Expression of hypoxia-inducible factor 1 alpha and core binding factor alpha 1 in rat models of femoral fracture combined with cerebral trauma

doi:10.3969/j.issn.2095-4344.2015.18.003

Abstract

Abstract:

BACKGROUND: The low oxygen environment after femoral fracture and cerebral trauma will induce series of related cytokines expression, including hypoxia-inducible factor 1α and core binding factor α1, which play key roles in regulating bone healing. However, whether the accelerated bone healing is correlated with the expression of hypoxia-inducible factor 1α and core binding factor α1 is still unknown.
OBJECTIVE: To construct rat models of brain injury, to compare the expression level of hypoxia-inducible factor 1α and core binding factor α1 in femoral fracture combined with cerebral trauma rats and simple femoral fracture rats, and to assess the influence of cerebral trauma on bone healing.
METHODS: Rats were randomly divided into blank group, simple femoral fracture group and femoral fracture combined with cerebral trauma group. At 1, 2, 3 and 5 weeks after modeling, rats were executed. Bone healing was evaluated using femoral fracture end X-ray score and hematoxylin and eosin staining at callus tissues. Besides, the expression levels of hypoxia-inducible factor 1α and core binding factor α1 of three groups were
determined with immunohistochemistry.
RESULTS AND CONCLUSION: Bone healing in the femoral fracture combined with cerebral trauma group was better than that of simple femoral fracture group. There was significant difference in the expression level of hypoxia-inducible factor 1α and core binding factor α1 between the simple femoral fracture group and femoral fracture combined with cerebral trauma group (P < 0.05). At the same time, the level of simple femoral fracture group and femoral fracture combined with cerebral trauma group was significantly higher than that of blank group, and that in femoral fracture combined with cerebral trauma group was significantly higher than that of simple femoral fracture group (P < 0.05). Results verified that the expression levels of hypoxia-inducible factor 1α and core binding factor α1 of rats with femoral fracture combined with cerebral trauma were significantly high, which may be the major reason why the bone healing was accelerated after fracture combined with brain injury.

中国组织工程研究杂志出版内容重点：肾移植；肝移植；移植；心脏移植；组织移植；皮肤移植；皮瓣移植；血管移植；器官移植；组织工程

全文链接：

Key words: Tissue Engineering, Femur, Fractures, Bone, Fracture Healing

CLC Number:

R318

Bo Xiao-jin, Xu Lin, Luo Xu-dong, Liu Fu-ying, Huang Wen-liang, Guo Yuan, Ma Li-kun, Cheng Xiao-ju, Bo Meng. Expression of hypoxia-inducible factor 1 alpha and core binding factor alpha 1 in rat models of femoral fracture combined with cerebral trauma[J]. Chinese Journal of Tissue Engineering Research, 2015, 19(18): 2800-2806.

