| 刘 超,徐 飞,范 新,杨 浩,王蒲春.肌注神经生长因子及血小板衍生生长因子对胫骨干闭合骨折大鼠早期骨愈合的影响及机制探究[J].现代生物医学进展英文版,2023,(9):1642-1646. |
| 肌注神经生长因子及血小板衍生生长因子对胫骨干闭合骨折大鼠早期骨愈合的影响及机制探究 |
| Effect of Intramuscular Injection of Nerve Growth Factor and Platelet-derived Growth Factor on Early Bone Healing in Rats with Closed Tibial Shaft Fracture and Its Mechanism |
| Received:November 07, 2022 Revised:November 30, 2022 |
| DOI：10.13241/j.cnki.pmb.2023.09.008 |
| 中文关键词: 神经生长因子 血小板衍生生长因子 胫骨干闭合骨折 骨愈合 |
| 英文关键词: Nerve growth factor Platelet derived growth factor Closed fracture of tibial shaft Bone healing |
| 基金项目:2017年度陕西省卫生计生委科技基金项目（2017SF-228） |
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| 中文摘要: |
| 摘要 目的：探讨肌肉注射神经生长因子（NGF）和血小板衍生生长因子（PDGF）对胫骨干闭合骨折大鼠早期骨愈合的效果及潜在机制。方法：采用随机数字表法将80只健康成年雄性SD大鼠分为模型组、NGF组、PDGF组和NGF+PDGF组，各20只。建立胫骨干闭合骨折模型后，给予NGF组大鼠肌注0.8 μg NGF；给予PDGF组大鼠肌注0.8 μg PDGF，给予NGF+PDGF组大鼠肌注0.8 μg NGF和0.8 μg PDGF；给予模型组大鼠肌注等体积生理盐水。分别在治疗第2周（T0）、第4周（T1）、第6周（T2）通过X线检查计算骨痂体积，采用酶联免疫吸附法检测血清中碱性磷酸酶（AKP）水平。颈椎脱臼法处死大鼠后采用苏木精-伊红（HE）染色观察胫骨骨折端病理学改变，采用实时荧光定量PCR法检测骨痂组织骨形态发生蛋白2（BMP2）、血管内皮生长因子（VEGF）和胰岛素样生长因子-1（IGF-1）mRNA相对表达水平。结果：NGF+PDGF组大鼠在T1时骨折断端愈合，骨痂体积大于其他三组；NGF组、PDGF组大鼠在T2时骨折断端愈合，骨痂体积大小均大于模型组（P<0.05）。模型组大鼠T2时骨折断端尚未完全愈合，骨痂体积显著大于其他三组（P<0.05）。NGF+PDGF组大鼠T0~T1时血清AKP水平均显著高于其他三组，NGF组和PDGF组大鼠血清AKP水平显著高于模型组（P<0.05）。T2时4组大鼠血清AKP水平比较无显著差异（P>0.05）。T1时，NGF组、PDGF组和NGF+PDGF组大鼠均可见骨小梁形态更加粗大、致密，呈栅栏状排列，骨小梁间的间隙变小，NGF+PDGF组大鼠骨断裂处被新生骨填满，NGF组、PDGF组骨断裂处仍有少量间隙。T1时NGF+PDGF组大鼠BMP2、VEGF和IGF-1相对表达水平均显著高于其他三组（P<0.05），NGF组和PDGF组大鼠各指标mRNA相对表达水平比较无显著差异（P>0.05），但均显著高于模型组（P<0.05）。T2时各组大鼠骨痂组织中BMP2、VEGF和IGF-1 mRNA相对表达水平比较无显著差异（P>0.05）。结论：NGF和PDGF对胫骨干闭合骨折大鼠早期骨愈合有协同促进作用，可能与促进BMP2、VEGF和IGF-1表达上调有关。 |
| 英文摘要: |
| ABSTRACT Objective: To investigate the effect and potential mechanism of intramuscular injection of nerve growth factor (NGF) and platelet derived growth factor (PDGF) on early bone healing in rats with closed tibial shaft fractures. Methods: Eighty healthy adult male SD rats were randomly divided into model group, NGF group, PDGF group and NGF+PDGF group with 20 rats each. After establishing the model of closed tibial shaft fracture, rats in NGF group were given intramuscular injection of 0.8 μg NGF. Rats in PDGF group received intramuscular injection of 0.8 μg PDGF, the rats in NGF+PDGF group were intramuscularly injected with 0.8 μG NGF and 0.8 μg PDGF; The rats in the control group and model group were given intramuscular injection of physiological saline of the same volume. The callus volume was calculated by X-ray examination at the 2nd week (T0), 4th week (T1) and 6th week (T2) of treatment, and the serum alkaline phosphatase (AKP) level was detected by enzyme-linked immunosorbent assay. After the rats were killed by cervical dislocation method, Hematoxylin eosin (HE) staining was used to observe the pathological changes of tibial fracture end, and real-time fluorescence quantitative PCR was used to detect bone morphogenetic protein 2 (BMP2) Relative expression level of vascular endothelial growth factor (VEGF) and insulin like growth factor-1 (IGF-1) mRNA. Results: In NGF+PDGF group, the fracture ends healed at T1, and the callus volume was larger than the other three groups; The fracture ends of NGF group and PDGF group healed at T2, and the volume of callus was larger than that of model group (P<0.05). The fracture ends of the model group were not fully healed at T2, and the callus volume was larger than that of the other three groups (P<0.05). The serum AKP levels of NGF+PDGF group rats were higher than those of the other three groups at T0-T1, and the serum AKP levels of NGF group and PDGF group rats were higher than those of the model group (P<0.05). There was no significant difference in serum AKP level among the four groups at T2 (P>0.05). At T1, the bone trabeculae of rats in NGF group, PDGF group and NGF+PDGF group were thicker, denser, and arranged in a palisade shape. The space between bone trabeculae became smaller. The bone fracture of rats in NGF+PDGF group was filled with new bone, and there was still a small amount of space in the bone fracture of NGF group and PDGF group. At T1, the relative expression level of NGF+PDGF group rats was higher than that of the other three groups (P<0.05). There was no difference between NGF group and PDGF group in the relative expression level of mRNA of each index (P>0.05), but they were higher than that of the model group (P<0.05). At T2, there was no difference in the relative expression levels of BMP2, VEGF and IGF-1 mRNA in the callus tissues of rats in each group (P>0.05). Conclusion: NGF and PDGF can synergistically promote the early bone healing of rats with closed tibial shaft fracture, which may be related to the up-regulated expression of BMP2, VEGF and IGF-1. |
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