细胞膜微粒与兔激素性股骨头缺血坏死发生机制及药物干预的实验研究

【作者】 纪春良；

【导师】 翁习生；

【作者基本信息】 中国协和医科大学 ， 临床医学， 2008， 博士

【摘要】 【目的】探讨在激素诱发血液高凝、易栓状态导致的股骨头缺血坏死发病过程中,内皮细胞和血小板源性膜微粒的数量以及炎症指标变化。同时在给予干预药物情况下,观察上述膜微粒的数量变化、凝血及炎症指标的改善情况。【方法】健康成年新西兰兔27只,随机分为3组:A组为对照组(n=9),给予与实验组相同剂量的生理盐水肌注1次;B组为实验组(n=10),给予甲泼尼龙20mg/kg肌注射1次;C组为干预组(n=8),连续给予干预药物2.13 g/kg灌胃12天,于灌胃第7天时给予甲泼尼龙20mg/kg肌注射1次。于注射前、注射后1天、3天、7天、14天和28天各抽静脉全血。使用流式细胞仪对CD31+/CD42b-和CD31+/CD42b+的膜微粒进行定量测定。使用Array-ELISA方法测定TNF-α和IL-1β。同时测定抗凝血酶-Ⅲ(AT-Ⅲ)、蛋白C(PC)、纤维蛋白原(FIB)。于第4周时,处死B组3只动物,取双侧股骨头组织学检查,HE染色。【结果】给予激素肌注后,CD31+/cD42b-和CD31+/CD42b+的膜微粒显著升高(p＜0.05),分别在7天和14天达峰。而干预组的膜微粒在激素和干预药物的共同作用下虽有波动,但未出现显著差异。给予激素后,B组的PC、ATIII相对于基线出现显著下降,但与对照组比较无显著差异,FIB较基线升高,注射后7天达峰,且高于对照组。而在干预组中,各时间点相对于基线和对照组无显著变化。B组和C组的TNF-α和IL-1β相对于基线表现出不同程度的升高,且均高于对照组。【结论】应用大剂量糖皮质激素后,兔体内内皮细胞和血小板源性膜微粒数量显著增加,具前凝血潜能的微粒数量的增加可能是大剂量糖皮质激素诱发血液高凝易栓、导致股骨头缺血坏死发生的原因之一。膜微粒直接或间接所致的炎症反应也在股骨头坏死的发生中起到重要作用。干预药物能够抑制微粒的产生,或许有助于预防或避免激素诱发股骨头缺血坏死的发生。更多还原

【Abstract】 Objective:To investigate the alterations of endothelial- and platelet-derived microparticles and the change of coagulation and inflammation in the steroid-induced avascular osteonecrosis of femoral head with/without an intervention agent.Method:Twenty-seven healthy New Zealand rabbits were included and randomized into 3 group:Group A(the control group,n=9) would be injected once intramuscularly with the same dose of 0.9%NaCl solution as the steroid group;Group B(the steroid group,n=10) would be injected once intramuscularly with 20 mg/kg of methylprednisolone;Group C(the intervention group,n=8) would receive a intervention agent continuously for 12 days and a injection of 20 mg/kg methylprednisolone once intramuscularly on the day 7 of intervention.The blood sample should be collected before injection of methylprednisolone and day 1,day 3, day 7,day 14 and day 28 after the injection.The microparticles expressing CD31+/CD42b-and CD31+/CD42b+ would be measured using flow cytometry.And the antithrombin-Ⅲ(AT-Ⅲ),protein C(PC) and fibrinogen(FIB) were also examined. Tumor necrosis factor alpfa(TNF-α) and interleukin 1 beta(IL-1β) would be measured using Array-ELISA.Three rabbits from the steroid group were sacrificed and their femoral heads were taken out to undergo histopathologic examination with HE staining in 4 weeks after the injection.Results:The AT-Ⅲ,PC levels decreased significantly 7 to 28 days and the FIB level increased on day 7 without significance and then decreased slightly after the injection. The numbers of EMP and PMP were also increased markedly after steroid administration.Both TNF-αand IL-1βlevels increased relative to the baseline, and were greater than the control group at each time point,but no significance was observed.No positive finding was observed in the histopathologic examination.Conclusion:High-dose glucocorticosteroid increases the levels of PMP and EMP that may be related to hypercoagulability,thrombosis and inflammation in microcirculation and play an important role in steroid-induced osteonecrosis.The intervention agent appeared to have the efficacy to decrease the MP levels and improve the hypercoagulability and thrombosis.MP can be considered a true target in the pharmacological control of steroid-induced osteonecrosis.更多还原