右美托咪定对于尾吊骨质疏松大鼠骨代谢调控作用及机制  

Role and mechanism of dexmedetomidine in regulating bone metabolism in tail-suspended osteoporotic rats

作　　者：郭云亮 王灿[1] 张新雨 颜泽栋 温志鹏 刘若冰 刘鹏森 GUO Yunliang;WANG Can;ZHANG Xinyu;YAN Zedong;WEN Zhipeng;LIU Ruobing;LIU Pengsen(Department of Anesthesiology,Henan Provincial Hospital of Traditional Chinese Medicine,Zhengzhou,Henan;Faculty of Biomedical Engineering,Airforce Medical University,Xi’an,Shaanxi,China)

机构地区：[1]河南省中医院麻醉科,河南郑州 [2]空军军医大学军事生物医学工程学系,陕西西安

出　　处：《陆军军医大学学报》2025年第3期226-233,共8页Journal of Army Medical University

基　　金：国家自然科学基金青年项目(12302412);国家自然科学基金面上项目(52377229)。

摘　　要：目的研究右美托咪定(dexmedetomidine,DEX)对于尾吊大鼠骨丢失的作用效果,并初步探究其对于骨代谢的调控机制。方法30只雄性大鼠按照随机数字表法分为对照(Control)组、尾吊(HU)组、尾吊+右美托咪定(HU+DEX)组,每组10只。HU组和HU+DEX组大鼠行4周后肢悬吊处理,HU+DEX组大鼠从尾吊当天开始进行DEX干预(10μg/kg,隔天1次),Control组和HU组大鼠每日行等量生理盐水注射。通过骨组织学染色检测小梁骨面积占比,通过生物力学三点弯曲测试检测最大载荷、刚度及断裂能,通过钙黄绿素/茜素红荧光双标检测矿化沉积率和骨形成速率,通过抗酒石酸酸性磷酸酶(tartrate-resistant acid phosphatase,TRAP)染色分析骨表面破骨细胞数。分离尾吊大鼠胫骨原代成骨细胞,进行DEX干预(1 nmol/L),通过碱性磷酸酶染色和活性测试检测碱性磷酸酶阳性成骨细胞占比和成骨细胞碱性磷酸酶活性,通过成骨相关基因表达检测成骨活性功能相关因子,包括骨钙素(osteocalcin,Ocn)、Runt相关转录因子2(runt related transcription factor 2,Runx2)、Osterix蛋白(osterix,Osx)和I型胶原蛋白(type 1 collagen,Col1)。结果动物实验结果揭示,DEX显著增加了尾吊大鼠胫骨骨小梁的面积占比(P<0.001),显著抑制尾吊大鼠骨力学强度的降低(P<0.001),显著增加尾吊大鼠小梁骨矿化沉积率和新骨形成速率(P<0.001)。体外细胞实验结果表明,DEX显著增加了尾吊大鼠体外原代成骨细胞碱性磷酸酶活性及碱性磷酸酶阳性成骨细胞数量(P<0.01),并上调了成骨分化相关基因(Ocn、Runx2、Osx、Col1)的表达水平(P<0.01)。结论DEX能对抗尾吊大鼠骨丢失,刺激成骨细胞的骨形成可能是其作用机制。Objective To investigate the effect of dexmedetomidine(Dex)on bone loss in tailsuspended rats and primarily explore its regulatory mechanism on bone metabolism.Methods A total of 30 male rats were randomly divided into a control group,a model group,and a Dex group,with 10 animals in each group.Rat model of osteoporosis was established by hind limb suspension for 4 weeks.Dex at a dose of 10μg/kg was given intraperitoneally,once every other day from the day of tail suspension.And equal amount of normal saline was given to the control and model group.Bone histological staining was used to observe the trabecular bone area fraction.Biomechanical three-point bending test was employed to measure the maximum load,stiffness,and fracture energy.Dual calcein/alizarin red fluorescence labeling and tartrate resistant acid phosphatase(TRAP)staining were applied respectively to detect the mineral apposition rate and bone formation rate as well as the number of osteoclasts on bone surfaces.Secondly,after primary osteoblasts were isolated from the tibiae of tail-suspended rats and then treated with 1 nmol/L Dex,the proportion of alkaline phosphatase(ALP)-positive osteoblasts and the activity of the enzyme were detected by ALP staining and activity test.qRT-PCR was applied to measure the expression of osteogenic activity-related factors,including osteocalcin(Ocn),Runt related transcription factor 2(Runx2),Osterix protein(Osx),and type 1 collagen(Col1).Results The animal experiments revealed that Dex treatment significantly increased the tibial trabecular bone area fraction,inhibited the decrease in bone mechanical strength,and enhanced the mineralization deposition rate and new bone formation rate of trabecular bone in the tail-suspended rats(all P<0.001).The in vitro experiments showed that Dex treatment obviously improved ALP activity and the number of ALP-positive osteoblasts in primary osteoblasts isolated from tail-suspended rats(P<0.01),and up-regulated the expression levels of osteogenic differentiation-related genes,such

关 键 词：右美托咪定 成骨 尾吊大鼠 骨质疏松 成骨细胞 

分 类 号：R322.71[医药卫生—人体解剖和组织胚胎学] R681.02[医药卫生—基础医学] R971.3