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培养的大鼠血管平滑肌细胞中神经营养因子及其受体的基因表达

Gene expression of neurotrophins and their receptors in cultured rat vascular smooth muscle cells.

作者信息

Nemoto K, Fukamachi K, Nemoto F, Miyata S, Hamada M, Nakamura Y, Senba E, Ueyama T

机构信息

Laboratory of Health Science, School of Pharmaceutical Sciences, University of Shizuoka, 52-l Yada, Shizuoka, 422-8526, Japan.

出版信息

Biochem Biophys Res Commun. 1998 Apr 7;245(1):284-8. doi: 10.1006/bbrc.1998.8418.

Abstract

Most previous researches on neurotrophins including nerve growth factor (NGF), brain-derived neurotrophic factor (BDNF), and neurotrophin-3 (NT-3) have focused on the nervous system, because their receptors are widely distributed in neuronal tissues. Recently, however, the participation of neurotrophins in inflammation and atherosclerosis has been proposed. Therefore, the gene expression of neurotrophins is now an urgent issue is to be investigated in nonneuronal tissues. Here, we evaluated the gene expression of neurotrophins and their receptors in rat cultured vascular smooth muscle cells (VSMCs) by the reverse transcriptase-polymerase chain reaction method. The transcripts of NGF, NT-3, and TrkC (high-affinity receptor for NT-3), and two BDNF alternative spliced transcript variants with exons 3 and 4 were clearly detected in VSMCs cultured under conventional culture conditions. The upregulation of mRNA levels for NGF, two BDNF variants with exons 1 and 2, low-affinity neurotrophin receptor, and high-affinity receptors, TrkA (for NGF) and TrkB (for BDNF), was observed in response to the treatment with serum and phorbol-ester following the serum-starvation. In contrast, the expression of NT-3 and TrkC genes was downregulated under these conditions. Co-expression of these factors and their receptors and the characteristic regulation of their gene transcriptions suggest that these factors play crucial roles in the function of VSMCs through an autocrine mechanism.

摘要

此前,大多数关于神经营养因子(包括神经生长因子(NGF)、脑源性神经营养因子(BDNF)和神经营养因子-3(NT-3))的研究都集中在神经系统,因为它们的受体广泛分布于神经组织中。然而,最近有人提出神经营养因子参与了炎症和动脉粥样硬化过程。因此,神经营养因子在非神经组织中的基因表达是目前亟待研究的问题。在此,我们采用逆转录-聚合酶链反应法评估了大鼠培养血管平滑肌细胞(VSMC)中神经营养因子及其受体的基因表达。在常规培养条件下培养的VSMC中,可清晰检测到NGF、NT-3和TrkC(NT-3的高亲和力受体)的转录本,以及两个带有外显子3和4的BDNF可变剪接转录变体。血清饥饿后,用血清和佛波酯处理可使NGF、两个带有外显子1和2的BDNF变体、低亲和力神经营养因子受体以及高亲和力受体TrkA(针对NGF)和TrkB(针对BDNF)的mRNA水平上调。相反,在这些条件下,NT-3和TrkC基因的表达下调。这些因子及其受体的共表达以及它们基因转录的特征性调节表明,这些因子通过自分泌机制在VSMC的功能中发挥关键作用。

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