Hypoxia-inducible factor 1 alpha and granulocyte-macrophage colony stimulating factor in knee osteoarthritis cartilage

doi:10.3969/j.issn.2095-4344.2015.11.009

Abstract

Abstract:

BACKGROUND: Hypoxia-inducible factor 1α and granulocyte-macrophage colony stimulating factor can promote the development of osteoarthritis under inflammatory conditions.

OBJECTIVE: To explore the expression of hypoxia inducible factor 1α and granulocyte-macrophage colony-stimulating factor in knee osteoarthritis cartilage tissue and to analyze its clinical significance.

METHODS: Thirty-two patients with knee osteoarthritis who underwent knee replacement were selected and cartilage tissues were obtained as knee osteoarthritis group. Another 10 patients with femoral neck fractures served as controls. mRNA and protein levels of hypoxia inducible factor 1α and granulocyte-macrophage colony-stimulating factor in the cartilage tissue were detected by real-time PCR and western blot, respectively. Modified Mankin score was used to assess knee damage in knee osteoarthritis patients. Relationship between mRNA levels of hypoxia inducible factor 1α and granulocyte-macrophage colony-stimulating factor and modified Mankin scores were analyzed.

RESULTS AND CONCLUSION: Hypoxia inducible factor 1α and granulocyte-macrophage colony-stimulating factor gene and protein expressions were significantly higher in the knee osteoarthritis group than the control group, in knee osteoarthritis patients aged > 60 years than patients ≤ 60 years (P < 0.05), as well as higher in secondary knee osteoarthritis patients than primary osteoarthritis patients (P < 0.05). Hypoxia inducible factor 1α and granulocyte-macrophage colony-stimulating factor protein levels and gene levels had statistically significant differences between different pathological grades (P < 0.05); moreover, the severer osteoarthritis, the higher expression levels. Hypoxia inducible factor 1α and granulocyte-macrophage colony-stimulating factor mRNA expression of knee osteoarthritis patients were positively correlated with the modified Mankin scores (r=0.83, P < 0.05; r=0.80, P < 0.05). These findings indicate that hypoxia inducible factor 1α and granulocyte-macrophage colony-stimulating factor expression levels are increased in the knee cartilage tissue of osteoarthritis patients, which display a distribution difference among a variety of pathological parameters and play an important role in the pathogenesis of knee osteoarthritis.

中国组织工程研究杂志出版内容重点：组织构建；骨细胞；软骨细胞；细胞培养；成纤维细胞；血管内皮细胞；骨质疏松；组织工程

全文链接：

Key words: Knee, Osteoarthritis, Granulocyte-Macrophage Colony-Stimulating Factorn

CLC Number:

R318

Yu Yang, Cui Lei. Hypoxia-inducible factor 1 alpha and granulocyte-macrophage colony stimulating factor in knee osteoarthritis cartilage [J]. Chinese Journal of Tissue Engineering Research, 2015, 19(11): 1683-1687.

Figures/Tables 2

2.1 两组患者基线资料分析见表1。 2.2 两组软骨标本中低氧诱导因子1α和粒细胞-巨噬细胞集落刺激因子的表达水平比较两组患者低氧诱导因子1α和粒细胞-巨噬细胞集落刺激因子的基因及蛋白表达水平见表2和图1，可见骨性关节炎组患者低氧诱导因子1α和粒细胞-巨噬细胞集落刺激因子的基因及蛋白表达水平均显著高于对照组(P < 0.05)。 2.3 膝关节骨性关节炎不同病理参数与低氧诱导因子1α和粒细胞-巨噬细胞集落刺激因子的表达不同性别、侧别的骨性关节炎患者低氧诱导因子1α和粒细胞-巨噬细胞集落刺激因子的mRNA和蛋白水平差异均无显著性意义(P > 0.05)。而年龄> 60岁的骨性关节炎患者其低氧诱导因子1α的mRNA和蛋白水平，粒细胞-巨噬细胞集落刺激因子的基因水平显著高于≤ 60岁的骨性关节炎患者(P < 0.05)。继发性的骨性关节炎患者2种因子的基因和蛋白水平均显著高于原发性的骨性关节炎患者(P < 0.05)。病程> 5年的患者2种因子基因及蛋白水平均显著高于病程≤5年的患者(P < 0.05)。不同改良Mankin病理评分分级之间，低氧诱导因子1α和粒细胞-巨噬细胞集落刺激因子的基因和蛋白水平差异均有显著性意义(P < 0.05)，程度越重，其表达水平越高。见表3。 2.4 骨性关节炎组低氧诱导因子1α和粒细胞-巨噬细胞集落刺激因子的mRNA表达与改良Mankin评分相关性经Pearson相关分析，骨性关节炎组患者低氧诱导因子1α和粒细胞-巨噬细胞集落刺激因子的mRNA相对表达量与改良Mankin评分之间均存在正相关(r=0.83，P < 0.05；r=0.80，P < 0.05)。以改良Mankin评分为因变量，低氧诱导因子1α mRNA相对表达量为自变量，行直线回归分析，结果显示两者的直线回归方程为：改良Mankin评分=0.09×低氧诱导因子1α+1.29，方程具有统计学意义(F=64.01，P=0.000，r2=0.681，r=0.83)，具有良好的依存关系。以改良Mankin评分为因变量，粒细胞-巨噬细胞集落刺激因子的mRNA相对表达量为自变量，行直线回归分析，结果显示两者的直线回归方程为：改良Mankin评分=0.51×粒细胞-巨噬细胞集落刺激因子+1.12，方程具有显著性意义(F=52.25，P=0.000，r2=0.635，r=0.80)，具有良好的依存关系。 2.5 骨性关节炎组患者低氧诱导因子1α和粒细胞-巨噬细胞集落刺激因子表达的相关性经Pearson相关分析，骨性关节炎组患者低氧诱导因子1α和粒细胞-巨噬细胞集落刺激因子的mRNA相对表达量之间均存在正相关(r=0.79，P < 0.05)，其蛋白表达之间也存在正相关(r=0.72，P < 0.05)。"

