Growth effects of electrically stimulated contraction on adult feline cardiocytes in primary culture.
Growth effects of electrically stimulated contraction on adult feline cardiocytes in primary culture.
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电刺激收缩对原代培养中成年猫心肌细胞生长的影响。
DOI：
10.1152/ajpheart.1995.268.6.h2495
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发表时间：
1995
期刊：
影响因子：
--
通讯作者：
McDermott,PJ
中科院分区：
文献类型：
--
作者：
Kato,S;Ivester,CT;Cooper4th,G;Zile,MR;McDermott,PJ
本研究的目的是确定长期电刺激心肌收缩对蛋白质合成率和总蛋白质含量的影响。将成年猫心肌细胞镀在层粘胶蛋白包被的培养托盘上，并置于无血清培养基中，培养基由M199补充抗坏血酸盐、牛血清白蛋白、肌酸、肉碱、牛磺酸和10(-7)M重组胰岛素组成。使用频率为1赫兹、脉冲持续时间为5毫秒的交替极性连续电脉冲，电刺激心肌细胞收缩。未受刺激的心肌细胞通常处于静止状态，作为对照组。在对照组静止心肌中，蛋白质合成率在培养的第1 - 4天下降14%，然后在第7天保持稳定。在电刺激的心肌细胞中，蛋白质合成率在第1天和第7天增加了19%。在电刺激下，蛋白质合成率在第4天和第7天分别比静止心肌细胞高18%和43%。利用异硫氰酸荧光素染色细胞的共聚焦显微镜，通过测量每个细胞的总荧光来测定每个细胞的蛋白质含量。与对照组相比，电刺激7天后细胞蛋白含量显著增加52%。静息和电刺激的心肌细胞保持杆状，保留其肌纤维结构，并在7天内对电刺激有反应。这些数据表明，通过培养7天每个细胞蛋白质含量的增加，电刺激的成年心肌细胞收缩导致细胞生长。这种增长至少部分是由于稳态蛋白质合成速率的加速。
The purpose of this study was to determine effects of long-term electrical stimulation of cardiocyte contraction on protein synthesis rates and total protein content. Adult feline cardiocytes were plated on laminin-coated culture trays and maintained in a serum-free medium consisting of M199 supplemented with ascorbate, bovine serum albumin, creatine, carnitine, taurine, and 10(-7) M recombinant insulin. Cardiocytes were electrically stimulated to contract with use of continuous electrical pulses of alternating polarity at a frequency of 1 Hz and pulse duration of 5 ms. Nonstimulated cardiocytes are normally quiescent and were used as the control group. In control quiescent cardiocytes, protein synthesis rate decreased by 14% between days 1 and 4 in culture and then remained stable through day 7. In electrically stimulated cardiocytes, protein synthesis rates increased by 19% between days 1 and 7. Protein synthesis rates were 18% higher on day 4 and 43% higher on day 7 in electrically stimulated than in quiescent cardiocytes. Protein content per cell was determined by measuring total fluorescence per cell by use of confocal microscopy of fluorescein isothiocyanate-stained cells. Electrical stimulation significantly increased cellular protein content by 52% after 7 days compared with controls. Quiescent and electrically stimulated cardiocytes remained rod shaped, retained their myofibrillar architecture, and were responsive to electrical stimulation over the 7-day period. These data demonstrated that electrically stimulated contraction of adult cardiocytes resulted in cell growth, as assessed by an increase in protein content per cell over 7 days in culture. This increase was due, at least in part, to an acceleration of steady-state protein synthesis rates.