Hypoxia-inducible factor 1α regulates the expression of CD18 and ninjurin-1_Chinese Journal of Ocular Fundus Diseases

Authors：

 SongHuping , ZhuQi , WuQiong , 艾华 , 雷哓琴 , 朱昭亮 , 杨阳

Department of Ophthalmology, Xian NO. 4 Hospital, Xian 710004, China;

Corresponding author：

SongHuping, Email: songhpxian@163.com

Keywords：

Diabetic retinopathy/pathophsiology; Hypoxia-inducible factor 1, alpha subunit; RNA, small interfering

DOI：

10.3760/cma.j.issn.1005-1015.2014.02.010

Video：

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Objective To investigate the effects of hypoxia-inducible factor 1α (HIF-1α) small interfere RNA construct pSUPERH1-siHIF1α on the expression of CD18 and ninjurin-1 by K562 (human chronic myelogenous leukemia cell line) cells cultured with serums from patients with early stage of diabetic retinopathy. Methods K562 cells were cultured in 4 groups as control group (group A), diabetic group (group B), diabetes and pSUPERH1-siHIF1α transfect group (group C) and diabetes and pSUPER-retro transfect group (group D). The cells in group A were cultured in human serum from age-matched healthy control, and in group B, C and D, the cells were cultured in serum from the subjects of early stage of diabetic retinopathy. Twenty-four hours before the cells were cultured by the serum from the subjects of early stage of diabetic retinopathy, the HIF-1α specific siRNA expression vector pSUPERH1-siHIF1α and empty vector pSUPER-retro were transfected into the cells of group C and D, respectively. The percentages of CD18 and ninjurin-1 positive cell on the surface of K562 cells were measured by Flow Cytometry. The adherent rate between K562 and RF/6A was measured by the rose Bengal staining test. Results The percentages of CD18 positive cell in the group A, B, C and D were significantly different (F=14.33, P=0.01). The percentage of group B was significantly higher than that in group A (P=0.001); the percentage of group C was significantly lower than that in group B (P=0.001) and group D (P=0.02); the difference between group C and A was not significant (95%CI=-14.89-2.13, P=0.12). The differences of the percentage of ninjurin-1 positive cell among the group A, B, C and D were significant (F=39.38, P=0.001). The percentage of group B was significantly higher than that in group A (P=0.00); the difference of the percentage between group C and B was not significant (P=0.06), that was also not significant between group C and D (P=0.49). The differences of the adherent rate between K562 and RF/6A (rhesus monkey retinal choroid blood vessel endothelial cell line) among the group A, B, C and D were significant (F=20.62, P=0.00). The adherent rate of group B was significantly higher than that in group A (P=0.00), the adherent rate in group C was significantly lower than that in group B (P=0.01), but it was still significantly higher than that in group A (P=0.002), the difference of adherent rate between group B and D was not significant (P=0.68). Conclusion Under the early stage of diabetic retinopathy, HIF-1α small interfere RNA pSUPERH1-siHIF1α may significantly suppress the expression of CD18 on the surface of K562 cells, but it may not significantly influence the expression of ninjurin-1 on the surface of K562 cells.

Citation： SongHuping, ZhuQi, WuQiong. Hypoxia-inducible factor 1α regulates the expression of CD18 and ninjurin-1. Chinese Journal of Ocular Fundus Diseases, 2014, 30(2): 156-160. doi: 10.3760/cma.j.issn.1005-1015.2014.02.010 Copy

