【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（Pls3E10-16del/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Pls3E10-16del/0大鼠心血管的表达明显减少。6～8周龄Pls3E10-16del/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。Pls3E10-16del/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。更多还原

【Abstract】 Objective To evaluate the cardiovascular phenotype and its relationship to collagen metabolism in a novel rat model of early-onset osteoporosis(EOOP) induced byPls3mutation.Methods Comparison of cardiac phenotype was completed between rats with knockout of the 10-16 exons ofPls3(Pls3E10-16del/0) constructed by CRISPR/Cas9technology 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 inPls3E10-16del/0rats. At 6-8 weeks of age, no significant structural and functional abnormalities were observed in the heart and great vessels ofPls3E10-16del/0rats. However, compared with wild-type rats,Pls3E10-16del/0rats showed reductions in right ventricular end-diastolic diameter and proximal ascending aorta diameter. InPls3E10-16del/0rats,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 newPls3E10-16del/0rat 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 inPls3on the cardiovascular system need to be observed for a long time. This study expands the understanding of the cardiovascular phenotype of rarePls3-associated early-onset osteoporosis.更多还原