rezultati ukazuju da odabrane vrste tribusa Urticeae i Parietarieae, odnosno rodova&nbsp; Urtica&nbsp; i Parietaria&nbsp; predstavljaju bogate izvore biolo&scaron;ki aktivnih jedinjenja koja ispoljavaju raznovrsne biolo&scaron;ke aktivnosti. Sa hemotaksonomskog aspekta izdvajaju se sledeća jedinjenja kao potencijalni taksonomski markeri: vi&scaron;i sadržaj 5-O-kafeoilhinske kiseline u ekstraktima herbi vrsta roda&nbsp; Urtica, i visok sadrţaj<br />epikatehina u ekstraktima korena vrsta roda&nbsp; Parietaria. Ekstrakt herbe vrste&nbsp; U. kioviensis&nbsp; se od ostalih izdvaja po tome &scaron;to ne sadrži rutin a sadrži&nbsp; C-glikozide, u najvećoj meri viteksin. Od svih ispitivanih ekstrakata, ekstrakti korena&nbsp; Parietaria&nbsp; vrsta su ispoljili najbolji antioksidantni potencijal u većini izvr&scaron;enih testova. Najsnažniji antiinflamatorni potencijal je ispoljio ekstrakt korena vrste&nbsp; P. officinalis&nbsp; a prate ga ekstrakti korena vrsta roda&nbsp; Urtica.&nbsp; Veoma dobar antiinflamatorni potencijal su ispoljili infuzi herbi vrste U.<em> dioica</em>&nbsp; (čajevi od koprive). Svi ispitani metanolni ekstrakti su ispoljili odličnu inhibiciju enzima acetilholinesteraze a kao najbolji se izdvajaju ekstrakti korena&nbsp; Parietaria&nbsp; vrsta i vrste&nbsp; U.<em> kioviensis</em>. Povećanu sekreciju citokina rat MCP1 i GRO&alpha;&nbsp; izazivaju ekstrakti korena vrsta&nbsp; P. officinalis&nbsp; i&nbsp; P. judaica&nbsp; u<br />bazalnim uslovima i uslovima LPS-stimulisane inlamacije, dok ekstrakti vrste&nbsp; U. dioica&nbsp; povećavaju bazalnu a smanjuju LPS-stimulisanu sekreciju. Stimulaciju sekrecije ova dva citokina, ispitivani ekstrakti vr&scaron;e interakcijom sa adapternim proteinom MyD88 (ali ne intereaguju sa TLR4 receptorom) i NF -&kappa;B signalnim<br />putem. Ekstrakt korena vrste&nbsp; P. <em>officinalis</em>&nbsp; povećava LPS-om indukovanu ekspresiju enzima COX-2 u IEC18 ćelijama, dok je ekstrakt korena vrste&nbsp; U.<em> dioica</em>&nbsp; smanjuje. Efekat epitelizacije ili zarastanja rane na monosloju IEC18 ćelija ispoljavaju ekstrakti herbe i korena vrste&nbsp; P. <em>officinalis</em>. Ispitivani ekstrakti ne menjaju značajno seksreciju citokina hMCP1 i IL-8 u Caco2 ćelijama niti ispoljavaju značajan uticaj na &nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp; njihovu proliferaciju.</p> / <p>Within this doctoral thesis the chemical composition and biological activity of methanol and aqueous extracts of the selected plant species belonging to the Urticeae and Parietarieae tribe, more specifically to the&nbsp; Urtica&nbsp; and&nbsp; Parietaria&nbsp; genuses was evaluated (Urtica:&nbsp; U.<em> dioica</em>&nbsp; subsp.<em> dioica</em>&nbsp; var.&nbsp; <em>pubescens</em>,&nbsp; U.&nbsp; <em>dioica</em>&nbsp; subsp.&nbsp;<em> dioica</em>&nbsp; var.&nbsp; <em>dioica</em>&nbsp; and&nbsp; U.<em> kioviensis</em>;&nbsp; Parietaria:&nbsp; P<em>. officinalis</em>,&nbsp; P.<em> lusitanica</em>&nbsp; subsp.&nbsp; <em>lusitanica,</em>&nbsp; P. <em>judaica&nbsp;</em> subsp.<em>&nbsp; judaica&nbsp;</em> and<em>&nbsp; P. serbica</em>). The principal aim was to determine the content of biologically active&nbsp; compounds in this, poorly<br />examined species of the Urticaceae family, and determine their potential as additional remedy and dietary supplements.<br />Qualitative analysis of methanol extracts was performed by LC-DAD-MS i LC-MS/MS analysis, and LC-MS/MS for quantitative analysis of selected phenolic compounds. Total phenolics and flavonoids were determined spectrophotometrically. In order to assess the biological potential, the antioxidant and anti-inflammatory activities of the extracts were studied as well as&nbsp; their ability to inhibit acetylcholinesterase. The immuno-modulatory effects of the selected methanol extract on the immune response and proliferation of intestinal epithelial cells (IEC18 and Caco2)<br />was determined.<br />The obtained results suggest that the examined species of the Urticeae and Parietarieae tribe (genuses&nbsp; Urtica&nbsp; and&nbsp; Parietaria) are abundant with the biologically active compounds that express a broad spectrum of biological activities. As a potential chemotaxonomic markers stand out the following&nbsp; compounds: 5-O-caffeoilquinic acid (highly abundant in the herb extracts of the&nbsp; Urtica&nbsp; spp.) and epicatechin (highly abundant in the root extracts of the&nbsp; Parietaria&nbsp; spp.).&nbsp; U.<br />kioviensis&nbsp; herb extracts differs from the rest by high content of vitexin and total lack of rutin. The best antioxidant potential have exhibited the root extracts of the&nbsp; Parietaria&nbsp; species. The strongest anti-inflammatory potential had the root extract of the&nbsp; P. officinalis, followed by root extracts of the&nbsp; Urtica&nbsp; spp. Excellent anti-inflammatory activity have exhibited the aqueous&nbsp; extracts of&nbsp; U.<em> dioica</em>&nbsp; herbs&nbsp; &ndash;&nbsp; stinging nettle teas. All tested methanol extracts have inhibited enzyme acetylcholinesterase, the best inhibitors being root extracts of&nbsp; U.<em> kioviensis&nbsp;</em> and Parietaria&nbsp; species. Root extracts of&nbsp; P<em>. officinalis</em>&nbsp; and&nbsp; P.<em> judaica</em>&nbsp; have increased the basal and LPS-stimulated secretion of rat MCP1 and GRO&alpha;, while&nbsp; U. <em>dioica&nbsp;</em> extracts increased the basal but decreased the LPS-stimulated secretion. The examined extracts interact with the MyD88 (but not the TLR4) and NF-&kappa;B signaling pathway. The root extract of&nbsp; P<em>. officinalis&nbsp; </em>increase LPS-stimulated expression of COX-2 in IEC18 cells, while the root extract of&nbsp; U<em>. dioica</em>&nbsp; decreases it.<br />The herb and root extract of P. <em>officinalis</em>&nbsp; exhibit the wound healing effect. Investigated extracts do not significantly alter the secretion of hMCP1 and IL-8 in Caco2 cells and exhibit no significant effect to their proliferation.</p> / null