Higenamine exerts an antispasmodic effect on cold-induced vasoconstriction by regulating the PI3K/Akt, ROS/α2C-AR and PTK9 pathways independently of the AMPK/eNOS/NO axis
Higenamine exerts an antispasmodic effect on cold-induced vasoconstriction by regulating the PI3K/Akt, ROS/α2C-AR and PTK9 pathways independently of the AMPK/eNOS/NO axis
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Higenamine 通过独立于 AMPK/eNOS/NO 轴调节 PI3K/Akt、ROS/α2C-AR 和 PTK9 通路，对寒冷引起的血管收缩发挥解痉作用
DOI：
10.3892/etm.2019.7656
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发表时间：
2019-08-01
影响因子：
2.7
通讯作者：
Zhao, Guoping
中科院分区：
文献类型：
--
作者：
Guan, Jianhua;Lin, Haoming;Zhao, Guoping
本研究旨在探讨去甲乌药碱对冷诱导的皮肤血管收缩的抗痉挛作用及其分子机制。建立冷诱导大鼠皮肤血管收缩模型，静脉注射不同剂量去甲乌药碱。分析组间皮肤局部血流量(RBF)的变化。采用四甲基偶氮唑盐比色法测定体外培养的人真皮微血管内皮细胞增殖情况。硝酸还原酶法测定NO浓度。采用流式细胞仪检测细胞内活性氧自由基(ROS)水平。Western blotting检测蛋白质表达水平。结果表明，模型组大鼠的RBF较正常对照组下降，但去甲乌药碱可逆转模型组大鼠的RBF。内皮型一氧化氮合酶(ENOS)、磷酸化(P)-eNOS、蛋白激酶B(Akt1)、p-Akt1、AMP活化蛋白激酶(AMPK)α1和p-AMPKα1的表达经低温处理后上调，而去甲乌药碱可逆转其表达。去甲乌药碱能显著降低细胞内α2C肾上腺素受体(AR)水平，与亚低温组比较差异有统计学意义(P
The present study aimed to investigate the antispasmodic effect of higenamine on cold-induced cutaneous vasoconstriction and the underlying molecular mechanisms. A cold-induced cutaneous vasoconstriction rat model was established and different doses of higenamine were delivered by intravenous injection. The changes of cutaneous regional blood flow (RBF) between groups were analyzed. In vitro, the proliferation of human dermal microvascular endothelial cells was measured by MTT. The NO concentration was detected by a nitrate reductase assay. Flow cytometry was applied to measure reactive oxygen species (ROS) levels. The protein expression levels were detected by western blotting. The results demonstrated that in the model group, RBF declined compared with the normal control group, but was reversed by treatment with higenamine. The expression of endothelial nitric oxide synthase (eNOS), phosphorylated (p)-eNOS, protein kinase B (Akt1), p-Akt1, AMP-activated protein kinase (AMPK) alpha 1 and p-AMPK alpha 1 was upregulated by hypothermic treatment but was reversed by higenamine treatment. Treatment with higenamine significantly reduced the level of intracellular alpha 2C-adrenoreceptor (AR) compared with the hypothermia group (P