细胞膜微粒CD31、CD54与酒精性股骨头缺血性坏死的关系:随机对照实验方案

Relationship of cell membrane microparticles CD31 and CD54 with alcohol-induced avascular necrosis of the femoral head: study protocol for a randomized controlled trial

背景:细胞膜微粒CD31、CD54可通过介导血管炎症反应、促凝、影响血管舒缩及促进血管内皮损伤导致股骨头内微血管损伤。文献已证实细胞膜微粒在激素性股骨头坏死中发挥重要作用,但是目前尚未见微粒与酒精性股骨头缺血性坏死相关性的研究。方法/设计:随机对照动物实验。健康雄性Wistar大鼠随机分为2组,模型组连续6个月灌胃给予烈性白酒制备酒精性股骨头缺血性坏死模型,空白对照组给予等量生理盐水灌胃。干预1-6个月,每月2组各随机取6只大鼠,分批取血,流式细胞仪检测血清细胞膜微粒CD31、CD54水平;取双侧股骨头,固定、脱钙、石蜡包埋切片,行苏木精-伊红染色光镜下进行空骨陷窝计数确定股骨头坏死情况,再行Verhoeff氏染色及MSB微血栓染色观察股骨头内微血管损伤及微血栓形成情况;并分析细胞膜微粒CD31、CD54水平与股骨头坏死、血管内皮损伤及微血栓形成的相关性。讨论:实验结果拟探讨细胞膜微粒CD31、CD54在酒精性股骨头坏死中的作用,更加深入探讨酒精性股骨头坏死发病机制,对其早期诊断、早期治疗提供新的理论依据,可能为其治疗提供一种新的靶点。伦理批准:实验方案经内蒙古医科大学动物实验伦理委员会批准,批准号为YKD2016154。大鼠的实验操作和取材遵循《关于善待实验动物的指导性意见》的规定,并与美国国立卫生与健康研究院的指南一致。

BACKGROUND：Cel membrane microparticles CD31 and CD54 lead to microvascular injury in the femoral head by mediating vascular inflammatory response, promoting blood clotting, affecting vasomotion and promoting vascular endothelial injury. Studies have verified that membrane particles play an important role in steroid-induced necrosis of the femoral head, but there is no studies concerning relationship between microparticles and alcohol-induced avascular necrosis of femoral head. METHODS/DESIGN：This is a randomized control ed animal study. Healthy male Wistar rats wil be randomly assigned to two groups. In the model group, rats wil be intragastrical y administered hard liquor for 6 consecutive months to prepare models of alcohol-induced avascular necrosis of the femoral head. Blank controls wil be intragastrical y given an equal volume of physiological saline. In 1-6 months of intervention, six rats wil be randomly selected from each group every month. Blood wil be col ected separately. Flow cytometry wil be used to detect serum cel membrane particles CD31, CD54 levels. Bilateral femoral head wil be fixed, decalcified, embedded in wax, and then sections. After hematoxylin-eosin staining, empty bone lacuna wil be quantified under a light microscope to identify femoral head necrosis. Verhoff’s staining and MSB microthrombosis staining wil be used to observe microvascular injury and microvascular thrombosis in the femoral head, and to analyze the correlation of CD31 and CD54 levels with femoral head necrosis, vascular endothelial injury and microvascular thrombosis. DISCUSSION：This study wil investigate the effects of CD31 and CD54 on alcohol-induced avascular necrosis of the femoral head, explore the pathogenesis of alcohol-induced avascular necrosis of the femoral head, provide a new theoretical basis for early diagnosis and early treatment, and may provide a new target for its treatment. ETHICS APPROVAL：The protocol has been approved by the Animal Experimental Ethics Committee of Inner Mongolia Medical University （approval number YKD2016154）. Experimental procedures and materials of rats wil be in accordance with the Guide for the Care and Use of Laboratory Animals, which is consistent with the guide of National Institutes of Health.