模拟失重可能通过OPG/RANKL/RANK系统介导引起大鼠脑动脉钙化  

Simulated Weightlessness May Result in Cerebral Artery Calcification in Rats by Altering OPG/RANKL/RANK System

作　　者：刘焕 茹凝玉 白云刚 周颖 武艳玮 徐辉 马进 苏兴利 Liu Huan;Ru Ningyu;Bai Yungang;Zhou Ying;Wu Yanwei;Xu Hui;Ma Jin;Su Xingli(不详;Department of Basic Medicine,Xi'an Medical University,Xi’an Shaanxi 710021,China)

机构地区：[1]西安医学院基础医学部,陕西西安710021 [2]空军军医大学航空航天医学系,陕西西安710032 [3]宁夏医科大学基础医学院病理学系,宁夏银川750004

出　　处：《航天医学与医学工程》2021年第1期1-6,共6页Space Medicine & Medical Engineering

基　　金：国家自然科学基金(81601637,81800211);陕西省重点研发计划项目(2020SF-156);陕西省自然科学基础研究计划项目(2016JQ8001);陕西省呼吸病预防与诊治工程研究中心开放基金(2017GCKF09);西安医学院博士科研启动基金(2017DOC16)。

摘　　要：目的探讨4周和8周不同尾吊时间模拟失重大鼠脑动脉钙化情况及相关机制。方法75只SD大鼠随机分为对照组、4周尾吊组和8周尾吊组3组,每组25只。建立不同尾吊时间的模拟失重大鼠模型,建模成功后分离各组大鼠脑动脉,钙含量检测、茜素红染色和Von Kossa染色观察脑动脉钙盐沉积情况,检测碱性磷酸酶(ALP)活性观察钙代谢相关酶的变化,蛋白免疫印迹实验检测成骨样细胞表型转化相关蛋白、骨保护素(OPG)、核因子κB受体活化因子(RANK)及其配体RANKL的蛋白表达水平。结果与对照组比较,4周和8周尾吊大鼠脑动脉钙含量增加,钙盐沉积增多(P<0.01),ALP活性增强(P<0.01),成骨样细胞表型转化相关蛋白Runx2,Msx2,骨钙素表达增加,收缩表型蛋白SM22α表达降低(P<0.05,P<0.01)。4周尾吊大鼠脑动脉OPG,RANK和RANKL蛋白表达增加。结论4周和8周模拟失重引起大鼠脑动脉平滑肌细胞由收缩表型向成骨样细胞表型转化,并引起动脉钙化,OPG/RANKL/RANK系统可能参与介导模拟失重引起脑动脉钙化的病理过程。Objective To explore the effects of 4-week and 8-week simulated weightlessness by hindlimb unloading(HU) on cerebral artery calcification in rats and its underlying mechanisms.Methods Seventy-five rats were randomly divided into control group,4-week HU group and 8-week HU group(n=25,each).Hindlimb unloading rat model was used to simulate the effects of weightlessness.After successful modeling,the cerebral arteries were separated.The quantification of calcium content,the alizarin red staining and the Von Kossa staining were used to detect the calcium salt deposition.The alkaline phosphatase(ALP) activity was measured to detect the dysfunction of calcium metabolism related enzyme.Western blot analysis was performed to measure the protein expression levels of osteoblast markers,osteoprotegerin(OPG),receptor activator of nuclear factor κB(RANK) and RANK ligand(RANKL).Results Compared with the control group,the calcium content and the calcium salt deposition were increased;the ALP activity was enhanced;the protein expression of osteoblast markers Runx2,Msx2,osteocalcin increased,whereas the expression of systolic phenotype marker SM22α decreased in the cerebral arteries of both 4-week and 8-week HU rats(P<0.05,P<0.01).Moreover,the protein expression of OPG,RANK and RANKL were increased in the cerebral arteries of 4-week HU rats.Conclusion The 4-week and 8-week simulated microgravity could result in osteogenic transdifferentiation of cerebral artery smooth muscle cells and thus vascular calcification in rats,and the dysregulation of OPG/RANKL/RANK system may account for the above changes.

关 键 词：模拟失重 血管钙化 血管平滑肌细胞 骨保护素 核因子ΚB受体活化因子 

分 类 号：R852.22[医药卫生—航空、航天与航海医学]