目的通过串联质谱标签(tandem mass tag.TMT)定量蛋白质组学分析胸主动脉瘤患者及动脉瘤基础上并发StanfordA型主动脉夹层的患者,与正常人群主动脉管壁的差异表达蛋白,筛选出与胸主动脉瘤/夹层中膜退行性病变密切相关的蛋白标志物。方...目的通过串联质谱标签(tandem mass tag.TMT)定量蛋白质组学分析胸主动脉瘤患者及动脉瘤基础上并发StanfordA型主动脉夹层的患者,与正常人群主动脉管壁的差异表达蛋白,筛选出与胸主动脉瘤/夹层中膜退行性病变密切相关的蛋白标志物。方法收集2019年12月至2020年5月四川大学华西医院心脏移植捐献者[正常组,年龄(40.50±9.31)岁]、升主动脉瘤患者[动脉瘤组,年龄(56.50±8.19)岁]及动脉瘤基础上并发Stanford A型主动脉夹层患者[夹层组,年龄(54.17±6.68)岁]的升主动脉组织样本各6例,均为男性。采用TMT.定量蛋白质谱技术,筛选出动脉瘤组和夹层组与正常组之间具有显著表达差异(差异倍数≥1.50或≤0.67)的蛋白质,通过GO和KEGG功能富集分析探讨潜在的信号通路机制。结果相比正常组,动脉瘤组和夹层组均有表达且具有显著差异的蛋白质共63个,其中发生上调的蛋白质有30个,下调的蛋白质有33个。基于GO富集分析及KEGG通路分析,筛选出动脉瘤及夹层组的3条共同差异代谢通路,分别为过氧化物酶体增殖物激活受体(PPAR)信号通路.细胞外基质受体相互作用(extracellular matrix receptor interaction)信号通路以及补体和凝血级联(complement and coagulation cascades)信号通路。结论本研究筛选了在胸主动脉瘤/夹层发生发展过程中具有潜在重要作用的63个差异蛋白质及3条差异代谢通路.为搜寻调控胸主动脉瘤/夹层中膜退行性病变过程中的关键分子靶标提供了依据。展开更多
This study investigated the effects of SIRT1 gene knock-out on osteoarthritis in mice, and the possible roles of SREBP2 protein and the PI3K/AKT signaling pathway in the effects. Mice were randomly divided into a norm...This study investigated the effects of SIRT1 gene knock-out on osteoarthritis in mice, and the possible roles of SREBP2 protein and the PI3K/AKT signaling pathway in the effects. Mice were randomly divided into a normal group and a SIRT1 gene knock-out group（6 mice in each group）. In these groups, one side of the knee anterior cruciate ligament was traversed, and the ipsilateral medial meniscus was cut to establish an osteoarthritis model of knee joint. The countralateral synovial bursa was cut out, serving as controls. The knee joint specimens were then divided into four groups： SIRT1^（＋/＋） control group（group A, n=6）; SIRT1^（＋/＋） osteoarthritis group（group B, n=6）; SIRT1^（–/–） control group（group C, n=6）; SIRT1^（–/–） osteoarthritis group（group D, n=6）. HE staining, Masson staining, Safranin O-Fast Green staining and Van Gieson staining were used to observe the morphological changes in the articular cartilage of the knee. Immunohistochemical staining was employed to detect the expression of SIRT1, SREBP2, VEGF, AKT, HMGCR and type Ⅱ collagen proteins. SA-β-gal staining was utilized to evaluate chondrocyte aging. The results showed clear knee joint cartilage destruction and degeneration in the SIRT1^（–/–） osteoarthritis group. The tidal line was twisted and displaced anteriorly. Type Ⅱ collagen was destroyed and distributed unevenly. Compared with the SIRT1^（＋/＋） osteoarthritis group and SIRT1^（–/–） control group, SIRT1 protein expression was not obviously changed in the SIRT1^（–/–） osteoarthritis group（P〉0.05）, while the expression levels of the SREBP2, VEGF and HMGCR proteins were significantly increased（P〈0.05） and the levels of AKT and type Ⅱ collagen proteins were significantly decreased（P〈0.05）. SIRT1 gene knock-out may aggravate cartilage degeneration in osteoarthritis by activating the SREBP2 protein-mediated PI3K/AKT signalling pathway, suggesting that SIRT1 gene may play a protective role again展开更多
