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糜酶对大鼠心脏成纤维细胞增殖和胶原合成的影响(PDF)

《心脏杂志》[ISSN:1009-7236/CN:61-1268/R]

期数:
2008年第3期
页码:
268-272
栏目:
基础研究
出版日期:
2008-05-20

文章信息/Info

Title:
Effects of chymase on proliferation and collagen synthesis of rat cardiac fibroblasts
作者:
赵晓燕; 赵连友; 郑强荪; 何燕萍; 陆小龙
第四军医大学唐都医院心内科，陕西 西安 710038
Author(s):
ZHAO Xiaoyan;  ZHAO Lianyou;  ZHENG Qiangsun;  HE Yanping;  LU Xiaolong
Department of Cardiology, Tangdu Hospital, Fourth Military Medical University, Xi’an 710038, Shaanxi, China
关键词:
糜酶; 心脏成纤维细胞; 增殖; 胶原; 大鼠
Keywords:
chymase;  cardiac fibroblast;  proliferation;  collagen;  rat
分类号:
R977.3
DOI:
-
文献标识码:
A
摘要:
目的 探讨糜酶对大鼠心脏成纤维细胞（CFs）增殖和胶原合成的影响。方法 用胰酶消化法分离、培养新生SD大鼠的CFs，采用MTT比色法（A490值）测定细胞数目，流式细胞术分析细胞周期，3H脯氨酸掺入法测定总胶原合成， RTPCR检测Ⅰ型和 Ⅲ型胶原mRNA的表达。结果 ①糜酶以剂量依赖方式增加CFs的数目。其中15、30 和 60 μg/L组的A490值分别为0263±0033、0348±0031和0387±0026，均高于对照组的A490值（0201±0019），并有非常显著性差异（P<001)。②随着糜酶作用浓度的增加，CFs在G0/G1期的百分率逐渐减少，S期的百分率和细胞增殖指数逐渐增加。其中15、30 和 60 μg/L组与对照组相比，上述各项指标均有显著或非常显著性差异（P<005或P<001）。③糜酶以剂量依赖方式增加CFs的3H脯氨酸掺入量。其中15、30和60 μg/L组的3H脯氨酸掺入量分别为（520±75）、（684±62）和（769±58）cpm/孔，均高于对照组\[(435±60)cpm/孔\]，差异有显著或非常显著性意义（P<005 或P<001）。④随着糜酶作用浓度的增加，CFs的Ⅰ型和 Ⅲ 型胶原mRNA表达水平均呈递增趋势。其中15、30和60 μg/L组的Ⅰ、Ⅲ型胶原mRNA表达的水平均显著高于对照组（P<001）。结论 糜酶具有促进CFs增殖和胶原合成的作用，提示其可能在心肌纤维化过程中发挥重要作用。
Abstract:
AIM To investigate the effects of chymase on the proliferation and collagen synthesis of rat cardiac fibroblasts (CFs). METHODS Cultured CFs from neonatal SD rats were isolated by trypsinization and cellular number and cellular cycle were evaluated by MTT colorimetry (A490 value) and flow cytometry (FCM). The total collagen synthesis by CFs was examined by 3Hproline incorporation and the expressions of TypesⅠ and Ⅲ collagen mRNA in CFs were determined by RTPCR. RESULTS ①Chymase increased CFs number in a dosedependent manner. The A490 values of 15, 30 and 60 μg/L chymase groups were 0263±0033, 0348±0031 and 0387±0026, respectively, significantly higher than those in control group（0201±0019, P<001). ②Cellular cycle analysis showed that treatment with 15, 30 and 60 μg/L chymase markedly decreased cellular percentage in G0/G1 stage and increased cellular percentage in S stage and proliferation index (PI) in a dosedependent manner compared with those in control groups (P<005 or P<001). ③Chymase increased the 3Hproline incorporation in CFs in a dosedependent manner. The 3Hproline incorporations induced by 15, 30 and 60 μg/L chymase were (520±75), (684±62) and (769±58)cpm/well, respectively, significantly higher than those in control group \[(435±60)cpm/well, P<005 or P<001\]. ④15, 30 and 60 μg/L chymase obviously increased the expressions of TypesⅠ and Ⅲ collagen mRNA in a dosedependent manner compared with those in control groups (P<001). CONCLUSION Chymase promotes the proliferation and collagen synthesis of rat CFs, which is involved in the process of myocardial fibrosis.

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备注/Memo

备注/Memo:
收稿日期：2008-01-02.基金项目：陕西省科学技术研究发展计划项目资助（2005K13G13） 通讯作者：赵连友,教授,主任医师,主要从事高血压发病机制及防治的研究Email:zhaolyfmmu@yahoo.com.cn 作者简介：赵晓燕,副主任医师,博士生Email:zxysxy420@yahoo.com.cn
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