肾上腺髓质素N端20肽对血管紧张素Ⅱ刺激心肌成纤维细胞增殖、胶原合成的影响

The inhibitory effects of PAMP on the proliferation and collagen synthesis in rat cardiac fibroblasts induced by AngⅡ

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 effects of proADM N-terminal 20 peptide （PAMP） on the proliferation and collagen synthesis of rat cardiac fibroblasts （CFs） induced by Angiotensin Ⅱ （AngⅡ）. Methods CFs of neonatal Sprague-Dawley （SD） rats were isolated with trypsin digestion method and those growth arrested were stimulated with various concentrations of PAMP, various concentrations of AngⅡ and 10-Tmol/L AngⅡ in the presence of various concentrations of PAMP. The collagen synthesis of CFs was measured with ^3H （^3H） incorporation. MTr colorimetry was adopted to evaluate cell number. Group： ①blank group; ②10^-9, 10^-8, 10^-7, 10^-6mol/L AngⅡ groups; ③10^-9, 10^-8, 10^-7, 10^-6mol/L PAMP groups; ④10^-7mol/L AngⅡ＋PAMP（10^-9, 10^-8, 10^-7, 10^-6 mol/L） groups. Results ① A490 values of PAMP groups were not significantly changed（F=10.21, P〉0.05）; AngⅡ increase A490 values in CFs in a concentration dependent manner（P〈0.01）. A490 values of AngⅡ＋PAMP groups were significantly lower in comparison with 10^-7mol/L AngⅡ group （P〈0.01）. ②^3H incorporation values of PAMP groups were not significantly changed （F=16.92, P〉0.05）; Ang Ⅱ increase ^3H incorporation in CFs in a concentration dependent manner（P〈0.01）. ^3H incorporation in CFs value of AngⅡ＋PAMP groups was significantly lower in comparison with 10^-7mol/ L AngⅡ group （P〈0.01）. Conclusions It is suggested that PAMP can decrease CFs number and collagen synthesis induce by AngⅡ and may play an important role in the CFs proliferation.