|本期目录/Table of Contents|

[1]尹琳 刘明鑫 王凤媛 唐艳红 王晞 赵庆彦 王腾 陈玉婷 黄从新.配对相关的同源异型框1导入棕色脂肪干细胞构建生物起搏[J].国际心血管病杂志,2019,06:351-357.
　YIN Lin,LIU Mingxin,WANG Fengyuan,et al.Paired-related homeobox 1 introduced into brown adipose-derived stem cells to trigger biological pacing[J].International Journal of Cardiovascular Disease,2019,06:351-357.
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配对相关的同源异型框1导入棕色脂肪干细胞构建生物起搏(PDF)

《国际心血管病杂志》[ISSN:1006-6977/CN:61-1281/TN]

期数:
2019年06期
页码:
351-357
栏目:
基础研究
出版日期:
2019-12-31

文章信息/Info

Title:
Paired-related homeobox 1 introduced into brown adipose-derived stem cells to trigger biological pacing
作者:
尹琳 刘明鑫 王凤媛 唐艳红 王晞 赵庆彦 王腾 陈玉婷 黄从新
430060 武汉大学人民医院心内科,武汉大学心血管病研究所,心血管病湖北省重点实验室
Author(s):
YIN Lin;  LIU Mingxin;  WANG Fengyuan;  TANG Yanhong;  WANG Xi;  ZHAO Qingyan;  WANG Teng;  CHEN Yuting;  HUANG Congxin
Department of Cardiology, Renmin Hospital of Wuhan University; Cardiovascular Research Institute, Wuhan University; Hubei Key Laboratory of Cardiology, Hubei 430060, China
关键词:
配对相关的同源异型框1;  生物起搏;  类窦房结细胞;  TBX18;  胰岛素基因增强子结合蛋白1
Keywords:
Paired related homeobox 1;  Biological pacing;  Sinus node-like cells;  TBX18;  ISL-1
分类号:
-
DOI:
10.3969/j.issn.1673-6583.2019.06.008
文献标识码:
-
摘要:
目的:探讨过表达配对相关的同源异型框1(Prrx1)是否能诱导棕色脂肪干细胞(BADSC)向类窦房结细胞分化,构建生物起搏。方法:分离培养大鼠BADSC,分别转染带有GFP的空载腺病毒(Ad-GFP组)和带有Prrx1的腺病毒(Ad-Prrx1组)。光镜下观察细胞形态和荧光表达强度,Western blot、实时聚合酶链反应检测窦房结细胞相关的起搏蛋白(HCN4)和转录因子[TBX18、胰岛素基因增强子结合蛋白1(ISL-1)、Pitx2]的表达水平,膜片钳技术检测起搏电流If,免疫荧光技术检测细胞HCN4、TBX18和ISL-1的表达。结果:Ad-Prrx1组的起搏相关因子TBX18、ISL-1、HCN4的mRNA和蛋白表达水平均明显高于Ad-GFP组,而Pitx2的mRNA表达水平明显降低(P均<0.05)。膜片钳记录到BADSC的If电流,且该电流能被4 mmol/L CsCl阻断。免疫荧光显微镜下可见Ad-Prrx1组Prrx1与TBX18、ISL-1、HCN4在BADSC中共表达,而Ad-GFP组未发现有上述起搏相关蛋白共表达。结论:过表达Prrx1能诱导BADSC分化为类窦房结细胞。
Abstract:
Objective:To investigate whether overexpression of paired-related homeobox 1(Prrx1)can successfully induce differentiation of brown adipose-derived stem cells(BADSCs)into sinus node-like cells.Methods:BADSCs were isolated and cultured, and the adenovirus with GFP(Ad-GFP group)and Prrx1(Ad-Prrx1 group)were transfected respectively. Light microscope was used to observe cell morphology and fluorescence expression intensity. Western blot and real-time polymerase chain reaction were used to detect the expression level of hyperpolarization-activated cyclic nucleotide-gated potassium channel 4(HCN4)and transcription factors related to sinus node cells(Tbx18, Isl-1and Pitx2). Whole-cell patch-clamp technique was used to record the pacing current hyperpolarization-activated inward current(If). Immunofluorescence technique was used to detect the expression of HCN4, Tbx18 and Isl-1.Results:The mRNA and protein expression levels of Tbx18, Isl-1 and HCN4 in Ad-Prrx1 group were significantly higher than those in Ad-GFP group, while the mRNA expression level of Pitx2 was significantly lower(P all<0.05). Whole-cell patch clamps were able to record the If current in Ad-Prrx1 group rather than in Ad-GFP group. The If current could be blocked by 4 mmol/L CsCl. Immunofluorescence analysis showed that Prrx1 was coexpressed with TBX18, ISL-1, and HCN4 in Ad-Prrx1 group, which did not appear in Ad-GFP group.Conclusions:Overexpression of prrx1 can induce BADSCs to differentiate into sinus node-like cells.

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备注/Memo

备注/Memo:
基金项目:湖北省技术创新专项(2016ACA153); 中央高校基本科研业务费专项资金(2042015kf0229)
作者单位:430060 武汉大学人民医院心内科,武汉大学心血管病研究所,心血管病湖北省重点实验室
通信作者:黄从新,Email:huangcongxin@vip.163.com
更新日期/Last Update: 2019-12-27