门冬酰胺酶干预可促进激素诱导模型小鼠股骨头的缺血与坏死
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摘要
背景:门冬酰胺酶能抑制血管生成及影响局部血运,对血管内皮细胞造成伤害,从而影响股骨头局部血运。目的:对比观察门冬酰胺酶对地塞米松诱导缺血性股骨头坏死的影响。方法:Balb/c雄性小鼠由大连医科大学实验动物中心提供,实验方案经大连大学附属中山医院动物实验伦理委员会批准。Balb/c雄性小鼠60只随机分为4组:①门冬酰胺酶+地塞米松组(n=15):小鼠饮水中放入地塞米松2 mg/L,实验一开始该组小鼠腹腔注射门冬酰胺酶(1 200 U/kg);②门冬酰胺酶组(n=15):门冬酰胺酶用量、用法同门冬酰胺酶+地塞米松组;③地塞米松组(n=15),小鼠饮水中放入地塞米松2 mg/L;④对照组(n=15):饲料和饮水中不放任何药物。8周后麻醉下处死小鼠,观察小鼠体质量、凝血因子指标、股骨头组织苏木精-伊红染色及免疫组织化学染色结果。结果与结论:①门冬酰胺酶+地塞米松组小鼠体质量增长明显弱于其他3组;②门冬酰胺酶+地塞米松组小鼠凝血因子Ⅲ和凝血因子Ⅴ明显高于其他3组(P <0.05或P <0.01);③与其他3组比较,门冬酰胺酶+地塞米松组小鼠股骨头空骨陷窝率更高(P <0.05),骨小梁占有率更小(P <0.05),骨坏死程度更加明显;④免疫组织化学门冬酰胺酶+地塞米松组骨保护素积分吸光度值明显大于其他3组(P <0.05或P <0.01);⑤结果说明,门冬酰胺酶能够显著促进小鼠缺血性股骨头坏死。
BACKGROUND: Asparaginase can inhibit angiogenesis and affect blood supply, and make damage to the vascular endothelial cells, thereby affecting the local blood supply of femoral head.OBJECTIVE: To investigate the effect of asparaginase on the avascular necrosis of femoral head induced by dexamethasone.METHODS: Balb/c male mice were provided by Laboratory Animal Center of Dalian Medical University, and the study was approved by the Experimental Ethics Committee of Affiliated Zhongshan Hospital of Dalian University. Sixty male Balb/c mice were randomly divided into four groups: asparaginase + dexamethasone group(n=15, 2 mg/L dexamethasone in the water, and intraperitoneal injection of 1 200 U/kg asparaginase), asparaginase group(n=15, intraperitoneal injection of 1 200 U/kg asparaginase), dexamethasone group(n=15, 2 mg/L dexamethasone in the water), and control group(n=15, no intervention). All mice were killed after 8 weeks. The body mass, coagulation factors, hematoxylin-eosin staining and immunohistochemical staining of the femoral head were observed.RESULTS AND CONCLUSION:(1) The body mass gain in the asparaginase + dexamethasone group was significantly lower than that in the other three groups.(2) The levels of coagulation factor III and V in the asparaginase + dexamethasone group were significantly higher than those in the other three groups(P < 0.05 or P < 0.01).(3) Compared with the other three groups, the empty bone lacunae rate was increased(P < 0.05), and trabecular fracture rate was decreased(P < 0.05), and the degree of osteonecrosis was more obvious in the asparaginase +dexamethasone group.(4) The absorbance value of osteoprotegerin in the asparaginase + dexamethasone group was significantly lower than that in the other three groups(P < 0.05 or P < 0.01).(5) These results indicate that asparaginase can promote avascular necrosis of femoral head in mice.
BACKGROUND: Asparaginase can inhibit angiogenesis and affect blood supply, and make damage to the vascular endothelial cells, thereby affecting the local blood supply of femoral head.OBJECTIVE: To investigate the effect of asparaginase on the avascular necrosis of femoral head induced by dexamethasone.METHODS: Balb/c male mice were provided by Laboratory Animal Center of Dalian Medical University, and the study was approved by the Experimental Ethics Committee of Affiliated Zhongshan Hospital of Dalian University. Sixty male Balb/c mice were randomly divided into four groups: asparaginase + dexamethasone group(n=15, 2 mg/L dexamethasone in the water, and intraperitoneal injection of 1 200 U/kg asparaginase), asparaginase group(n=15, intraperitoneal injection of 1 200 U/kg asparaginase), dexamethasone group(n=15, 2 mg/L dexamethasone in the water), and control group(n=15, no intervention). All mice were killed after 8 weeks. The body mass, coagulation factors, hematoxylin-eosin staining and immunohistochemical staining of the femoral head were observed.RESULTS AND CONCLUSION:(1) The body mass gain in the asparaginase + dexamethasone group was significantly lower than that in the other three groups.(2) The levels of coagulation factor III and V in the asparaginase + dexamethasone group were significantly higher than those in the other three groups(P < 0.05 or P < 0.01).(3) Compared with the other three groups, the empty bone lacunae rate was increased(P < 0.05), and trabecular fracture rate was decreased(P < 0.05), and the degree of osteonecrosis was more obvious in the asparaginase +dexamethasone group.(4) The absorbance value of osteoprotegerin in the asparaginase + dexamethasone group was significantly lower than that in the other three groups(P < 0.05 or P < 0.01).(5) These results indicate that asparaginase can promote avascular necrosis of femoral head in mice.
