Pls3突变相关新型早发骨质疏松大鼠模型的心血管改变及机制  

Cardiovascular changes and its mechanism in a novel rat model of early-onset osteoporosis induced by Pls3 mutation

作　　者：周冰娜 赵笛辰 胡静[1] 张茜[1] 王鸥[1] 夏维波[1] 邢小平[1] 陈太波[3] 李梅[1] ZHOU Bing-na;ZHAO Di-chen;HU Jing;ZHANG Qian;WANG Ou;XIA Wei-bo;XING Xiao-ping;CHEN Tai-bo;LI Mei(Department of Endocrinology,Key Laboratory of Endocrinology,National Health and Family Planning Commission,Peking Union Medical College Hospital,Chinese Academy of Medical Sciences and Peking Union Medical College,Beijing 100730,China;Department of Metabolism and Endocrinology,the Second Xiangya Hospital of Central South University,National Clinical Research Center for Metabolic Diseases,Changsha,Hunan 410011 China;Department of Cardiology,Peking Union Medical College Hospital,Chinese Academy of Medical Sciences&Peking Union Medical College,Beijing 100730,China)

机构地区：[1]中国医学科学院北京协和医院内分泌科,北京100730 [2]中南大学湘雅二医院代谢内分泌科,国家代谢性疾病临床医学研究中心,长沙410011 [3]中国医学科学院北京协和医院心内科,北京100730

出　　处：《中华骨质疏松和骨矿盐疾病杂志》2024年第2期114-122,共9页Chinese Journal Of Osteoporosis And Bone Mineral Research

基　　金：国家重点研发计划(2021YFC2501700);国家自然科学基金面上项目(82070908,82370894);中国医学科学院医学与健康科技创新工程项目(2021-I2M-1-051);中央高水平医院临床研究专项(2022-PUMCH-B-014)。

摘　　要：目的基于自主构建的丝束蛋白3(Plastin 3,PLS3)大片段基因Pls3敲除的新型骨质疏松大鼠模型,评估心血管表型及其与胶原代谢的关系。方法采用CRISPR/Cas9技术构建Pls3基因10~16外显子大片段敲除的大鼠模型(Pls3^(E10-16del/0)),与野生型大鼠进行心脏表型比较,采用免疫组化染色了解PLS3蛋白在心血管的表达,采用超声心动评估大鼠心血管形态及功能,HE染色了解心脏组织学改变,使用扫描电镜评估心脏胶原蛋白的微结构及排列。结果PLS3在野生型大鼠心血管组织细胞质中广泛表达,在Pls3^(E10-16del/0)大鼠心血管的表达明显减少。6~8周龄Pls3^(E10-16del/0)大鼠的心脏及大血管结构与功能未见明显异常,但与野生型大鼠相比,其存在右心室舒张末内径及近端升主动脉内径缩小,血管内皮细胞减少及排列紊乱。Pls3^(E10-16del/0)大鼠心血管组织中的胶原合成未受影响,但电镜下可见其心脏及升主动脉中的细胶原纤维断裂、排列散乱。结论本研究首次对新型Pls3基因大片段敲除的早发骨质疏松大鼠模型心血管系统进行研究,发现其右心室及升主动脉内径缩小,出现细胶原纤维断裂、排列散乱的异常改变,其对心血管系统的结构与功能的远期影响,有待观察,研究拓展了对Pls3相关骨质疏松症心血管表型的认识。Objective To evaluate the cardiovascular phenotype and its relationship to collagen metabolism in a novel rat model of early-onset osteoporosis(EOOP)induced by Pls3 mutation.Methods Comparison of cardiac phenotype was completed between rats with knockout of the 10-16 exons of Pls3(Pls3^(E10-16del/0))constructed by CRISPR/Cas9 technology and wild type rats.Immunohistochemical staining was used to evaluate the expression of PLS3 in cardiovascular tissues.Echocardiography was used to assess cardiovascular morphology and function.H&E staining was used to evaluate the histological change of myocardial tissue.The collagen microstructure and arrangement were evaluated by scanning electron microscopy.Results In the cytoplasm of the cardiovascular tissues,PLS3 was widely expressed in wild-type rats,and significantly decreased in Pls3^(E10-16del/0) rats.At 6-8 weeks of age,no significant structural and functional abnormalities were observed in the heart and great vessels of Pls3^(E10-16del/0) rats.However,compared with wild-type rats,Pls3^(E10-16del/0) rats showed reductions in right ventricular end-diastolic diameter and proximal ascending aorta diameter.In Pls3^(E10-16del/0) rats,collagen synthesis in cardiovascular tissues was not affected,but electron microscopy showed that fine collagen fibers were broken and disorganized in the heart and ascending aorta.Conclusion We firstly evaluated the cardiovascular phenotype of a new Pls3^(E10-16del/0) rat model with EOOP,which had decreased internal diameter of the right ventricle and ascending aorta,broken and scattered fine collagen fibers.The effects of mutation in Pls3 on the cardiovascular system need to be observed for a long time.This study expands the understanding of the cardiovascular phenotype of rare Pls3-associated early-onset osteoporosis.

关 键 词：早发骨质疏松症 Pls3基因突变 心血管表型 

分 类 号：R681[医药卫生—骨科学] R541[医药卫生—外科学]