References

[1] Wu L, Huang X, Li L,et al.Insights on biology and pathology of hif-1alpha/-2alpha, tgfbeta/bmp, wnt/beta-catenin, and nf-kappab pathways in osteoarthritis. Curr Pharm Des. 2012; 18(22):3293-3312.
[2] 李笑颜,赵建,曾文,等.晚期骨关节炎患者关节软骨细胞中HIF-1α和VEGF的表达[J].上海交通大学学报:医学版,2010,30(7): 812-816.
[3] Pfander D, Swoboda B, Cramer T. The role of hif-1alpha in maintaining cartilage homeostasis and during the pathogenesis of osteoarthritis. Arthritis research & therapy.2006;8:104.
[4] van der Sluijs JA, Geesink RG, van der Linden AJ，et al. The reliability of the Mankin score for osteoarthritis. J Orthop Res. 1992;10(1):58-61.
[5] Pfander D, Gelse K.Hypoxia and osteoarthritis: How chondrocytes survive hypoxic environments. Curr Opin Rheumatol. 2007;19(5):457-462.
[6] Murab S, Chameettachal S, Bhattacharjee M,et al. Matrix-embedded cytokines to simulate osteoarthritis-like cartilage microenvironments. Tissue engineering Part A. 2013;19(15-16):1733-1753.
[7] Hoff P, Buttgereit F, Burmester GR,et al.Osteoarthritis synovial fluid activates pro-inflammatory cytokines in primary human chondrocytes. Int Orthop. 2013;37(1):145-151.
[8] Zhang SL, Liu HQ, Xu XZ,et al. Effects of exercise therapy on knee joint function and synovial fluid cytokine levels in patients with knee osteoarthritis. Molecular medicine reports. 2013; 7(1):183-186.
[9] Cook AD, Pobjoy J, Steidl S, et al. Granulocyte-macrophage colony-stimulating factor is a key mediator in experimental osteoarthritis pain and disease development. Arthritis Res Therapy.2012;14:R199.
[10] Loeser RF. Osteoarthritis year in review 2013: biology. Osteoarthritis Cartilage. 2013;21(10):1436-1442.
[11] Wenham CY, Conaghan PG. New horizons in osteoarthritis. Age and ageing. 2013;42(3):272-278.
[12] 彭勇,陈勇,邬秀娣,等. HIF-1α及VEGF 在类风湿关节炎成纤维样滑膜细胞中的表达及意义[J]. 浙江医学,2013,35(18): 1628-1631.
[13] Ryu JH, Chae CS, Kwak JS,et al. Hypoxia-inducible factor-2alpha is an essential catabolic regulator of inflammatory rheumatoid arthritis. PLoS biology. 2014;12(6):e1001881.
[14] Mendes SO, dos Santos M, Peterle GT,et al. HIF-1alpha expression profile in intratumoral and peritumoral inflammatory cells as a prognostic marker for squamous cell carcinoma of the oral cavity. PLoS One.2014;9(1):e84923.
[15] Rathnasamy G, Ling EA, Kaur C. Hypoxia inducible factor-1alpha mediates iron uptake which induces inflammatory response in amoeboid microglial cells in developing periventricular white matter through MAP kinase pathway. Neuropharmacology.2014;77:428-440.
[16] Hirohata S, Nagai T, Asako K, et al. Induction of type b synoviocyte-like cells from plasmacytoid dendritic cells of the bone marrow in rheumatoid arthritis and osteoarthritis. Clin Immunol.2011;140:276-283.
[17] Atawia RT, Mosli HH, Tadros MG,et al. Modulatory effect of silymarin on inflammatory mediators in experimentally induced benign prostatic hyperplasia: emphasis on PTEN, HIF-1alpha, and NF-kappaB. Naunyn Schmiedebergs Arch Pharmacol. 2014; 387(12):1131-1140.
[18] 吴振彪,卢宁,王彦宏,等.GM-CSF对类风湿性关节炎患者单核细胞合成MMP的影响[J].现代免疫学，2005, 25(5):394-397.
[19] 张延辉,高春阳,李少华.骨性关节炎患者退变软骨及滑膜组织中细胞因子的表达[J].中国组织工程研究,2013,17(37): 6671-6675.
[20] Yudoh K, Nakamura H, Masuko-Hongo K, et al.Catabolic stress induces expression of hypoxia-inducible factor (hif)-1 alpha in articular chondrocytes: Involvement of hif-1 alpha in the pathogenesis of osteoarthritis. Arthritis research & therapy. 2005;7:R904-914.
[21] Pfander D, Cramer T, Swoboda B. Hypoxia and hif-1alpha in osteoarthritis. Int Orthop.2005;29:6-9.
[22] Schipani E, Ryan HE, Didrickson S，et al. Hypoxia in cartilage: HIF-1alpha is essential for chondrocyte growth arrest and survival. Genes & development.2001; 15(21):2865-2876.
[23] Nakao K, Kubota S, Doi H, et al. Collaborative action of m-csf and ctgf/ccn2 in articular chondrocytes: Possible regenerative roles in articular cartilage metabolism. Bone.2005;36:884-892.
[24] Yoshimatsu T, Saitoh A, Ryu JN,et al.Characterization of immortalized human chondrocytes originated from osteoarthritis cartilage. Int J Mol Med. 2001;8(4):345-351.
[25] 徐苏洋,陈永强.低氧诱导因子与骨关节炎[J].国际骨科学杂志,2011,32(6):384-385.