引文
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[2]Li J,Fan L,Yu Z,etal.The effect of deferoxamineon angiogenesis and bone repair in steroid-inducedosteonecrosis of rabbit femoral heads.Exp Biol Med.2015;240(2):273-280.
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[11]Zeng X,Zhan K,Zhang L,et al.The impact of high total cholesterol and high low-density lipoprotein on avascular necrosis of the femoral head in low-energy femoral neck fractures.J OrthopSurg Res.2017;12(1):30.
[12]Zheng L,Wang W,Ni J,et al.The association of e NOS gene polymorphism with avascular necrosis of femoral head.PloSone.2014;9(2):e87583.
[13]孟晨阳,刘万林,白锐,等.激素性股骨头缺血性坏死发病机制中的细胞自噬[J].中国组织工程研究,2017,21(8):1280-1287.
[14]刘彬,李刚,许波,等.激素性股骨头坏死成脂分化学说及治疗现状[J].中国组织工程研究,2014,18(29):4730-4735.
[15]Beckmann R,Shaheen H,Kweider N,et al.Enoxaparin prevents steroid-related avascular necrosisof the femoral head.Scientifc World Journal.2014;2014:347813.
[16]Ichiseki T,Kaneuji A,Ueda Y,et al.Osteonecrosis development in a novel rat model characterized bya single application of oxidative stres.Arthritis Rheum.2011;63(7):2138-2141.
[17]Boquete-Castro A,Gómez-Moreno G,Calvo-Guirado JL,et al.Denosumab and osteonecrosisof the jaw.A systematic analysis of events reported inclinical trials.Clin Oral Implants Res.2016;27(3):367-375.
[18]Couturier MA,Huguet F,Chevallier P,et al.Cerebral venous thrombosis inadult patients with acute lymphoblastic leukemia or lymphoblastic lymphoma during induction chemotherapy with l-asparaginase:The GRAALL experience.Am J Hematol.2015;90(11):986-991.
[19]Murakawa M,Okamura T,Shibuya T,et al.Use of asyn-thetic protease inhibitor for the treatment of L-asparaginaseinduced acute pancreatitis complicated by disseminated intravascular coagulation.Ann Hematol.1992;64(5):249-252.
[20]Qiang H,Liu H,Ling M,et al.Early steroid-inducedosteonecrosis of rabbit femoral head and panaxnotoginsengsaponins:mechanism and protective efects.Evid Based Complement Alternat Med.2015(1):1-10.
[21]Rupp RE,Rupp SN.Femoral head avascular necrosis isnot caused by arthroscopic posterolateralfemoroplasty.Orthopedics.2016;39(3):177-180.
[22]Jiang Y,Zhang Y,Chen W,et al.Achyranthesbidentata extract exerts osteoprotective effects on steroid-induced osteonecrosis of the femoral head in rats by regulating RANKL/RANK/OPG signaling.J Transl Med.2014;12:334.
[23]Teitelbaum SL.Bone resorption by osteoclasts.Science.289:1504-1508,2000.
[24]Yamada Y,Ando F,Niino N.Association ofpolymorphisms of the osteoprotegerin gene with bone mineraldensity in Japanese women but not men.Mol Genet Metab.2003;80:344-349.
[25]Gori F,Hofbauer LC,Dunstan CR,et al.The expression of osteoprotegerin and RANK ligandand the support of osteoclast formation by stromal-osteoblastlineage cells is developmentally regulated.Endocrinology.2000;141:4768-4776.
[2]Li J,Fan L,Yu Z,etal.The effect of deferoxamineon angiogenesis and bone repair in steroid-inducedosteonecrosis of rabbit femoral heads.Exp Biol Med.2015;240(2):273-280.
[3]Harlev D,Zaidman I,Sarig G,et al.Prophylactic therapy with enoxaparin in children with acute lymphoblastic leukemia and inherited thrombophilia during L-asparaginasetreatment.Thromb Res.2010;126(2):93-97.
