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Identification of Target Genes Involved in Wound Healing Angiogenesis of Endothelial Cells with the Treatment of a Chinese 2-Herb Formula.

Tam JC, Ko CH, Koon CM, Cheng Z, Lok WH, Lau CP, Leung PC, Fung KP, Chan WY, Lau CB - PLoS ONE (2015)

Bottom Line: We had previously demonstrated that a Chinese 2-herb formula (NF3) significantly stimulated angiogenesis of HUVEC in wound healing.The microarray results illustrated that different panels of differentially expressed genes were strictly governed in NF3-treated HUVEC in a time-regulated manner.This study provided concrete scientific evidence in support of the regulatory role of NF3 on endothelial cells involved in wound healing angiogenesis.

View Article: PubMed Central - PubMed

Affiliation: 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.

ABSTRACT
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.

No MeSH data available.


Related in: MedlinePlus

Western blotting of NF3 on HUVEC for 6 and 16 h.Immunobloting was performed three times using independent cell lysates and representative blots of (A) 6 h and (B) 16 h were shown.
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pone.0139342.g004: Western blotting of NF3 on HUVEC for 6 and 16 h.Immunobloting was performed three times using independent cell lysates and representative blots of (A) 6 h and (B) 16 h were shown.

Mentions: Western blotting of the protein expression was conducted so as to confirm the differentially expressed gene targets revealed from microarray data. Since a panel of different sets of genes was believed to be involved in HUVEC after NF3 treatment for 6 and 16 h, two representative proteins at each time point were selected for further confirmation. The selection criteria of the proteins from these sets of genes were based on the role of the genes and the availability of antibody for western blotting. CXCR7 and MMP8 were used at 6 h while NR1H4 and SCG2 were examined at 16 h. As shown in Fig 4, NF3 caused a strong induction of the protein expression of CXCR7 in HUVEC associated with an increase in the protein expression of MMP8 and NR1H4 in HUVEC. A mild increase in the protein expression of SCG2 was observed in NF3-treated HUVEC. The results demonstrated that both genes and proteins were tightly involved and regulated in HUVEC after NF3 treatment, promoting the overall wound healing angiogenesis.


Identification of Target Genes Involved in Wound Healing Angiogenesis of Endothelial Cells with the Treatment of a Chinese 2-Herb Formula.

Tam JC, Ko CH, Koon CM, Cheng Z, Lok WH, Lau CP, Leung PC, Fung KP, Chan WY, Lau CB - PLoS ONE (2015)

Western blotting of NF3 on HUVEC for 6 and 16 h.Immunobloting was performed three times using independent cell lysates and representative blots of (A) 6 h and (B) 16 h were shown.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4591983&req=5

pone.0139342.g004: Western blotting of NF3 on HUVEC for 6 and 16 h.Immunobloting was performed three times using independent cell lysates and representative blots of (A) 6 h and (B) 16 h were shown.
Mentions: Western blotting of the protein expression was conducted so as to confirm the differentially expressed gene targets revealed from microarray data. Since a panel of different sets of genes was believed to be involved in HUVEC after NF3 treatment for 6 and 16 h, two representative proteins at each time point were selected for further confirmation. The selection criteria of the proteins from these sets of genes were based on the role of the genes and the availability of antibody for western blotting. CXCR7 and MMP8 were used at 6 h while NR1H4 and SCG2 were examined at 16 h. As shown in Fig 4, NF3 caused a strong induction of the protein expression of CXCR7 in HUVEC associated with an increase in the protein expression of MMP8 and NR1H4 in HUVEC. A mild increase in the protein expression of SCG2 was observed in NF3-treated HUVEC. The results demonstrated that both genes and proteins were tightly involved and regulated in HUVEC after NF3 treatment, promoting the overall wound healing angiogenesis.

Bottom Line: We had previously demonstrated that a Chinese 2-herb formula (NF3) significantly stimulated angiogenesis of HUVEC in wound healing.The microarray results illustrated that different panels of differentially expressed genes were strictly governed in NF3-treated HUVEC in a time-regulated manner.This study provided concrete scientific evidence in support of the regulatory role of NF3 on endothelial cells involved in wound healing angiogenesis.

View Article: PubMed Central - PubMed

Affiliation: 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.

ABSTRACT
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.

No MeSH data available.


Related in: MedlinePlus