Tam Jacqueline Chor Wing, Ko Chun Hay, Koon Chi Man, Cheng Zhang, Lok Wong Hing, Lau Ching Po, Leung Ping Chung, Fung Kwok Pui, Chan Wai Yee, Lau Clara Bik San
Institute of Chinese Medicine, The Chinese University of Hong Kong, Shatin, New Territories, Hong Kong; State Key Laboratory of Phytochemistry and Plant Resources in West China, The Chinese University of Hong Kong, Shatin, New Territories, Hong Kong.
Institute of Chinese Medicine, The Chinese University of Hong Kong, Shatin, New Territories, Hong Kong; State Key Laboratory of Phytochemistry and Plant Resources in West China, The Chinese University of Hong Kong, Shatin, New Territories, Hong Kong; School of Biomedical Sciences, The Chinese University of Hong Kong, Shatin, New Territories, Hong Kong.
PLoS One. 2015 Oct 2;10(10):e0139342. doi: 10.1371/journal.pone.0139342. eCollection 2015.
Angiogenesis is vitally important in diabetic wound healing. We had previously demonstrated that a Chinese 2-herb formula (NF3) significantly stimulated angiogenesis of HUVEC in wound healing. However, the molecular mechanism has not yet been elucidated. In line with this, global expression profiling of NF3-treated HUVEC was performed so as to assess the regulatory role of NF3 involved in the underlying signaling pathways in wound healing angiogenesis. The microarray results illustrated that different panels of differentially expressed genes were strictly governed in NF3-treated HUVEC in a time-regulated manner. The microarray analysis followed by qRT-PCR and western blotting verification of NF3-treated HUVEC at 6 h revealed the involvement of various genes in diverse biological process, e.g., MAP3K14 in anti-inflammation; SLC5A8 in anti-tumorogenesis; DNAJB7 in protein translation; BIRC5, EPCAM, INSL4, MMP8 and NPR3 in cell proliferation; CXCR7, EPCAM, HAND1 and MMP8 in migration; CXCR7, EPCAM and MMP8 in tubular formation; and BIRC5, CXCR7, EPCAM, HAND1, MMP8 and UBD in angiogenesis. After 16 h incubation of NF3, other sets of genes were shown with differential expression in HUVEC, e.g., IL1RAPL2 and NR1H4 in anti-inflammation; miR28 in anti-tumorogenesis; GRIN1 and LCN1 in anti-oxidation; EPB41 in intracellular signal transduction; PRL and TFAP2A in cell proliferation; miR28, PRL and SCG2 in cell migration; PRL in tubular formation; and miR28, NR1H4 and PRL in angiogenesis. This study provided concrete scientific evidence in support of the regulatory role of NF3 on endothelial cells involved in wound healing angiogenesis.
血管生成在糖尿病伤口愈合中至关重要。我们之前已经证明，一种中药两味配方（NF3）在伤口愈合过程中能显著刺激人脐静脉内皮细胞（HUVEC）的血管生成。然而，其分子机制尚未阐明。与此一致的是，对经NF3处理的HUVEC进行了全基因组表达谱分析，以评估NF3在伤口愈合血管生成的潜在信号通路中所起的调控作用。微阵列结果表明，在经NF3处理的HUVEC中，不同组的差异表达基因以时间调控的方式受到严格控制。对经NF3处理6小时的HUVEC进行微阵列分析，随后进行qRT-PCR和蛋白质印迹验证，结果显示多种基因参与了不同的生物学过程，例如，MAP3K14参与抗炎；SLC5A8参与抗肿瘤发生；DNAJB7参与蛋白质翻译；BIRC5、EPCAM、INSL4、MMP8和NPR3参与细胞增殖；CXCR7、EPCAM、HAND1和MMP8参与迁移；CXCR7、EPCAM和MMP8参与管状结构形成；BIRC5、CXCR7、EPCAM、HAND1、MMP8和UBD参与血管生成。在NF3孵育16小时后，HUVEC中显示出其他几组差异表达的基因，例如，IL1RAPL2和NR1H4参与抗炎；miR28参与抗肿瘤发生；GRIN1和LCN1参与抗氧化；EPB41参与细胞内信号转导；PRL和TFAP2A参与细胞增殖；miR28、PRL和SCG2参与细胞迁移；PRL参与管状结构形成；miR28、NR1H4和PRL参与血管生成。本研究为NF3对参与伤口愈合血管生成的内皮细胞的调控作用提供了具体的科学证据。
