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      1. 當前位置:首頁 > 智慧健康列表 > 細胞分子生物學 > 詳細信息
        N-WASP 磷酸化在肺炎衣原體感染促進血管新生中的作用
        來源: | 作者: | 發布時間:2013-11-8 15:54:37

        N-WASP 磷酸化在肺炎衣原體感染促進血管新生中的作用
        王海偉,王蓓蓓,張利軍,馬路,劉靜雅,張麗莙
        基金項目:國家自然科學基金資助項目(No.30971225、No.81300206);高等學校博士點基金項目(No.
        20111202110011)
        作者簡介:王海偉(1986 年-),女,研究生,主要研究方向:動脈粥樣硬化
        通信聯系人:張麗莙(1963 年-),女,教授,主要研究方向:動脈粥樣硬化. 

         (天津醫科大學基礎醫學院生理學與病理生理學系,天津300070)
        摘要:目的 探討N-WASP 磷酸化在肺炎衣原體(C.pn)感染誘導血管新生中的作用及其可能機制。方法 C.pn 增殖培養后感染人血管內皮細胞(VEC),免疫熒光染色確認感染成功;Western blot 檢測C.pn 感染的VEC 內N-WASP 的磷酸化水平;CCK-8 檢測N-WASP 特異性抑制劑Wiskostain 對VEC 活力的影響;免疫熒光實驗檢測工作濃度的Wiskostain 在使用時間內對C.pn 感染率的影響;C.pn 感染Wiskostain(5 μM)預處理的VEC 后,Western blot檢測N-WASP 的磷酸化水平,Capillary tube formation 實驗觀察各組VEC 形成新生血管能力的變化。結果 在熒光顯微鏡下,感染的VEC 胞漿內可見典型的C.pn 包涵體。C.pn 感染VEC10 h 和24 h 后N-WASP 的磷酸化水平均明顯上調且高于正常對照組;經Wiskostain 預處理后,C.pn 感染上調N-WASP 磷酸化水平的作用則明顯被抑制(P < 0.05)。Capillary tube formation 實驗結果顯示,C.pn 感染VEC 16 h 后,其所形成的微管腔節點數明顯多于對照組(P < 0.05);經Wiskostain 預處理后,C.pn 感染促進微管腔節點形成的作用則被顯著削弱,幾乎不能形成微管腔結構(P < 0.05)。結論 C.pn 感染可能通過誘導N-WASP 磷酸化促進VEC 形成新生血管。
        關鍵詞:肺炎衣原體;血管新生;N-WASP;動脈粥樣硬化
        中圖分類號:R363
        Roles of N-WASP phosphorylation in angiogenesis promoted
        by Chlamydia pneumoniae infection
        WANG Haiwei, WANG Beibei, ZHANG Lijun, MA Lu, LIU Jingya, ZHANG Lijun
         (Department of Physiology and Pathophysiology, School of Basic Medical Sciences, Tianjin Medical University, Tianjin 300070, China)
        Abstract: Objective To explore the roles of N-WASP phosphorylation in Chlamydia pneumoniae(C.pn) infection-induced angiogenesis and the possible mechanisms in this process. Method VECs were infected with C.pn after its propagation. The successful infection of C.pn with VECs was identified by immumofluorescence. Western blot was performed to determine N-WASPphosphorylation level. The cytotoxicity of Wiskostain was assessed by a CCK-8 assay. The effects of Wiskostain (5 μM) on C.pn infection of VECs were determined by immumofluorescence. After VECs were pretreated with Wiskostain (5 μM), Western blot was used to detect the changes in N-WASP phosphorylation levels in VECs from each group, and capillary tube formation assay was performed to observe the changes in the abilities of infected VECs to form new blood vessels.
        Results The typical C.pn inclusions were observed in the cytoplasm of the infected VECs under a fluorescence microscope. Western blot results showed that the phosphorylation levels of N-WASP were signigicantly increased 10 h and 24 h after infection, and were higher than that in control VECs, which was significantly inhibited when VECs were pretreated with Wiskostain (P < 0.05).The results from capillary tube formation assay showed that the number of network branch points

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