Motoshima Hiroyuki, Wu Xiangdong, Mahadev Kalyankar, Goldstein Barry J
Dorrance Hamilton Research Laboratories, Division of Endocrinology, Diabetes and Metabolic Diseases, Department of Medicine, Jefferson Medical College of Thomas Jefferson University, Philadelphia, PA 19107, USA.
Biochem Biophys Res Commun. 2004 Mar 5;315(2):264-71. doi: 10.1016/j.bbrc.2004.01.049.
Adiponectin (also known as 30-kDa adipocyte complement-related protein or Acrp30) is an abundant adipocyte-derived plasma protein with anti-atherosclerotic and insulin-sensitizing properties. In order to investigate the potential mechanism(s) of the vascular protective effect of adiponectin, we used cultured bovine endothelial cells (BAECs) to study the effect of recombinant globular adiponectin (gAd) on cellular proliferation and the generation of reactive oxygen species (ROS) induced by oxidized LDL (oxLDL). By RT-PCR, we found that BAECs preferentially express AdipoR1, the high-affinity receptor for gAd. Treatment of BAECs with oxLDL (10 microg/ml) for 16h stimulated cell proliferation by approximately 60%, which was inhibited by co-incubation with gAd. Cell treatment with gAd also inhibited basal and oxLDL-induced superoxide release, and suppressed the activation of p42/p44 MAP kinase by oxLDL. The effects of gAd were blocked by a specific polyclonal anti-adiponectin antibody (TJ414). OxLDL-induced BAEC proliferation and superoxide release were inhibited by the NAD(P)H oxidase inhibitor diphenyleneiodonium (DPI), but not the eNOS inhibitor l-nitroarginine methyl ester (l-NAME). Finally, gAd ameliorated the suppression of eNOS activity by oxLDL. These data indicate that gAd inhibits oxLDL-induced cell proliferation and suppresses cellular superoxide generation, possibly through an NAD(P)H oxidase-linked mechanism.
脂联素（也称为30 kDa脂肪细胞补体相关蛋白或Acrp30）是一种丰富的源自脂肪细胞的血浆蛋白，具有抗动脉粥样硬化和胰岛素增敏特性。为了研究脂联素血管保护作用的潜在机制，我们使用培养的牛内皮细胞（BAECs）来研究重组球状脂联素（gAd）对细胞增殖以及氧化型低密度脂蛋白（oxLDL）诱导的活性氧（ROS）生成的影响。通过逆转录聚合酶链反应（RT-PCR），我们发现BAECs优先表达gAd的高亲和力受体AdipoR1。用oxLDL（10微克/毫升）处理BAECs 16小时可使细胞增殖刺激约60%，而与gAd共同孵育可抑制这种增殖。用gAd处理细胞还可抑制基础状态和oxLDL诱导的超氧化物释放，并抑制oxLDL对p42/p44丝裂原活化蛋白激酶（MAP激酶）的激活。gAd的作用被特异性多克隆抗脂联素抗体（TJ414）阻断。oxLDL诱导的BAEC增殖和超氧化物释放被烟酰胺腺嘌呤二核苷酸磷酸（NAD(P)H）氧化酶抑制剂二苯基碘鎓（DPI）抑制，但不被内皮型一氧化氮合酶（eNOS）抑制剂L-硝基精氨酸甲酯（L-NAME）抑制。最后，gAd改善了oxLDL对eNOS活性的抑制作用。这些数据表明，gAd可能通过NAD(P)H氧化酶相关机制抑制oxLDL诱导的细胞增殖并抑制细胞超氧化物生成。