Figures/Tables 3

2.4 股骨骨折合并脑损伤模型大鼠骨痂指标变化各组大鼠骨痂苏木精-伊红染色结果见图4。结果显示，单纯股骨骨折组骨折愈合过程明显，造模后1周可见成纤维细胞长入，2周可见骨折断端有肉芽组织继续生长和纤维性骨痂、软骨性骨痂出现，3周可见少量骨小梁，5周可见较成熟骨小梁。股骨骨折合并脑损伤组骨骼愈合过程更加明显，造模后1周看可见肉芽组织生长连接，2周有软骨岛形成，出现大量软骨细胞，3周出现纤维骨痂和软骨骨痂，形成少量编织骨；造模后5周出现大量成熟骨小梁，编织骨继续增加，散落少量肥大软骨细胞。各组免疫组织化学检测低氧诱导因子1α和核心结合因子α1表达情况见图4。低氧诱导因子1α 各组各个时间段均有表达，单纯股骨骨折组低氧诱导因子1α表达呈递增趋势，高峰出现在造模后第2周，第5周时仍高于空白组；股骨骨折合并脑损伤组造模后一开始上升幅度较单纯股骨骨折组明显，2周达高峰，3周后逐渐下降，5周后仍高于空白组；空白组各时间段变化不大，表达不明显，差异无显著性意义(P > 0.05)。同一时间各组表达情况相比，单纯股骨骨折组、股骨骨折合并脑损伤组表达均明显高于空白组，股骨骨折合并脑损伤组明显高于单纯股骨骨折组(P < 0.05)；单纯股骨骨折组、股骨骨折合并脑损伤组内不同时间段数据间比较，差异有显著性意义(P < 0.05)，见表2；证明大鼠分别在骨折和骨折合并脑损伤后早期低氧诱导因子1α就有明显的表达，并有一定的规律。核心结合因子α1各组各个时间段均有表达，单纯股骨骨折组核心结合因子α1表达呈递增趋势，高峰出现在造模后第2，5周时仍高于空白组；股骨骨折合并脑损伤组造模后开始上升幅度更大，2周达高峰，3周后逐渐下降，仍保持较高的表达，5周后仍明显高于空白组；空白组在各时间段表达不明显(P > 0.05)。同一时间各组表达情况相比，单纯股骨骨折组、股骨骨折合并脑损伤组表达均明显高于空白组，股骨骨折合并脑损伤组明显高于单纯股骨骨折组 (P < 0.05)；单纯股骨骨折组、股骨骨折合并脑损伤组不同时间段数据间比较差异有显著性意义(P < 0.05)，见表3。大鼠分别在骨折和骨折合并脑损伤后早期核心结合因子α1就有明显的表达，并有一定的规律。骨折合并脑损伤时核心结合因子α1的表达更为明显，且表达的时间持续较长。"

