本文關(guān)鍵詞：hsa-miR-143在人脂肪細(xì)胞誘導(dǎo)分化過程中的表達變化及調(diào)控因素分析，由筆耕文化傳播整理發(fā)布。

        目的（1）探討hsa-miR-143在脂肪細(xì)胞分化前后的表達變化規(guī)律；（2）對hsa-miR-143進行系統(tǒng)的生物信息學(xué)分析，預(yù)測hsa-miR-143的靶基因和可能參與的功能；（3）FFAs（free fatty acids，游離脂肪酸）、TNF-（tumor necrosisfactor-alpha，腫瘤壞死因子alpha）、IL-6（interleukin6，白介素6）、leptin（瘦素）、resistin（抵抗素）分別對人成熟脂肪細(xì)胞hsa-miR-143表達的影響。方法（1）利用Real time PCR技術(shù)，檢測脂肪細(xì)胞分化前后hsa-miR-143的表達水平；（2）應(yīng)用miRbase、NCBI Mapviewer、UCSC Genome Browser等在線工具獲取并分析hsa-miR-143的序列特征；應(yīng)用TargetScan，PicTar及miRanda預(yù)測hsa-miR-143的靶基因，取三個預(yù)測結(jié)果的交集，并合并DIANALAB-TarBase6.0數(shù)據(jù)庫的已驗證靶標(biāo)，對所得靶基因集分別進行功能富集分析（Gene Ontology, GO）和信號轉(zhuǎn)導(dǎo)通路富集分析（Pathway Enrichment）；應(yīng)用pubmed、google等信息搜索工具綜述hsa-miR-143已有功能研究；（3）利用Realtime PCR技術(shù)檢測FFAs、TNF-、IL-6、leptin、resistin干預(yù)成熟脂肪細(xì)胞前后hsa-miR-143的表達水平的變化。結(jié)果（1）分化成熟的人脂肪細(xì)胞hsa-miR-143的表達量較分化前顯著上調(diào)（P<0.05）；（2）hsa-miR-143在各物種之間具有高度保守性，功能注釋顯示預(yù)測靶基因主要富集于肌動蛋白細(xì)胞骨架組織合成、Rac蛋白信號轉(zhuǎn)導(dǎo)的正調(diào)、突觸傳遞的調(diào)控等生物學(xué)過程（P<0.001）；（3）Pathway分析顯示預(yù)測靶基因顯著富集于肌動蛋白細(xì)胞骨架調(diào)節(jié)、黑色素瘤、細(xì)胞縫隙連接、MAPK（Mitogen-activated protein kinase，絲裂原激活蛋白激酶）信號通路等（P<0.05）；（4）FFAs, leptin及resistin顯著下調(diào)人成熟脂肪細(xì)胞hsa-miR-143的表達（P<0.05），但TNF-及IL-6對其表達量影響較小（P>0.05）。結(jié)論（1）hsa-miR-143在成熟人脂肪細(xì)胞中，其表達量達到相對較高的水平，說明在脂肪細(xì)胞分化成熟過程中，其有一定的調(diào)控作用，生物信息學(xué)分析提示hsa-miR-143可能與細(xì)胞分化、代謝密切相關(guān)；（3）FFAs、leptin及resistin能顯著下調(diào)成熟人脂肪細(xì)胞hsa-miR-143的表達，TNF-及IL-6對成熟人脂肪細(xì)胞hsa-miR-143的表達無明顯作用。研究結(jié)果提示，脂肪細(xì)胞hsa-miR-143的表達，可能受FFAs、leptin及resistin因素的調(diào)節(jié)，并推論，，F(xiàn)FAs、leptin及resistin對miR-143的表達可能存在有一個負(fù)反饋調(diào)節(jié)機制，這一推論仍需驗證。本研究闡明了hsa-miR-143在人脂肪前體細(xì)胞誘導(dǎo)分化過程中的表達規(guī)律，提示了FFAs、Leptin、Resistin等因素對成熟人脂肪細(xì)胞hsa-miR-143的表達具有一定的調(diào)控作用。研究結(jié)果為深入分析hsa-miR-143與肥胖、胰島素敏感性之間的關(guān)系提供了新的線索，并為肥胖的早期干預(yù)及肥胖相關(guān)并發(fā)癥的防治提供新的靶點。