文摘目的通过串联质谱标签(tandem mass tag.TMT)定量蛋白质组学分析胸主动脉瘤患者及动脉瘤基础上并发StanfordA型主动脉夹层的患者,与正常人群主动脉管壁的差异表达蛋白,筛选出与胸主动脉瘤/夹层中膜退行性病变密切相关的蛋白标志物。方法收集2019年12月至2020年5月四川大学华西医院心脏移植捐献者[正常组,年龄(40.50±9.31)岁]、升主动脉瘤患者[动脉瘤组,年龄(56.50±8.19)岁]及动脉瘤基础上并发Stanford A型主动脉夹层患者[夹层组,年龄(54.17±6.68)岁]的升主动脉组织样本各6例,均为男性。采用TMT.定量蛋白质谱技术,筛选出动脉瘤组和夹层组与正常组之间具有显著表达差异(差异倍数≥1.50或≤0.67)的蛋白质,通过GO和KEGG功能富集分析探讨潜在的信号通路机制。结果相比正常组,动脉瘤组和夹层组均有表达且具有显著差异的蛋白质共63个,其中发生上调的蛋白质有30个,下调的蛋白质有33个。基于GO富集分析及KEGG通路分析,筛选出动脉瘤及夹层组的3条共同差异代谢通路,分别为过氧化物酶体增殖物激活受体(PPAR)信号通路.细胞外基质受体相互作用(extracellular matrix receptor interaction)信号通路以及补体和凝血级联(complement and coagulation cascades)信号通路。结论本研究筛选了在胸主动脉瘤/夹层发生发展过程中具有潜在重要作用的63个差异蛋白质及3条差异代谢通路.为搜寻调控胸主动脉瘤/夹层中膜退行性病变过程中的关键分子靶标提供了依据。
基金supported by grants from the National Natural Science Foundation of China(No.81272032)the Research and Development Projects of Shenzhen of China(Nos.JCYJ20150403091443275)
文摘This study investigated the effects of SIRT1 gene knock-out on osteoarthritis in mice, and the possible roles of SREBP2 protein and the PI3K/AKT signaling pathway in the effects. Mice were randomly divided into a normal group and a SIRT1 gene knock-out group（6 mice in each group）. In these groups, one side of the knee anterior cruciate ligament was traversed, and the ipsilateral medial meniscus was cut to establish an osteoarthritis model of knee joint. The countralateral synovial bursa was cut out, serving as controls. The knee joint specimens were then divided into four groups： SIRT1^（＋/＋） control group（group A, n=6）; SIRT1^（＋/＋） osteoarthritis group（group B, n=6）; SIRT1^（–/–） control group（group C, n=6）; SIRT1^（–/–） osteoarthritis group（group D, n=6）. HE staining, Masson staining, Safranin O-Fast Green staining and Van Gieson staining were used to observe the morphological changes in the articular cartilage of the knee. Immunohistochemical staining was employed to detect the expression of SIRT1, SREBP2, VEGF, AKT, HMGCR and type Ⅱ collagen proteins. SA-β-gal staining was utilized to evaluate chondrocyte aging. The results showed clear knee joint cartilage destruction and degeneration in the SIRT1^（–/–） osteoarthritis group. The tidal line was twisted and displaced anteriorly. Type Ⅱ collagen was destroyed and distributed unevenly. Compared with the SIRT1^（＋/＋） osteoarthritis group and SIRT1^（–/–） control group, SIRT1 protein expression was not obviously changed in the SIRT1^（–/–） osteoarthritis group（P〉0.05）, while the expression levels of the SREBP2, VEGF and HMGCR proteins were significantly increased（P〈0.05） and the levels of AKT and type Ⅱ collagen proteins were significantly decreased（P〈0.05）. SIRT1 gene knock-out may aggravate cartilage degeneration in osteoarthritis by activating the SREBP2 protein-mediated PI3K/AKT signalling pathway, suggesting that SIRT1 gene may play a protective role again
