V_1受体拮抗剂对精氨酸升压素诱导的大鼠心脏成纤维细胞增殖的影响

Effects of V_1 Receptor Antagonist on the Proliferation of Rat Cardiac Fibroblasts Induced by Arginine Vasopressin

目的 探讨V1 受体拮抗剂 [d(CH2 ) 5 Tyr2 (Me) ]AVP对精氨酸升压素 (AVP)诱导的大鼠心脏成纤维细胞增殖的影响。方法 采用胰酶消化法培养新生Sprague Dawley (SD)大鼠心脏成纤维细胞 (CFs) ,以3H TdR掺入法测定CFs的DNA合成功能 ,MTT比色法测定CFs数目 ,并应用流式细胞仪进行CFs细胞周期分析。结果 ①CFs的3H TdR掺入率随着AVP干预浓度的增加而增高 ,其中 10 - 7mol LAVP和 10 - 6 mol LAVP组每 5 0 0 0个细胞的 3H TdR掺入率分别为(2 4 3± 6 1)cpm和 (32 8± 6 8)cpm ,均明显高于对照组3H TdR掺入率 (117± 32 )cpm(P <0 0 1) ;②MTT比色法吸光度 (A4 90nm)值随AVP浓度的增加而增高 ,其中 10 - 7mol LAVP、10 - 6 mol LAVP组的A4 90nm值分别为 0 2 4± 0 0 1和 0 2 9±0 0 2 ,均较对照组A4 90nm值 (0 16± 0 0 1)显著增高 (P <0 0 1) ;③ 10 7mol LAVP组CFs细胞周期S期百分率显著高于对照组 (30 2 0± 0 88) %vs(2 6 .86± 1 0 6 ) % (P <0 0 1) ;④ 10 - 7mol LAVP +10 - 7mol L [d(CH2 ) 5 Tyr2 (Me) ]AVP组的每5 0 0 0个细胞的3H TdR掺入率、MTT比色法A4 90nm值和S期百分率分别为 (14 3± 4 0 )cpm、0 17± 0 0 1和 (2 5 0 2±0 5 1) % ,均显著低于 10 - 7mol LAVP组 (分别P

Aim To investigate the effects of V 1 receptor antagonist [d(CH 2) 5 Tyr 2(Me)]AVP on the proliferation of rat cardiac fibroblasts (CFs) induced by arginine vasopressin (AVP). Methods CFs of neonatal SD rats were isolated by trypsin digestion method and growth arrested CFs were stimulated with 2.5% fetal calf serum in the presence of varying concentrations of AVP and 10 -7 mol/L[d(CH 2) 5 Tyr 2(Me)]AVP. DNA synthesis was measured by \{\}\+3H TdR incorporation. MTT and flowcytometry techniques were used to evaluate cell number and cell cycle respectively. Results ①AVP increased 3H TdR incorporation in cardiac fibroblasts in a concentration dependent manner. 3H TdR incorporation values in 10 -7 mol/L AVP and 10 -6 mol/L AVP group were(243±61)cpm/5 000 cells, (328±68)cpm/5 000 cells, respectively, which were significantly higher than that of the control (117±32) cpm/5000 cells(both P <0 01). The absorbance at 490 nM (A490 nM)was positively correlated with the concentration of AVP. A490 nm values in 10 -7 mol/L AVP and 10 -6 mol/L AVP group (0 24±0 01, 0 29±0 02, respectively) were significantly higher than that of the control (0 16±0 01, both P <0 01). Cells percentage in S stage in 10 -7 mol/L AVP group were significantly higher than that of the control (30 20±0 88)% vs (26.86±1 06)% ( P <0 01). ②In 10 -7 mol/L AVP+10 -7 mol/L [d(CH 2) 5 Tyr 2(Me)]AVP group, the 3H TdR incorporation in CFs, A490 nm value by MTT assay and S stage percentage were (143±40)cpm/5 000 cells, 0 17±0 01 and (25 02±0 51)%, which were significantly lower compared with 10 -7 mol/L AVP group( P <0 05, P <0 01, P <0 01, respectively). Conclusion The results indicate that AVP increases CFs number and DNA synthesis. The effects can be blocked by V 1 receptor antagonist [d(CH 2) 5 Tyr 2(Me)] AVP. These data suggest that AVP might promote CFs proliferation, which seems to be mediated via the V 1 receptor.