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目的:探讨肾上腺髓质素N端20肽(PAMP)对血管紧张素Ⅱ(angotesinⅡ,AngⅡ)刺激心肌成纤维细胞(CFs)增殖及胶原生成的影响。方法:采用胰酶消化、差速贴壁法培养新生SD大鼠CFs,以3H脯氨酸掺入法测定胶原合成、四氮唑盐(MTT)比色法测定细胞数目,分别观察不同浓度的PAMP、AngⅡ、及PAMP对ANGⅡ诱导CFs增生及胶原合成作用的影响。实验分组(1)空白对照组;(2)10-9、10-8、10-7、10-6mol/LAngⅡ组;(3)10-9、10-8、10-7、10-6mol/LPAMP组;(4)10-7mol/LAngⅡ+PAMP(10-9、10-8、10-7、10-6mol/L)组。结果:(1)随着PAMP浓度的增高,MTT比色值差异无显著性(F=10.21,P>0.05)。随着AngⅡ浓度的增高,MTT比色值也明显增高,各组间差异有显著性(P值均<0.01)。PAMP+AngⅡ组,随着PAMP浓度的增高,MTT比色值均显著降低(P<0.01)。(2)PAMP组的3H脯氨酸掺入率与对照组相比(F=16.92,P>0.05),各组之间差异无显著性。随着AngⅡ浓度的增高,CFs的3H脯氨酸掺入率呈递增趋势。PAMP+AngⅡ组的3H脯氨酸掺入率在AngⅡ浓度不变的情况下,随着PAMP浓度的增高,CFs的3H脯氨酸掺入率呈递减趋势,各组之间差异有显著意义(P<0.05)。结论:PAMP对AngⅡ刺激心肌成纤维细胞增生及胶原的生成起明显的抑制作用。
Objective: To investigate the effect of adrenomedullin N-terminal 20 peptide (PAMP) on the proliferation and collagen production of angiotensin Ⅱ (Ang Ⅱ) stimulated cardiac fibroblasts (CFs). Methods: CFs of newborn SD rats were cultured by trypsin digestion and differential adherent method. Collagen synthesis was measured by 3H proline incorporation method and cell number was determined by MTT assay. Effects of PAMP, Ang Ⅱ and PAMP on ANG Ⅱ - induced CFs Proliferation and Collagen Synthesis. (2) 10-9,10-8,10-7,10-6mol / LAngⅡgroup; (3) 10-9,10-8,10-7,10-6 mol / LPAMP group; (4) 10-7mol / LAngⅡ + PAMP group (10-9,10-8,10-7,10-6mol / L). Results: (1) With the increase of PAMP concentration, MTT colorimetric values showed no significant difference (F = 10.21, P> 0.05). With the increase of Ang Ⅱ concentration, MTT colorimetric values also increased significantly, the difference between the groups was significant (P value <0.01). PAMP + Ang Ⅱ group, with the increase of PAMP concentration, MTT colorimetric values were significantly reduced (P <0.01). (2) 3H-proline incorporation in PAMP group was significantly higher than that in control group (F = 16.92, P> 0.05). There was no significant difference between groups. With the increase of Ang Ⅱ concentration, 3H proline incorporation rate of CFs showed an increasing trend. The incorporation rate of 3H proline in PAMP + AngⅡ group decreased with the increase of PAMP concentration at the same concentration of AngⅡ, and the incorporation rate of 3H proline in CFs tended to decrease (P < P <0.05). CONCLUSION: PAMP significantly inhibits the proliferation of cardiac fibroblasts stimulated by AngⅡ and collagen production.