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目的:探讨心肌营养素-1(CT-1)在高血压心室重塑中的作用。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)、正义寡核苷酸对照组(SODN)、PD98059组、AG490组、LY294002组。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Western blotting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(0.132±0.013vs0.154±0.011,P<0.05),STAT3、ERK1/2和PI3-K蛋白表达水平明显低于对照组(2.09±0.25vs2.47±0.28,P<0.05)、(1.13±0.19vs1.61±0.22,P<0.05)、(1.25±0.23vs1.71±0.25,P<0.05)。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)并且增加ERK1蛋白磷酸化(1.85±0.18vs1.45±0.23,P<0.05);PD98059增强高静水压的促增殖(0.185±0.011vs0.155±0.010,P<0.05)并且增加STAT3蛋白磷酸化(1.83±0.23vs1.58±0.22,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);SOND(0.151±0.010vs0.154±0.011,P>0.05)和DMSO组(0.141±0.017vs0.155±0.010,P>0.05)与对照组相比对心肌成纤维细胞增殖无明显差别。结论:高静水压下,心肌成纤维细胞增殖主要通过STAT3通路;ERK1/2通路通过抑制STAT3的活性起负向调节作用,可能在防止心肌成纤维细胞过度增殖起重要的作用;PI3-K没有参与这一作用。上述STAT3通路与ERK1/2通路之间的相互作用可能有助于防止诱导成纤维细胞过度增殖。
Objective: To investigate the role of cardiotrophin-1 (CT-1) in hypertensive ventricular remodeling. Methods: The third-generation myofibroblasts were used in the experiment and divided into control group, DMSO (DMSO) group, ASODN CT-1 group, sense oligonucleotide Control group (SODN), PD98059 group, AG490 group, LY294002 group. Using self-made pressure culture device, each group of cells was placed under 160mmHg pressure for 8h. The activities of STAT3, ERK1 / 2 and PI3-K were determined by Western blotting; the proliferation of cardiac fibroblasts was measured by MTT assay. Results: CT-1ASODN could inhibit the proliferation of cardiomyocytes induced by high hydrostatic pressure (0.132 ± 0.013vs0.154 ± 0.011, P <0.05) after CT-1ASODN treatment. The protein expression of STAT3, ERK1 / 2 and PI3-K were significantly lower than those in the control group (2.09 ± 0.25 vs 2.47 ± 0.28, P <0.05) (1.13 ± 0.19 vs 1.61 ± 0.22, P <0.05) 0.23 vs 1.71 ± 0.25, P <0.05). AG490 group significantly attenuated the effects of high hydrostatic pressure on proliferation (0.118 ± 0.018vs0.155 ± 0.010, P <0.05) and increased phosphorylation of ERK1 (1.85 ± 0.18vs1.45 ± 0.23, P <0.05) (0.185 ± 0.011vs0.155 ± 0.010, P <0.05) and increased the phosphorylation of STAT3 (1.83 ± 0.23vs1.58 ± 0.22, P <0.05). After PI3 / K blocker LY294002 intervention, (0.157 ± 0.015vs0.155 ± 0.010, P> 0.05); SOND (0.151 ± 0.010vs0.154 ± 0.011, P> 0.05) and DMSO group (0.141 ± 0.017vs0.155 ± 0.010, P> 0.05) compared with the control group of myocardial fibroblasts proliferation no significant difference. CONCLUSION: Myocardial fibroblasts proliferate mainly through STAT3 pathway under high hydrostatic pressure. ERK1 / 2 pathway plays a negative regulatory role by inhibiting STAT3 activity, which may play an important role in preventing excessive proliferation of cardiac fibroblasts. PI3-K is not involved This role. The above interaction between the STAT3 pathway and the ERK1 / 2 pathway may help prevent the induced proliferation of fibroblasts.