[4]Murakawa M,Okamura T,Shibuya T,et al.Use of asyn-thetic protease inhibitor for the treatment of L-asparaginase induced acute pancreatitis complicated by disseminated intravascular coagulation.Ann Hematol.1992;64(5):249-252.
[5]Yang L,Boyd K,Kaste SC,et al.A mouse model for glucocorticoid-induced osteonecrosis:effect of a steroid holiday.J Orthop Res.2009;27(2):169-175.
[6]Hofbauer LC,Neubauer A,Heufelder AE.Receptor activator of nuclear factor-kappa B ligand and osteoprotegerin:Potential implications for the pathogenesis and treatment of malignant bone diseases.Cancer.2001;92(3):460-470.
[7]Singh PP,van der Kraan AG,Xu Jet al.Membrane-bound receptor activator of NFκB ligand(RANKL)activity displayed by osteoblasts is differentially regulated by osteolytic factors.BiochemBiophys Res Commun.2012;422(1):48-53.
[8]Boyce BF,Xing L.Functions of RANKL/RANK/OPG in bone modeling and remodeling.Arch Biochem Biophys.2008;473(2):139-146.
[9]Theoleyre S,Wittrant Y,Tat SK,et al.The molecular triad OPG/RANK/RANKL:Involvement in the orchestration of pathophysiological bone remodeling.Cytokine Growth Factor Rev.2004;15(6):457-475.
[10]Kawedia JD,Janke L,Funk AJ,et al.Substrain-specific differences in survival and osteonecrosis incidence in a mouse model.Comparative medicine.2012;62(6):466-471.
[11]Zeng X,Zhan K,Zhang L,et al.The impact of high total cholesterol and high low-density lipoprotein on avascular necrosis of the femoral head in low-energy femoral neck fractures.J OrthopSurg Res.2017;12(1):30.
[12]Zheng L,Wang W,Ni J,et al.The association of e NOS gene polymorphism with avascular necrosis of femoral head.PloSone.2014;9(2):e87583.
[13]孟晨阳,刘万林,白锐,等.激素性股骨头缺血性坏死发病机制中的细胞自噬[J].中国组织工程研究,2017,21(8):1280-1287.
[14]刘彬,李刚,许波,等.激素性股骨头坏死成脂分化学说及治疗现状[J].中国组织工程研究,2014,18(29):4730-4735.
[15]Beckmann R,Shaheen H,Kweider N,et al.Enoxaparin prevents steroid-related avascular necrosisof the femoral head.Scientifc World Journal.2014;2014:347813.
[16]Ichiseki T,Kaneuji A,Ueda Y,et al.Osteonecrosis development in a novel rat model characterized bya single application of oxidative stres.Arthritis Rheum.2011;63(7):2138-2141.
[17]Boquete-Castro A,Gómez-Moreno G,Calvo-Guirado JL,et al.Denosumab and osteonecrosisof the jaw.A systematic analysis of events reported inclinical trials.Clin Oral Implants Res.2016;27(3):367-375.
[18]Couturier MA,Huguet F,Chevallier P,et al.Cerebral venous thrombosis inadult patients with acute lymphoblastic leukemia or lymphoblastic lymphoma during induction chemotherapy with l-asparaginase:The GRAALL experience.Am J Hematol.2015;90(11):986-991.
[19]Murakawa M,Okamura T,Shibuya T,et al.Use of asyn-thetic protease inhibitor for the treatment of L-asparaginaseinduced acute pancreatitis complicated by disseminated intravascular coagulation.Ann Hematol.1992;64(5):249-252.
[20]Qiang H,Liu H,Ling M,et al.Early steroid-inducedosteonecrosis of rabbit femoral head and panaxnotoginsengsaponins:mechanism and protective efects.Evid Based Complement Alternat Med.2015(1):1-10.
[21]Rupp RE,Rupp SN.Femoral head avascular necrosis isnot caused by arthroscopic posterolateralfemoroplasty.Orthopedics.2016;39(3):177-180.
[22]Jiang Y,Zhang Y,Chen W,et al.Achyranthesbidentata extract exerts osteoprotective effects on steroid-induced osteonecrosis of the femoral head in rats by regulating RANKL/RANK/OPG signaling.J Transl Med.2014;12:334.
[23]Teitelbaum SL.Bone resorption by osteoclasts.Science.289:1504-1508,2000.
[24]Yamada Y,Ando F,Niino N.Association ofpolymorphisms of the osteoprotegerin gene with bone mineraldensity in Japanese women but not men.Mol Genet Metab.2003;80:344-349.
[25]Gori F,Hofbauer LC,Dunstan CR,et al.The expression of osteoprotegerin and RANK ligandand the support of osteoclast formation by stromal-osteoblastlineage cells is developmentally regulated.Endocrinology.2000;141:4768-4776.