References

[1] 罗涛,邓廉夫.低氧诱导因子-1在骨折愈合过程中的调控作用[J].国外医学骨科学分册,2005,26(4):240-242.
[2] 刘志奎,张柳,姜晓华,等.脑损伤对骨折愈合中血管内皮生长因子表达的影响[J].中国康复理论与实践,2009,15(3):233-235.
[3] 李建华,李建民.HIF-1α 在脑损伤中的作用及其机制的研究进展[J].滨州医学院学报,2009,32(5):367-369, 373.
[4] 刘晓东,邓廉夫,王君,等.低氧诱导因子1α对成骨细胞功能的调控作用[J].上海交通大学学报:医学版,2007,27(3):298-302.
[5] Li C, Mori S, Li J, et al. Long-term effect of incadronate disodium (YM-175)on fracture healing of femoral shaft in growing rats. J Bone Miner Res. 2001;16(3):429-436.
[6] Marmarou A, Montasser A, Foda A, et al. A new model of diffuse brain injury in rats. J Neuronal. 1994;296:654-673.
[7] Goldberg VM, Powell A, Shaffer JW, et al. Bone grafting: role of histocompatibility in transplantation. Orthop Res. 1985;3(4): 389-404.
[8] Mavrofrydi O, Papazafiri P. Hypoxia-inducible factor-lα increase is an early and sensitive marker of lung cells responding to benzo[a]pyrene. J Environ Pathol Toxicol Oncol. 2012;31(4):335-347.
[9] Chen YR, Dai AG, Hu RC, et al. The expression of hypoxia-inducible factor-1alpha and its hydroxylases in pulmonary arteries of patient with chronic obstructive pulmonary disease. Zhongguo Ying Yong Sheng Li Xue Za Zhi. 2012;28(3):234-238.
[10] Kelley MR, Parsons SH. Redox regulation of the DNA repair function of the human AP endonuclease Ape1/ref-1. Antioxid Redox Signal. 2001;3(4):671-683.
[11] 刘振东,高敏伟,钟杰林,等.2型糖尿病大鼠骨髓腔内过氧化物酶体增殖物激活受体γ和核心结合因子α1 mRNA表达与骨折的愈合[J].中国组织工程研究与临床,2009,13(15):2891-2895.
[12] Dai Z, Guo F, Wu F, et al. Integrin αvβ3 mediates the synergetic regulation of core-binding factor α1 transcriptional activity by gravity and insulin-like growth factor-1 through phosphoinositide 3-kinase signaling. Bone. 2014;69(12):126-132.
[13] Hong YY, Yu FY, Qu JF, et al. Fibroblasts regulate variable aggressiveness of syndromic keratocystic and non-syndromic odontogenic tumors. J Dent Res. 2014;93(9):904-910.
[14] Duan X, Xu H, Wang Y, et al. Expression of core-binding factor α1 and osteocalcin in fluoride-treated fibroblasts and osteoblasts. J Trace Elem Med Biol. 2014;28(3):278-283.
[15] Houser J, Komarek J, Kostlanova N, et al. A soluble fucose-specific lectin from Aspergillus fumigatus conidia-- structure, specificity and possible role in fungal pathogenicity. PLoS One. 2013;8(12):e83077.
[16] Pei Z,Zhang F, Niu Z, et al. Effect of icariin on cell proliferation and the expression of bone resorption/formation-related markers in human periodontal ligament cells. Mol Med Rep. 2013;8(5):1499-1504.
[17] Takarada T, Hinoi E, Nakazato R, et al. An analysis of skeletal development in osteoblast-specific and chondrocyte-specific runt-related transcriptionfactor-2 (Runx2) knockout mice. J Bone Miner Res. 2013;28(10):2064-2069.
[18] Boes M, Kain M, Kakar S, et al. Osteogenic effects of traumatic braininjury on experiment fracture-healing. J Bone Joint Surg. 2006;88(4):738-743.
[19] Ostrowski RP, Colohan AR, Zhang JH. Mechanisms of hyperbaric oxygen induced neuroprotect ion in a rat model of subarachnoid hemorrhage. J Cereb Blood Flow Metab. 2005; 25(5):554.
[20] Gerber HP, Condoreeli F, Park J, et al. Differential transcriptional regulation of the two vascular endothelial growthfactor receptor genes. Fit-J.butnot T FIk-1/KDR, is upregulated by hypoxia. J Biol Chem. 1997;272(38):23659-23667.
[21] Bergeron M, Gidday JM, Yu AY, et al. Role of hypoxia inducible factor in hypoxia-induced ischemic tolerance in neonatal rat brain. Ann Neurol. 2000;48(3):285.
[22] Gaur T, Lengner CJ, Hovhannisyan H, et al. Canonical WNT signaling promotes osteogenesis by directlyst imulating Runx2 gene expression. J Biolchem. 2005;280(39): 33132-33140.
[23] Jones NM, Lee EM, Brown TCS, et al. Hypoxic preconditioning produces differential expression of hypoxia-inducible factor-lalpha (HTF-lalpha) and its regulatory enzyme HTF poly 1 hydroxylase 2 in neonatal rat brain. Neurosci Lett. 2006;404 (1-2): 72-77.
[24] 邓廉夫,冯伟,张玥,等.诱导成纤维细胞表达核结合因子的研究[J].中华外科杂志,2002,40(8):35-38.
[25] 张劲娥,王淑红,张忻,等.低氧诱导因子1α慢病毒载体转染骨髓间充质干细胞的成骨基因表达[J].中国组织工程研究,2014, 18(1):57-62.
[26] 戚金威,程景林,周姝,等.低氧诱导因子1α高表达对急性心肌梗死大鼠骨髓间充质干细胞的动员[J].中国组织工程研究,2014, 18(10):1579-1584.
[27] 惠春影,肖洪艳,何欣玲,等.低氧诱导因子1α基因三点突变体体外转染骨髓间充质干细胞[J].中国组织工程研究,2014,18(19): 2953-2960.
[28] Chen Y, Zhu HB, Liao J, et al. Regulation of naotai recipe on the expression of HIF-lα/VEGF signaling pathway in cerebral ischemia/reperfusion rats. Zhongguo Zhong Xi Yi. 2014; 34(10):1225-1230.
[29] Huang H, Yu J, Yu D, et al. Effects of chemically induced hypoxia on in vitro expression of hypoxia inducible factor-lα, vascular endothelial growth factor, aggrecanase-1, and tissue inhibitor of metalloproteinase-3 in rat mandibular condylar chondrocytes. J Oral Facial Pain Headache. 2014;28(3):269-276.
[30] Chai DM, Bao ZQ, Hu JG, et al. Vasculogenic mimicry and aberrant expression of HIF-lα/E-cad are associated with worse prognosis of esophageal squamous cell carcinoma. J Huazhong Univ Sci Technolog Med Sci. 2013;33(3):385-391.