靶向调控ITAM/ITIM-FcγRs及SYK信号级联在糖尿病肾病早期炎症机制中的作用

批准号:
81570651
项目类别:
面上项目
资助金额:
57.0 万元
负责人:
柳飞
依托单位:
学科分类:
H0504.继发性肾脏疾病
结题年份:
2019
批准年份:
2015
项目状态:
已结题
项目参与者:
顾志敏、徐缓、唐江涛、薄虹、张君龙、王婷立、周姣姣、苏晓凤
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中文摘要
炎症机制在早期糖尿病肾病(DN)中起重要作用。以CRP为代表的因子通过与免疫球蛋白G Fc受体(FcγRs)相结合而促进炎症。我们前期研究发现:糖尿病CRP转基因小鼠肾脏表达含免疫受体酪氨酸激活基序的FcγRs(ITAM-FcγRs)上调;高糖刺激系膜细胞ITAM-FcγRs增多,含免疫受体酪氨酸抑制基序的FcγR(ITIM-FcγR)减少,脾酪氨酸激酶(SYK)活化。由此我们假设:ITAM/ITIM-FcγRs表达失衡影响SYK信号级联是DN早期炎症机制的核心。因此,本课题采用体内外转染方法,构建ITAM-FcγRs、SYK基因抑制、ITIM-FcγR过表达的细胞和SD大鼠,高糖刺激细胞和STZ诱导动物模型,在不同时间点检测ITAM/ITIM-FcγRs表达及对SYK信号级联的影响,从分子-细胞-整体水平探讨调控ITAM/ITIM-FcγRs影响SYK信号级联在早期DN的作用机制。
英文摘要
Diabetic nephropathy(DN) is a low grade inflammatory disease trigged by metabolic disorder. Inflammation has an important role in the early stage of DN pathogenesis. Inflammatory cytokines, such as C-reactive protein(CRP),have effects on cell through binding immunoglobulin G Fc receptors (FcγRs) in the surface of cell. We previously found that the expression of FcγRs with an immunoreceptor tyrosine-based activation motif(ITAM-FcγRs) increased in the kidney of diabetic CRP-Tg mouse induced by STZ. ITAM-FcγRs increased, FcγRs with an immunoreceptor tyrosine-based inhition motif(ITIM-FcγR) decreased and spleen tyrosine kinase(SYK) activated in cultured rat glomerular mesangial cells(GMC) ambient high glucose. A role for activated SYK signal cascade leading to the cell responses and pro-inflammatory cytokines production has been established in antibody-dependent kidney disease. Based on previous findings, we hypothesized that the unbalance of ITAM /ITIM-FcγRs and activation of SYK play a central role of inflammatory mechanism in the early stage of DN. Therefore, we plan to construct the GMC, tubular epithelial cells and SD rats models of ITAM- FcγRs , SYK gene inhibition and ITIM-FcγR overpxression(wild-type)by transfection in vivo and vitro in the current study. Then, the cells will be stimulated by high glucose and the rats treated with streptozotocin. The expression and activity of ITAM-FcγRs, ITIM-FcγR and SYK gene influences on inflammatory signaling pathways and molecular markers related with fibrosis will be tested at different time points. We aim to investigate the role of ITAM/ITIM-FcγRs unbalance and their effects on SYK cascade in the early stage of DN and to identify new target for treatment and prevention of the disease.
糖尿病肾病(diabetic nephropathy, DN)是糖尿病最主要的微血管并发症,在世界范围内都是终末期肾病(end stage renal disease, ESRD)的首位病因。其发病机制至今尚未完全阐明,缺乏有效的干预措施。探讨DN的发病机制和早期防治具有重大的社会意义和经济价值。既往研究发现炎症机制在早期糖尿病肾病(DN)中起重要作用。我们前期研究发现:糖尿病CRP转基因小鼠肾脏表达含免疫受体酪氨酸激活基序的FcγRs(ITAM-FcγRs)上调;高糖刺激系膜细胞ITAM-FcγRs增多,含免疫受体酪氨酸抑制基序的FcγR(ITIM-FcγR)减少,脾酪氨酸激酶(SYK)活化。本课题在前期研究基础上采用基因重组技术,构建SYKshRNA的质粒,分别进行体外转染人肾小管上皮细胞株HK-2和体内转染C57BL6糖尿病小鼠模型,在不同时间点,检测ITAM/ITIM-FcγRs表达及对SYK及其信号级联-核转录因子-κB (NF-κB)、下游信号通路关键炎症因子白介素-1β(IL-1β)、肿瘤坏死因子-α(TNF-α)表达的影响,分析肾组织病理变化。我们的研究发现DN成模小鼠肾组织炎症细胞浸润明显,尿蛋白增加,ITAM-FcγRs表达上调,SYK表达及磷酸化增加,NF-κB信号通路激活,炎症因子IL-1β、TNF-α表达上调。予以SYKshRNA干预后,DN小鼠肾组织炎症细胞浸润明显减轻,尿蛋白减少,部分抑制激活的NF-κB信号通路,降低炎症因子IL-1β、TNF-α表达,证实ITAM/ITIM-FcγRs表达失衡激活SYK信号级联是DN早期炎症机制的核心,为靶向调控SYK这一“信号开关”分子防治DN早期炎症损伤,延缓DN肾小球硬化及肾小管间质纤维化奠定理论基础。
期刊论文列表
专著列表
科研奖励列表
会议论文列表
专利列表
The activation of immunoglobulin G Fc receptors (Fc gamma Rs) with immunoreceptor tyrosine-based activation motifs (ITAMs) promotes cognitive impairment in aged rats with diabetes
免疫球蛋白 G Fc 受体 (Fc gamma Rs) 与基于免疫受体酪氨酸的激活基序 (ITAM) 的激活可促进老年糖尿病大鼠的认知障碍
DOI:10.1016/j.exger.2019.110660
发表时间:2019
期刊:Experimental Gerontology
影响因子:3.9
作者:Yue Qiang;Meng Zhangmin;Wang Lingxiao;Sun Qianqian;Wang Shuang;Li Jun;Liu Fei
通讯作者:Liu Fei
国内基金
海外基金
