骨健颗粒促进骨折早期愈合中对VEGF、NOS及ALP的影响

Investigate Gujiankeli(骨健颗粒)affection of VEGF NOS and ALP to promote early fracture healing

目的:观察骨健颗粒在促进骨折愈合早期血管内皮生长因子(VEGF)、一氧化氮合酶(NOS)及其异构体酶活性和碱性磷酸酶(ALP)的活性,探讨三者之间的相关性,阐明骨健颗粒促进骨折早期愈合的机制。方法:50只兔桡骨骨折模型,随机分为模型组、伤科接骨片对照组及骨健颗粒低、中、高剂量组,分别于术后第1、4、7、10、14、21、28d采取外周静脉血,用酶联免疫吸附法和化学比色法及化学分析法测定VEGF的含量和NOS总量、诱导型一氧化氮合酶(iNOS)及原生型一氧化氮合酶(cNOS)的活性和ALP的活性。结果:(1)骨折愈合早期血清中VEGF浓度(以OD值表示),骨健颗粒0.81g/kg剂量组和空白组在1、4、7、14d、21、28d都有显著性差异(P<0.05或P<0.01);骨健颗粒0.81g/kg剂量组和伤科接骨片组比较,在7、14、28d有显著性差异(P<0.05或P<0.01)。(2)血清NOS活性的变化,骨健颗粒0.81g/kg剂量组和伤科接骨片组及空白组在4d、7d、14d时有非常显著的差异(P<0.01);在21d时骨健颗粒0.81g/kg剂量组和空白组有非常显著的差异(P<0.01),在1d时有显著差异(P<0.05)。(3)血清iNOS活性的变化,骨健颗粒0.81g/kg剂量组剂量组和伤科接骨片在7、21d时有显著的差异(P<0.05或P<0.01);骨健颗粒0.81g/kg剂量组剂量组和空白组比较,在1、4、7、21d是有显著的差异(P<0.05或P<0.01)。(4)血清cNOS活性的变化,骨健颗粒0.81g/kg剂量组剂量组和伤科接骨片及空白组在4d、7d、14d时相比有非常显著的差异(P<0.01);骨健颗粒0.81g/kg剂量组剂量组和空白组在1、21d时相比有非常显著的差异(P<0.05或P<0.01)。(5)血清ALP活性的变化,骨健颗粒0.81g/kg剂量组剂量组和伤科接骨片组及空白组在7、14、21、28d相比均有非常显著的差异(P<0.01);骨健颗粒0.81g/kg剂量组剂量组和空白组在4d时相比有非常显著的差异(P<0.01)。以上五组实验结果中,骨健颗粒有效剂量在0.81～1.62g/kg之间,但以0.81g/kg效果为著。结论:骨折愈合早期血清中VEGF、NOS和ALP在不同时段的变化各不相同;骨健颗粒0.81g/kg剂量组能提高VEGF的表达量和激活NOS酶活性以及提高ALP的活性,从而有效的促进了骨折的早期愈合,这可能是其产生临床疗效的机制之一。

Objective：To study the Gujiankeli in the promotion of bone health of early fracture healing of vascular endothelial growth factor（VEGF） nitric oxide synthase（NOS） alkaline phosphatase（ALP） and its isomer activity,to explore the correlation between VEGF NOS and ALP,to further clarify the Gujiankeli to promote bone health of early fracture healing mechanism.Methods：Build 50 rabbits were randomly divided into Gujiankeli group（hight medium low）,Shangkejiegu group and blank group,respectively administration after the first day of taking peripheral 1,4,7,10,14,21,28 day blood,using enzymelinked immunosorbent assay and chemical colorimetric determination of VEGF content total NOS and inducible nitric oxide synthase（iNOS） and nitric oxide synthase native（cNOS） and ALP activity.Results：（1） Healing of early serum VEGF levels（in OD value indicated） in the 1,4,14,21day,Gujiankeli medium dose group and the blank group there was a significant difference（P0.01）,in the 7d significant difference（P0.05）;and Shangkejiegupian group,in the 14d there was a significant difference（P0.01）;in 7d and 28d there is asignificant difference（P0.05）;（2） Serum NOS activity changes in the 4d,7d,14d,when Gujiankeli medium dosegroup group and Shangkejiegupia group and blank group there was a significant difference（P0.01）;in the 21d,when the Gujiankeli medium dose group and the blank group there was a significant difference（P0.01）,while in 1day there are significant differences（P0.05）;（3） Serum iNOS activity changes in 7day,when Gujiankeli medium dose and Shangkejiegu group have a very significant difference（P0.01）,in the 21day,w hen there are significant differences（P0.05）;and the blank group in 7day,21day is a very significant difference（P0.01）,in the 1d,4d,when there are significant differences（P0.05）;（4） Changes in cNOS activity in serum,in 4d,7day,14dya,and when the Gujiankeli medium dose and Shangkejiegu group and blank group subjects compared with bone pieces have a very significant difference（p 0.01）;in the 21day,when Gujiankeli medium dose group and the blank group compared to the very significant differences（P0.01）,in 1day,when there are significant differences（P0.05）.（5） Serum ALP activity of change,Gujiankeli dose group Shangkejiegupian Blank group in 7,14,21,28 day very significant compared difference（P 0.01）;Gujiankeli dose group and blank group group in 4 day when compared with significant difference（P 0.01）.Gujiankeli low group and high dose group compared with other group no obvious difference.Conclusion：Early fracture healing in serum VEGF NOS and ALP in the different changes in different periods;Gujiankeli can improve VEGF expression activation NOS and ALP activity,This may be the generation mechanism of clinical efficacy.