摘要
目的:观察活骨注射液髋关节腔灌注对兔股骨头坏死模型血管内皮生长因子(vascular endothelial growth factor,VEGF)表达的动态影响。方法:将90只新西兰大白兔随机分为对照组、模型组和实验组,每组30只。对模型组和实验组动物采用液氮冷冻法进行股骨头坏死造模;造模成功后,分别以生理盐水和活骨注射液进行髋关节腔灌注。对照组不进行任何干预。药物干预开始后1、3、6、9、12周时,分别从各组随机选取6只动物处死后取出股骨头,采用免疫组化法和实时定量PCR法检测VEGF表达情况。结果:药物干预后各时点,3组动物股骨头VEGF蛋白表达量比较,组间差异均有统计学意义(490.56±60.62,661.99±64.51,796.09±86.17,F=27.670,P=0.000;501.41±58.85,781.71±87.32,854.71±76.97,F=36.810,P=0.000;489.33±53.19,592.42±117.35,905.51±107.95,F=29.930,P=0.000;498.96±40.37,571.31±111.38,1158.20±147.35,F=65.820,P=0.000;500.47±33.67,508.01±138.76,1528.20±146.09,F=150.770,P=0.000)。对照组各时点股骨头VEGF蛋白表达量均低于实验组(P=0.000,P=0.000,P=0.000,P=0.000,P=0.000);药物干预后1、3周时对照组股骨头VEGF蛋白表达量均低于模型组(P=0.001,P=0.000);药物干预后1、6、9、12周时模型组股骨头VEGF蛋白表达量均低于实验组(P=0.005,P=0.000,P=0.000,P=0.000);其余各时点组间两两比较,差异均无统计学意义。药物干预后各时点,3组动物股骨头VEGF mRNA表达量比较,组间差异均有统计学意义(1.000±0.000,1.313±0.380,1.546±0.204,F=9.650,P=0.001;1.000±0.000,1.412±0.345,1.750±0.236,F=19.361,P=0.000;1.000±0.000,1.235±0.095,1.807±0.211,F=35.216,P=0.000;1.000±0.000,1.234±0.250,2.691±0.289,F=137.743,P=0.000;1.000±0.000,1.113±0.180,3.106±0.287,F=293.245,P=0.000)。对照组各时点股骨头VEGF mRNA表达量均低于实验组(P=0.000,P=0.000,P=0.000,P=0.000,P=0.000);药物干预后1、3、6、9周时对照组股骨头VEGF mRNA表达量均低于模型组(P=0.020,P=0.003,P=0.027,P=0.046);药物干预后3、6、9、12周时模型组股骨头VEGF mRNA表达量均低于实验组(P=0.011,P=0.000,P=0.000,P=0.000);其余各时点组间两两比较,差异均无统计学意义。结论:活骨注射液髋关节腔灌注可持续促进兔股骨头坏死模型坏死股骨头局部VEGF表达,这可能是其治疗股骨头坏死的主要作用机制之一。
Objective：To observe the dynamic effect of intra-articular hip injection of Huogu injection on the expression of vascular en-dothelial growth factor（VEGF）in rabbit models with osteonecrosis of the femoral head（ONFH）.Methods：Ninety New Zealand white rab-bits were randomly divided into control group,model group and experimental group,30 cases in each group.The rabbits in model group and experimental group were administrated with liquid-nitrogen refrigeration to build ONFH models,and then they were intra-articular injected with normal saline and Huogu injection respectively in the hip,while the rabbits in the control group were not be treated.At 1 ,3,6,9 and 1 2weeks after the beginning of drug intervention,6 rabbits were randomly selected from each group respectively and were executed,then their femoral heads were fetched out and were sectioned for detecting the expression of VEGF by immunohistochemical method and real-time quantitative PCR method.Results：There was statistical difference in the expression of VEGF protein in femoral head between the 3 groups at each timepoint after the beginning of drug intervention（490.56 ＋/-60.62,661 .99 ＋/-64.51 ,796.09 ＋/-86.1 7,F =27.670,P =0.000;501 .41 ＋/-58.85,781 .71 ＋/-87.32,854.71 ＋/-76.97,F =36.81 0,P =0.000;489.33 ＋/-53.1 9,592.42 ＋/-1 1 7.35, 905.51 ＋/-1 07.95,F =29.930,P =0.000;498.96 ＋/-40.37,571 .31 ＋/-1 1 1 .38,1 1 58.20 ＋/-1 47.35,F =65.820,P =0.000;500.47 ＋/-33.67,508.01 ＋/-1 38.76,1 528.20 ＋/-1 46.09,F =1 50.770,P =0.000）.The expression of VEGF protein in femoral head was lower in control group compared to experimental group at each timepoint after the beginning of drug intervention（P =0.000,P =0.000,P =0.000,P =0.000,P =0.000）.The expression of VEGF protein in femoral head were lower in control group compared to model group at 1 and 3 weeks after the beginning of drug intervention（P =0.001 ,P =0.000）and were lower in model group compared to experi-mental group at 1 ,6,9 and 1 2 weeks after the beginning of drug intervention（P =0.005,P =0.000,P =0.000,P =0.000）,while there was no statistical difference between the paired groups at other timepoints.There was statistical difference in VEGF mRNA expression in the femoral head between the 3 groups at each timepoint after the beginning of drug intervention （1 .000 ＋/-0.000,1 .31 3 ＋/-0.380, 1 .546 ＋/-0.204,F =9.650,P =0.001 ;1 .000 ＋/-0.000.1 .41 2 ＋/-0.345,1 .750 ＋/-0.236,F =1 9.361 ,P =0.000;1 .000 ＋/-0.000, 1 .235 ＋/-0.095,1 .807 ＋/-0.21 1 ,F =35.21 6,P =0.000;1 .000 ＋/-0.000,1 .234 ＋/-0.250,2.691 ＋/-0.289,F =1 37.743,P =0.000;1 .000 ＋/-0.000,1 .1 1 3 ＋/-0.1 80,3.1 06 ＋/-0.287,F =293.245,P =0.000）.The VEGF mRNA expression in the femoral head was lower in control group compared to experimental group at each timepoint after the beginning of drug intervention（P =0.000,P =0.000,P =0.000,P =0.000,P =0.000）,and were lower in control group compared to model group at 1 ,3,6 and 9 weeks after the begin-ning of drug intervention（P =0.020,P =0.003,P =0.027,P =0.046）,and were lower in model group compared to experimental group at 3,6,9 and 1 2 weeks after the beginning of drug intervention（P =0.01 1 ,P =0.000,P =0.000,P =0.000）,while there was no statistical difference between the paired groups at other timepoints.Conclusion：Intra-articular hip injection of Huogu injection can promote the ex-pression of VEGF in necrotic femoral head of rabbit ONFH model,which may be one of the mechanisms of action for treatment of ONFH.
出处
《中医正骨》
2015年第8期1-6,共6页
The Journal of Traditional Chinese Orthopedics and Traumatology
基金
国家自然科学基金项目(81173276)
哈尔滨市科技创新人才项目(2011RFXYS070)
关键词
股骨头坏死
血管内皮生长因子类
活骨注射液
模型
动物
动物实验
femur head necrosis
vascular endothelial growth factors
Huogu injection
models,animal
animal experimentation
