摘要
目的探讨在高静水压条件下心肌营养素1(CT-1)对成纤维细胞增殖作用机制。方法3～4代心肌成纤维细胞用于实验,分为对照组、助溶剂(二甲亚砜,DMSO)组、CT-1反义寡核苷酸组(ASODN)、正义寡核苷酸组(SODN)、MEK-EPK阻断剂PD98059组、JAK-STAT3阻断剂AG490组、PI3K阻断剂LY294002。利用自制压力培养装置,将各组细胞置于160mmHg压力下培养8h。STAT3、ERK1/2和PI3-K的活性通过Westernblot分析测定;MTT测定心肌成纤维细胞增殖。结果高静水压明显促进心肌成纤维细胞增殖,CT-1表达上调,CT-1ASODN干预后,CT-1ASODN能抑制高静水压所致的细胞增殖,吸光度值(A值)为(0.132±0.013vs0.154±0.011,P<0.05),STAT3(2.09±0.25vs2.47±0.28,P<0.05)、ERK1/2(1.13±0.19vs1.61±0.22,P<0.05)和PI3-K(1.25±0.23vs1.71±0.25,P<0.05),蛋白表达水平明显低于对照组。AG490组明显减弱高静水压的促增殖作用(0.118±0.018vs0.155±0.010,P<0.05);PD98059增强高静水压的促增殖(0.185±0.011vs0.155±0.010,P<0.05),PI3-K阻断剂LY294002干预后对高静水压的促增殖作用无影响(0.157±0.015vs0.155±0.010,P>0.05);SODN与对照组相比对心肌成纤维细胞增殖无明显影响。结论高静水压下,可以激活STAT3、ERK1/2、PI3-K信号通路,心肌成纤维细胞增殖主要通过STAT3通路;ERK1/2通路起负向调节作用;PI3-K与细胞增殖关系不是很密切。
Objective To investigate the mechanism of cardiotrophin-1（CT-1） induced proliferation of cardiac fibroblasts under high hydrostatic pressure. Methods Cardiac fibroblasts were cultured in vitro, and subjected to different intervention factors： group C （control） ; group P： （cultured under high hydrostatic pressure 160 mm Hg）; group ASODN： （interfered with CT-1 anti-sense oligodeoxynucleotides 10 μmol/L）; group SODN： （incubated with CT-1 sense oligodeoxynucleotides 10 μmol/L）. Group AG： （with AG490 25 μmol/L）, group PD： （with PD98059 20 μmol/L）, group LY： （with LY 294002 10 μmol/L）. Western-blotting were employed to assess the expression of STAT3, ERK1/2 and PI3-K at protein level. Cell proliferation was quantified by MTT. Results High hydrostatic pressure stimulated the proliferation of cardiac fibroblasts, upregulated the expression of CT-1, CT-1 ASODN inhibited the proliferation of cardiac fibroblasts （ 0. 132 ± 0. 013 vs 0. 154 ± 0.011, P 0. 05）, ASODN inhibited the expression of STAT3, ERK1/2 and PI3-K respectively at protein level（2.09±0. 25 vs 2. 47 ±0. 28, P〈0. 05）, （1.13±0. 19 vs 1.61±0.22, P〈0.05）, （1.25±0.23 vs 1.71±0.25, P〈0.05）. AG490, an JAK-STAT3 inhibitor, reversed the increase of the proliferation of cardiac fibroblasts stimulated by high hydrostatic pressure. PD98059, an MAPK-ERK1/2 inhibitor, increased the proliferation of cardiac fibroblasts stimulated by high hydrostatic pressure C0. 185±0. 011 vs 0. 155±0. 010, P〈0. 05）. LY294002, an PI3-Kinhibitor, had no effect on the proliferation of cardiac fibroblasts stimulated by high hydrostatic pressure（0. 157 ±0. 015 vs 0. 155±0. 010, P〉0.05）. While no differences of the expression of the above factors were found in SODN（0. 151±0.010 vs 0. 154±0.011, P〉0.05） and DMSO（0. 141±0.017 vs 0. 155±0.010, P〉0.05） as compared with the control group. Conclusion High hydrostatic pressure can active STAT3, ERK1/2, PI3-K, the proliferation of cardiac fibro
出处
《中华高血压杂志》
CAS
CSCD
北大核心
2007年第3期228-232,共5页
Chinese Journal of Hypertension
基金
江西省科学技术厅重大项目
关键词
心肌营养素1
心肌成纤维细胞
心室重塑
信号转导
Cardiotrophin-1
Cardiac fibroblasts
Ventricular remodeling
Signal transduction
