Effects of hypoxia on the expression of HMGB1 and related inflammatory factors in human pulmonary artery smooth muscle and pulmonary artery endothelial cells

doi:10.12047/j.cjap.5746.2019.088

Abstract

Abstract: Objective: To investigate the effects of hypoxia on the expressions of high mobility group box-1(HMGB1), HMGB1 receptors and inflammatory factors in human pulmonary artery smooth muscle cell( HPASMC) and human pulmonary artery endothelial cells (HPAEC).The effects of HMGB1 on the proliferation and migration activity of the two kinds of cells were detected. Methods: HPASMC and HPAEC were cultured under hypoxic conditions (Hypoxia at 1% oxygen, Hypoxia group) and normoxic conditions(Control group) . The mRNA levels of HMGB1, Toll-like receptors 2,4,9 (TLR2 , TLR4, TLR9), the receptor of advanced glycation end products(RAGE) , CD24 and IL-6 ,TNF-α,CXCL8 were detected by real-time PCR. Cell proliferation was measured by MTS. Cell migration was analysed by wound healing test. Results: Compared with control group, the expressions of HMGB1 mRNA and RAGE mRNA in both HPASMC and HPAEC were increased significantly (P＜0.05 and 0.01). Meanwhile, the expressions of CD24 mRNA in HPAEC and IL-6 mRNA in HPASMC of hypoxia group were increased significantly (P＜0.05). MTS results showed that HMGB1 inhibited the proliferation of HPAEC at 345 pmol/L significantly (P＜0.01). HMGB1 had no effect on the proliferation of HPASMC. Wound healing test showed that HMGB1 had no significant effect on HPASMC and HPAEC. Conclusion: HMGB1 was produced by HPAEC and HPASMC induced by hypoxia. HMGB1 induces endothelial barrier dysfunction by inhibiting HPAEC proliferation. Hypoxia stimulates HPASMC to produce inflammatory cytokines.

Key words: hypoxia, high-mobility group box 1, human pulmonary artery endothelial cells, human pulmonary artery smooth muscle cells, rat

BU Bao-ying, XU Xi-yuan, HAN Jun-ping, YANG Jing-ping. Effects of hypoxia on the expression of HMGB1 and related inflammatory factors in human pulmonary artery smooth muscle and pulmonary artery endothelial cells[J]. CJAP, 2019, 35(5): 406-409.

References

[1] 武敏, 杨敬平, 徐喜媛, 等. 人质子感知受体G2A和OGR1在低氧性肺动脉高压患者外周血细胞中的表达[J]. 中国应用生理学杂志, 2017, 33(1): 85-88.
[2] Harris HE, Andersson U, Pisetsky DS. HMGB1: a multifunctional alarmin driving autoimmune and inflammatory disease[J]. Nat Rev Rheumatol, 2012, 8(4): 195-202.
[3] Sadamura-Takenaka Y, Ito T, Noma S, et al. HMGB1 promotes the development of pulmonary arterial hypertension in rats[J]. PLoS One, 2014, 9(7): e102482.
[4] Humbert M, Sitbon O, Chaouat A, et al. Survival in patients with idiopathic, familial, and anorexigen-associated pulmonary arterial hypertension in the modern management era[J]. Circulation, 2010, 122(2): 156－163.
[5] Humbert M, Morrell NW, Archer SL, et al. Cellular and molecular pathobiology of pulmonary arterial hypertension[J]. J Am Coll Cardiol, 2004, 43(12 Suppl S): 13S－24S.
[6] Ferhani N, Letuve S, Kozhich A, et al. Expression of high-mobility group box 1 and of receptor for advanced glycation and products in chronic obstructive pulmonary disease[J]. Am J Respir Crit Care Med, 2010, 181(9): 917－927.
[7] Chen YS, Wang XJ, Feng WW, et al. Advanced glycation end products decrease collagen I levels in fibroblasts from the vaginal wall of patients with POP via the RAGE, MAPK and NF-κB pathways[J]. Int J Mol Med, 2017, 40(4): 987－998.
[8] Lin S, Man L, Xun M, et al. Inhibition of HMGB1 reduces rat spinal cord astrocytic swelling and AQP4 expression after oxygen-glucose deprivation and reoxygenation via TLR4 and NF-κB signaling in an IL-6-dependent manner[J]. J Neuro Inflammation, 2017, 14(1): 231.
[9] Tikellis C, Thomas MC, Harcourt BE, et al. Cardiac inflammation associated with a Western diet is mediated via activation of RAGE by AGEs[J]. Am J Physiol Endocrinol Metab, 2008, 295(2): E323－330.
[10]Moser B, Megerle A, Bekos C, et al. Local and systemic RAGE axis changes in pulmonary hypertension: CTEPH and iPAH[J]. PLoS One, 2014, 9(9): e106440.
[11]Sadamura-Takenaka Y, Ito T, Noma S, et al. HMGB1 promotes the development of pulmonary arterial hypertension in rats[J]. PLoS One, 2014, 9(7): e102482.
[12]Savale L, Tu L, Rideau D, et al. Impact of interleukin-6 on hypoxia-induced pulmonary hypertension and lung inflammation in mice[J]. Respir Res, 2009, 10(1): 6.
[13]Wang HL, Peng LP, Chen WJ, et al. HMGB1 enhances smooth muscle cell proliferation and migration in pulmonary artery remodeling[J]. Int J Clin Exp Pathol, 2014, 7(7): 3836-3844.
[14]Diana Z, Slaven C, Xu H, et al. High-mobility group box-1 induces vascular remodelling processes via c-Jun activation[J]. J Cell Mol Med, 2015,19(5):1151-1161.
[15]Bauer EM, Shapiro R, Billiar TR, et al. High mobility group Box 1 inhibits human pulmonary artery endothelial cell migration via a Toll-like receptor 4- and interferon response factor 3-dependent mechanism(s)[J]. J Biol Chem, 2013, 288(2): 1365－1373.
[16]Hayakawa K, Pham LD, Katusic ZS, et al. Astrocytic high-mobility group box 1 promotes endothelial progenitor cell-mediated neurovascular remodeling during stroke recovery[J]. Proc Natl Acad Sci U S A, 2012, 109(19): 7505－7510.
[17]Degryse B, Bonaldi T, Scaffidi P, et al. The high mobility group (HMG) boxes of the nuclear protein HMG1 induce chemotaxis and cytoskeleton Reorganization in rat smooth muscle cells[J]. J Cell Biol, 2001, 152(6): 1197－1206.