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Effect of Yifei Huoxue Granule (益肺活血颗粒) on the proliferation of rat pulmonary artery smooth muscle cells upon exposure to chronic hypoxic conditions in vitro

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Abstract

Objective

To investigate the inhibitory effect of Yifei Huoxue Granule (益肺活血颗粒, YFHXG): on the hypoxia-induced proliferation of rat pulmonary artery smooth muscle cells (PASMCs) and its mechanism of decreasing pulmonary arterial pressure.

Methods

Twenty male Sprague-Dawley (SD) rats were randomly: divided into four groups: saline, and 0.66, 3.30 and 16.50 g/kg of YFHXG groups, the saline and different concentrations of YFHXG were given twice daily for 7 days, respectively. Serum-pharmacology method was used in the preparation of YFHXG serum. Tissue block anchorage was employed in the primary culture of rat PASMCs. The PASMCs were randomly divided into normoxia group, hypoxia group, and hypoxia+YFHXG group (0.66, 3.30 and 16.50 g/kg doses of YFHXG-treated serum groups, exposed to hypoxic condition). PASMCs in normoxia and hypoxia group were cultured with saline serum, hypoxia+YFHXG groups were cultured with different concentrations of YFHXG serum. Cell viability was assessed with 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. Cell cycle was analyzed using flow cytometry. In addition, hypoxia inducible factor-1-alpha (HIF-1α) protein expression was evaluated by immunocytochemistry analysis, the concentration of intracellular reactive oxygen species (ROS) and Ca Ca2+ were determined by laser scanning confocal microscopy (LSCM).

Results

MTT assay and flow cytometry showed that hypoxia could directly activate the proliferation of PASMCs, while YFHXG dose-dependently inhibited hypoxia-induced proliferation of rat PASMCs. Immunocytochemistry showed that hypoxia enhanced HIF-1α protein expression, and LSCM showed that hypoxia significantly increased intracellular ROS and Ca Ca2+, while YFHXG decreased the expression of HIF-, 1α and attenuated the hypoxia-induced increase in intracellular concentration of ROS and Ca Ca2+.

Conclusions

YFHXG could inhibit hypoxia-induced proliferation of rat PASMCs, which may decrease pulmonary arterial pressure and vascular remodeling. The anti-hypoxia effect of YFHXG may be explained by its regulation of HIF-1α expression and of the levels of intracellular ROS and Ca Ca2+.

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Authors and Affiliations

  1. Institute of Very Important Person Pulmonary Medicine, Navy General Hospital of People’s Liberation Army, Beijing, 100048, China

    Ling-yun Zhang  (张凌云) & Min Ou  (欧 敏)

  2. Institute of Hematology, Navy General Hospital of People’s Liberation Army, Beijing, 100048, China

    You-zhang Huang  (黄友章)

  3. Institute of Central Laboratory, Navy General Hospital of People’s Liberation Army, Beijing, 100048, China

    Yuan-yuan Qiao  (乔媛媛) & Da-jin Zhang  (张达矜)

Authors
  1. Ling-yun Zhang  (张凌云)
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  2. Min Ou  (欧 敏)
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  3. You-zhang Huang  (黄友章)
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  4. Yuan-yuan Qiao  (乔媛媛)
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  5. Da-jin Zhang  (张达矜)
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Correspondence to Min Ou  (欧 敏).

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Supported by Military Medical and Health Foundation (No. 36040)

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Zhang, Ly., Ou, M., Huang, Yz. et al. Effect of Yifei Huoxue Granule (益肺活血颗粒) on the proliferation of rat pulmonary artery smooth muscle cells upon exposure to chronic hypoxic conditions in vitro . Chin. J. Integr. Med. 18, 507–513 (2012). https://doi.org/10.1007/s11655-012-1150-7

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  • DOI: https://doi.org/10.1007/s11655-012-1150-7

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