增龄对大鼠心成纤维细胞生长和功能的影响  

Effects of aging on the growth and function of cardiac fibroblasts in rats

作　　者：王琰 许亚平 王前 罗欢欢 赵育洁 牛思泉 胥良[1] 郭志坤 Wang Yan;Xu Yaping;Wang Qian;Luo Huanhuan;Zhao Yujie;Niu Siquan;Xu Liang;Guo Zhikun(Department of Cardiology,The Seventh People’s Hospital of Zhengzhou,Zhengzhou 450006;Zhengzhou Key Laboratory of Heart Structure Research,The Seventh People's Hospital of Zhengzhou,Zhengzhou 450006;School of Nursing,Xinxiang College,Xinxiang 453003;Henan Key Laboratory of Medical Tissue Regeneration,Xinxiang Medical University,Xinxiang 453003,China)

机构地区：[1]郑州市第七人民医院心内科,郑州450006 [2]郑州市第七人民医院郑州市心脏结构研究重点实验室,郑州450006 [3]新乡学院护理学院,新乡453003 [4]新乡医学院河南省医用组织再生重点实验室,新乡453003

出　　处：《解剖学杂志》2022年第1期1-5,F0002,共6页Chinese Journal of Anatomy

基　　金：国家自然科学基金(81570268);河南省卫生厅攻关项目(15210231011);河南省自然科学基金青年项目(212300410305);郑州市心脏结构研究重点实验室资助项目(002);河南省医学科技攻关计划联合共建项目(LHGJ20191115)。

摘　　要：目的:探讨增龄对大鼠心成纤维细胞(CFs)生长和功能的影响。方法:体外培养新生、1月龄、3月龄大鼠CFs,利用MTT法检测细胞生长能力变化;免疫荧光、免疫细胞化学检测干细胞表面标志物nanog、sox2、oct4、scal-1和c-kit的表达差异,利用成脂、成骨、成心肌诱导液检测各组CFs多向分化能力差异。结果:培养的各组CFs均表达盘状结构域受体2(DDR2),符合心成纤维细胞特性;贴壁2~4d时,各组CFs处于对数生长期,4~5 d时处于平台期,5~6 d时处于下降期;贴壁2~6 d时,新生组CFs生长速度高于1月龄组,1月龄组CFs生长速度高于3月龄组;各组CFs均表达nanog、sox2、oct4、scal-1、c-kit,nanog的荧光定量结果为新生组>3月龄组>1月龄组,sox2的荧光定量结果为新生组>1月龄组>3月龄组,oct4的荧光定量结果为新生组>1月龄组>3月龄组,scal-1的荧光定量结果为1月龄组>新生组>3月龄组,c-kit的荧光定量结果为1月龄组和3月龄组>新生组,而1月龄组和3月龄组间差异无统计学意义。各组CFs在多向诱导分化后,均有成脂、成骨、成心肌能力。结论:增龄可影响CFs的生长速度和多种表面标志物表达量,但对其多向分化潜能并无明显影响。Objective:To investigate the effect of aging on the growth and function of cardiac fibroblasts(CFs)in rats.Methods:CFs of neonatal,1-month-old,and 3-month-old rats were cultured in vitro.MTT assay was used to detect the change of cell growth ability.Immunofluorescence and immunocytochemistry were used to detect the expression of nanog,sox2,oct4,scal-1,and c-kit,and the differentiation ability of each group was detected by adipogenic,osteogenic and cardiogenic inducers.Results:The expression of discoindin domain receptor 2(DDR2)in CFs of each group was consistent with the characteristics of cardiac fibroblasts.The CFs of each group were in logarithmic growth stage at 2-4 d,plateau stage at 4-5 d,and descending phase at 5-6 d.The growth rate of CFs in the neonatal group were higher than that in the 1-month-old group,and was higher in the 1-month-old than that in the 3-month-old group at 2-6 d.CFs of each group expressed nanog,sox2,oct4,scal-1,and c-kit.The fluorescence quantitative results of nanog were as follows:newborn group>3-month-old group>1-month-old group.Sox2 was newborn group>1-month-old group>3-month-old group.Oct4 was newborn group>1-month-old group>3-month-old group.Scal-1 was 1-month-old group>newborn group>3-month-old group.c-Kit was 1-month-old group and 3-month-old group>newborn group,but there was no significant difference between 1-month-old group and 3-month-old group.CFs of each group had the ability of adipogenic,osteogenic,and myocardial formation after multi-directional differentiation induction.Conclusion:Aging can affect the growth rate of CFs and their expression levels of various surface markers,but has no significant effect on the multi-directional differentiation potential of CFs.

关 键 词：心成纤维细胞 增龄 诱导分化 表面标志物 大鼠 

分 类 号：R329.2[医药卫生—人体解剖和组织胚胎学]