摘要
目的:探讨精氨酸加压素(AVP)及其V1a受体对体外培养的SD仔鼠心肌成纤维细胞(CFs)向肌成纤维细胞(MFs)转化的影响。方法:胰酶消化法分离培养SD仔鼠CFs,将CFs分别与不同浓度(10-9、10-8、10-7、10-6 mmol/L)AVP,或添加了不同浓度(0、10-9、10-8、10-7 mmol/L)AVP V1a特异性受体拮抗剂[d(CH2)5Tyr2(Me)]AVP的10-6 mmol/L AVP共同孵育48h后,用3 H-脯氨酸掺入法检测CFs胶原合成功能,Western blot检测CFs平滑肌肌动蛋白α(α-SMA)表达量,α-SMA免疫荧光染色观察CFs形态。结果:AVP浓度依赖性地诱导CFs的3 H-脯氨酸掺入率和α-SMA表达量增加,其中10-6 mmol/L AVP组CFs的3 H-脯氨酸掺入率和α-SMA表达量均显著高于对照组(均P<0.05),且10-6 mmol/L AVP作用下CFs体积明显增大,胞质荧光染色亮度显著增强,胞质中有明显的张力丝样结构。而[d(CH2)5Tyr2(Me)]AVP可以浓度依赖性地抑制10-6 mmol/L AVP诱导下CFs 3 H-脯氨酸掺入率和α-SMA表达量的增加,其中10-8、10-7 mmol/L[d(CH2)5Tyr2(Me)]AVP组中CFs的3 H-脯氨酸掺入率显著低于10-6 mmol/L AVP组(均P<0.05),10-9、10-8、10-7 mmol/L[d(CH2)5Tyr2(Me)]AVP组中CFs的α-SMA表达量显著低于10-6 mmol/L AVP组(均P<0.05),且10-7 mmol/L[d(CH2)5Tyr2(Me)]AVP能够将10-6 mmol/L AVP诱导下CFs的3 H-脯氨酸掺入率、α-SMA表达量和形态学变化都抑制到对照组水平(均P>0.05)。结论:AVP通过其V1a受体诱导CFs向MFs转化。
Objective：To investigate the effects of arginine vasopressin （AVP） and its Vla receptor on differentiation of cardiac fibroblasts （CFs） into myofibroblasts （MFs）. Method： CFs was isolated by trypsin digestion method, and was co-incubated for 48 h with different concentrations （10 ^-9, 10^-8, 10^-7, 10^-6 mmol/L） of AVP, or 10 ^-6 mmol/L AVP supplemented with different concentrations （0, 10^-9 , 10^-8 , 10^-7 mmol/L） of Vla receptor selective antagonist [d（CH2）5Tyr2（Me）]AVP, respectively. 3 H proline incorporation rate was used to measure collagen synthesis, Western blot analysis was used to measure smooth muscle actin a （a-SMA） expression, and immunofluorescent staining for a-SMA was used to detect morphological changes of CFs. Result： AVP dosedependently increased the s H proline incorporation rate and a-SMA expression in CFs. 10^-6 mmol/L AVP significantly increased 3H proline incorporation rate and a-SMA expression compared with control group （P〈0.05）. CFs stimulated by 10^-6 mmol/L AVP showed larger cell size, brighter staining for a-SMA and appearance of stress fibers. [d（CH2）5Tyr2 （Me）]AVP dose-dependently inhibited the increases of 3H proline incorporation rate and a-SMA expression induced by 10^- 6 mmol/L AVP in CFs （P〈0.05）. 10 ^-7 mmol/L [d（CH2）5Tyr2（Me）] AVP down-regulated the levels of 3 H proline incorporation rate, a-SMA expression and morphological changes of CFs induced by 10^-6 mmol/L AVP to baseline （P〉0. 05）. Conclusion： AVP can induce the differentiation of CFs into MFs via its Vla receptor.
出处
《临床心血管病杂志》
CAS
CSCD
北大核心
2013年第2期152-154,共3页
Journal of Clinical Cardiology