1.	Talahalli R, Zarini S, Tang J, et al. Leukocytes regulate retinal capillary degeneration in the diabetic mouse via generation of leukotrienes[J].J Leukoc Biol, 2013, 93:135-143.
2.	Lutty GA. Effects of diabetes on the eye[J]. Invest Ophthalmol Vis Sci, 2013, 54:81-87.
3.	Serra AM, Waddell J, Manivannan A, et al. CD11b+ bone marrow-derived monocytes are the major leukocyte subset responsible for retinal capillary leukostasis in experimental diabetes in mouse and express high levels of CCR5 in the circulation[J]. Am J Pathol, 2012, 181:719-727.
4.	Li G, Veenstra AA, Talahalli RR, et al. Marrow-derived cells regulate the development of early diabetic retinopathy and tactile allodynia in mice[J]. Diabetes, 2012, 61:3294-3303.
5.	Araki T, Milbrandt J. Ninjurin, a novel adhesion molecule, is induced by nerve injury and promotes axonal growth[J]. Neuron, 1996, 17:353-361.
6.	Ahn BJ, Lee HJ, Shin MW, et al. Ninjurin-1 is expressed in myeloid cells and mediates endothelium adhesion in the brains of EAE rats[J]. Biochem Biophys Res Commun, 2009, 387:321-325.
7.	宋虎平, 惠延年, 雷春灵, 等.缺氧诱导因子1α对早期糖尿病视网膜病变状态下人粒细胞系白血病细胞表面黏附分子CD18表达的调节[J].中华眼底病杂志, 2010, 26:169-172.
8.	宋虎平, 惠延年, 王丽丽.缺氧诱导因子1α对糖尿病大鼠中性粒细胞CD18表达和视网膜白细胞黏附的影响[J].中华眼底病杂志, 2008, 24:268-270.
9.	中华医学会眼科学分会眼底病学组.糖尿病视网膜病变的分期标准[J].眼底病, 1985, 21:113.
10.	许惠卓, 刘双珍, 夏晓波, 等.低氧诱导因子-1α干扰RNA对血管内皮生长因子mRNA表达的抑制作用[J].眼视光学杂志, 2007, 9:228-230.
11.	Berger EA, McClellan SA, Vistisen KS, et al. HIF1α is essential for effective PMN bacterial killing, antimicrobial peptide production and apoptosis in pseudomonas aeruginosa keratitis[J/OL]. PLoS Pathog, 2013, 9:E1003457[2013-07-18]. http://www.plospathogens.org/article/info%3Adoi%2F10.1371%2Fjournal.ppat.1003457.
12.	Yasinska IM, Gibbs BF, Lall GS, et al.The HIF1α transcription complex is essential for translational control of myeloid hematopoietic cell function by maintaining mTOR phosphorylation[J]. Cell Mol Life Sci, 2014, 71:699-710.
13.	Maiguel D, Faridi MH, Wei C, et al. Small molecule-mediated activation of the integrin CD11b/CD18 reduces inflammatory disease[J]. Sci Signal, 2011, 4:57.
14.	Sims TN, Dustin ML. The immunological synapse:integrins take the stage[J]. Immunol Rev, 2002, 186:100-117.
15.	Kebir H, Ifergan I, Alvarez JI, et al. Preferential recruitment of interferon-gamma-expressing TH17 cells in multiple sclerosis[J]. Ann Neurol, 2009, 66:390-402.
16.	Molloy ES, Calabrese LH. Therapy:targeted but not trouble-free:efalizumab and PML[J]. Nat Rev Rheumatol, 2009, 5:418-419.
17.	Lee HJ, Ahn BJ, Shin MW, et al. Ninjurin-1:a potential adhesion molecule and its role in inflammation and tissue remodeling[J]. Mol Cells, 2010, 29:223-227.
18.	宋虎平, 雷春灵, 惠延年, 等.神经损伤诱导蛋白-1对早期糖尿病大鼠视网膜髓样细胞黏附的调节作用[J].中华眼科杂志, 2012, 48:1102-1106.
19.	Ifergan I, Kebir H, Terouz S, et al. Role of ninjurin-1 in the migration of myeloid cells to central nervous system inflammatory lesions[J]. Ann Neurol, 2011, 70:751-763.