    Objective （1） The different expression of hsa-miR-143in the process ofdifferentiation in human adipocytes was investigated.（2）To predict target gene andfunctions of hsa-miR-143by bioinformatic analysis.（3） The alteration ofhsa-miR-143expression in matured human adipocytes were studied after intervenedby FFAs and some adipokines such as TNF-, IL-6, leptin and resistin.Methods The levels of hsa-miR-143expression in the human preadipocytes thathad differentiated into mature adipocytes were analyzed using quantitative real timePCR. MiRBase, NCBI Mapviewer, UCSC Genome Browser database were used toget the sequence of hsa-miR-143. The target gene were predicted by TargetScan,PicTar and miRanda, and the intersection of results of three database with knowntarget gene were analyzed by Gene Ontology and pathway analysis. Pubmed andgoogle were used to review previous studies of hsa-miR-143. And furthermore, theexpression of hsa-miR-143were measured in mature adipocytes treated with FFAs,TNF-, IL-6, leptin and resistin were analyzed using quantitative real time PCR.Results （1）During the conversion of cultured human preadipocytes to matureadipocytes, the expression of hsa-miR-143was upregulated (P<0.05).（2）Hsa-miR-143was highly conserved among species. By gene ontology analysis, thetarget genes were enriched in actin cytoskeleton organization and biogenesis,positively regulating Rac protein signal transduction, synaptic transmission, and otherbiological processes (P<0.001).（3）By pathway analysis, the target genes weremainly involved in regulation of actin cytoskeleton, Melanoma, Gap junction, MAPKsignaling pathway, and others (P<0.05).（4）The expression of hsa-miR-143were downregulated by FFAs, leptin and resistin in human adipocytes (P<0.05), but not byTNF-and IL-6(P>0.05).Conclusions （1）During the conversion of cultured human preadipocytes to matureadipocytes, the expression of hsa-miR-143is upregulated. Bioinformatic Analysisresults show hsa-miR-143is closely related to cell differentiation and metabolism.（2）hsa-miR-143is inhibited by FFAs,leptin, and resistin,but not by TNF-and IL-6in cultered human adipocytes. These findings indicate that FFAs, leptin, and resistin,may inhibit the expression of miR-143via a negative feedback which requires furtherstudy. In conclusion, we identify the regularity of miR-143expression during theconversion of human preadipocytes into mature adipocytes and several regulativefactors on the miR-143expression in cultured human adipocytes. These data providefurther evidence of the partial involvement of hsa-miR-143in the regulation ofinsulin sensitivity mediated by certain adipokines. Our findings may bring newinsights into the mechanisms of obesity-associated insulin resistance.

hsa-miR-143在人脂肪細(xì)胞誘導(dǎo)分化過程中的表達變化及調(diào)控因素分析

中文摘要5-7Abstract7-8前言9-12第一部分 hsa-miR-143 在人脂肪細(xì)胞誘導(dǎo)分化過程中的表達變化及生物信息學(xué)分析12-37    材料與方法12-23    結(jié)果23-34    討論34-37第二部分 hsa-miR-143 在成熟人脂肪細(xì)胞中調(diào)控因素的分析37-49    材料與方法37-40    結(jié)果40-46    討論46-49參考文獻49-55中英文縮略語55-56綜述56-65    參考文獻62-65已撰寫和發(fā)表文章65-66致謝66

  本文關(guān)鍵詞：hsa-miR-143在人脂肪細(xì)胞誘導(dǎo)分化過程中的表達變化及調(diào)控因素分析，由筆耕文化傳播整理發(fā)布。