胰岛素促进大鼠体外血管平滑肌细胞 RANKL表达.docVIP

  胰岛素促进大鼠体外血管平滑肌细胞 RANKL 表达# 周华1,3，袁凌青1，朱佳花2，汪化文2，刘幼硕2，廖二元1** 5 10 15 20 25 30 35 40 （1. 中南大学湘雅二医院代谢内分泌研究所； 2. 中南大学湘雅二医院老年科； 3. 郴州市第一人民医院内分泌科） 摘要：目的：探讨胰岛素对大鼠体外钙化型血管平滑肌细胞（CVSMCs） RANKL 表达的 影响及其细胞信号机制。方法：取不同浓度及时间点，用胰岛素干预大鼠 CVSMCs。应用 RT-PCR 和 ELISA 方法分别测定 CVSMCs RANKL mRNA 和 RANKL 蛋白在大鼠 CVSMCs 的表达。胰岛素以及联合 ERK1/2 信号转导阻断剂 PI3K 信号转导阻断剂和 AKt 信号转导阻 断剂干预大鼠 CVSMCs，利用 Western Blot 检测 p-ERK1/2，ERK1/2，p-Akt，Akt 的表达情 况。胰岛素以及联合 ERK1/2 信号转导阻断剂 PD98509、PI3K 信号转导阻断剂 LY294002 和 AKt 信号转导阻断剂 HIMO 干预大鼠 CVSMCs，利用 RT-PCR 技术检测 RANKL mRNA 表 达水平。结果：1.胰岛素促进 CVSMCs RANKL mRNA 和 RANKL 蛋白的表达。2.胰岛素呈 时间依赖性促进 CVSMCs RANKL mRNA 表达。3.胰岛素诱导大鼠 CVSMCs ERK1/2 以及 Akt 磷酸化。4.ERK1/2 信号转导阻断剂 PD98509 干预组抑制胰岛素促进大鼠 CVSMCs RANKL mRNA 表达增加效应，而 PI3K 信号转导阻断剂 LY294002 组和 AKt 信号转导阻断 剂 HIMO 组无该抑制效应。结论：胰岛素通过激活 ERK1/2 信号转导途径促进大鼠 CVSMCs RANKL 表达。 关键词：胰岛素；钙化型血管平滑肌细胞；RANKL the effects of insulin on the expression of RANKL in rat Vascular smooth muscle cells Zhou Hua1, Yuan Lingqing1, Zhu Jiahua2, Wang Huawen2, Liu Youshuo2, Liao Eryuan3 (1. Institute of Endocrinology and Metabolism, Second Xiangya Hospital of Central South University; 2. Geriatric Department, The Second Xiang-Ya Hospital, Central South University; 3. Institute of Endocrinology and Metabolism, Second Xiangya Xiang-Ya Hospital, Central South University) Abstract: Objective，To investigate the effects and the signal pathway invovled in insulin-induced RANKL expression in rat CVSMCs. Methods：The expression of RANKL mRNA in CVSMCs was detected by RT-PCR and RANKL protein was analyzed by ELISA. Insulin, PD98059 (an inhibitor of ERK 1/2), LY294002 (an inhibitor of PI3-K) or HIMO (an inhibitor of Akt) treated CVSMCs in vitro, p-ERK1/2, ERK1/2, p-Akt, Akt were analysed by Western blot. The expression of RANKL mRNA in rat VSMCs was detected by RT-PCR. Results: 1.Insulin increased the expression of RANKL mRNA and protein in rat CVSMCs .2. Insulin increased the expression of RANKL mRNA in a time-dependent manner. 3.Insulin induced ERK 1/2 and Akt signal pathway in r