1. Talahalli R, Zarini S, Tang J, et al. Leukocytes regulate retinal capillary degeneration in the diabetic mouse via generation of leukotrienes[J].J Leukoc Biol, 2013, 93:135-143.
2. Lutty GA. Effects of diabetes on the eye[J]. Invest Ophthalmol Vis Sci, 2013, 54:81-87.
3. Serra AM, Waddell J, Manivannan A, et al. CD11b+ bone marrow-derived monocytes are the major leukocyte subset responsible for retinal capillary leukostasis in experimental diabetes in mouse and express high levels of CCR5 in the circulation[J]. Am J Pathol, 2012, 181:719-727.
4. Li G, Veenstra AA, Talahalli RR, et al. Marrow-derived cells regulate the development of early diabetic retinopathy and tactile allodynia in mice[J]. Diabetes, 2012, 61:3294-3303.
5. Araki T, Milbrandt J. Ninjurin, a novel adhesion molecule, is induced by nerve injury and promotes axonal growth[J]. Neuron, 1996, 17:353-361.
6. Ahn BJ, Lee HJ, Shin MW, et al. Ninjurin-1 is expressed in myeloid cells and mediates endothelium adhesion in the brains of EAE rats[J]. Biochem Biophys Res Commun, 2009, 387:321-325.
7. 宋虎平, 惠延年, 雷春灵, 等.缺氧诱导因子1α对早期糖尿病视网膜病变状态下人粒细胞系白血病细胞表面黏附分子CD18表达的调节[J].中华眼底病杂志, 2010, 26:169-172.
8. 宋虎平, 惠延年, 王丽丽.缺氧诱导因子1α对糖尿病大鼠中性粒细胞CD18表达和视网膜白细胞黏附的影响[J].中华眼底病杂志, 2008, 24:268-270.
9. 中华医学会眼科学分会眼底病学组.糖尿病视网膜病变的分期标准[J].眼底病, 1985, 21:113.
10. 许惠卓, 刘双珍, 夏晓波, 等.低氧诱导因子-1α干扰RNA对血管内皮生长因子mRNA表达的抑制作用[J].眼视光学杂志, 2007, 9:228-230.
11. Berger EA, McClellan SA, Vistisen KS, et al. HIF1α is essential for effective PMN bacterial killing, antimicrobial peptide production and apoptosis in pseudomonas aeruginosa keratitis[J/OL]. PLoS Pathog, 2013, 9:E1003457[2013-07-18]. http://www.plospathogens.org/article/info%3Adoi%2F10.1371%2Fjournal.ppat.1003457.
12. Yasinska IM, Gibbs BF, Lall GS, et al.The HIF1α transcription complex is essential for translational control of myeloid hematopoietic cell function by maintaining mTOR phosphorylation[J]. Cell Mol Life Sci, 2014, 71:699-710.
13. Maiguel D, Faridi MH, Wei C, et al. Small molecule-mediated activation of the integrin CD11b/CD18 reduces inflammatory disease[J]. Sci Signal, 2011, 4:57.
14. Sims TN, Dustin ML. The immunological synapse:integrins take the stage[J]. Immunol Rev, 2002, 186:100-117.
15. Kebir H, Ifergan I, Alvarez JI, et al. Preferential recruitment of interferon-gamma-expressing TH17 cells in multiple sclerosis[J]. Ann Neurol, 2009, 66:390-402.
16. Molloy ES, Calabrese LH. Therapy:targeted but not trouble-free:efalizumab and PML[J]. Nat Rev Rheumatol, 2009, 5:418-419.
17. Lee HJ, Ahn BJ, Shin MW, et al. Ninjurin-1:a potential adhesion molecule and its role in inflammation and tissue remodeling[J]. Mol Cells, 2010, 29:223-227.
18. 宋虎平, 雷春灵, 惠延年, 等.神经损伤诱导蛋白-1对早期糖尿病大鼠视网膜髓样细胞黏附的调节作用[J].中华眼科杂志, 2012, 48:1102-1106.
19. Ifergan I, Kebir H, Terouz S, et al. Role of ninjurin-1 in the migration of myeloid cells to central nervous system inflammatory lesions[J]. Ann Neurol, 2011, 70:751-763.

Chinese Journal of Ocular Fundus Diseases

Hypoxia-inducible factor 1α regulates the expression of CD18 and ninjurin-1

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