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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。
AIM: To investigate the effects of arginine vasopressin (AVP) and its V1a receptor on the myofibroblasts (MFs) transdifferentiation in cultured SD SD rats. Methods: CFs were isolated and cultured from SD rats by trypsin digestion. CFs were incubated with AVP of different concentrations (10-9, 10-8, 10-7, 10-6 mmol / L) AVP V1a-specific receptor antagonist [d (CH2) 5Tyr2 (Me)] AVP was incubated with 10-6 mmol / L AVP for 48 h. Proline incorporation assay was used to detect collagen synthesis in CFs. Western blot was used to detect the expression of α-SMA in CFs. CFs morphology was observed by α-SMA immunofluorescence staining. Results: The concentration of 3 H-proline incorporation and α-SMA in CFs induced by AVP were increased in a concentration-dependent manner. The incorporation of 3 H-proline in CFs of 10-6 mmol / L AVP group and α- (P <0.05). The volume of CFs increased significantly under the action of 10-6 mmol / L AVP, and the fluorescence intensity of cytoplasm was significantly enhanced. The structure of cytoplasm showed obvious tension-like structure . However, [d (CH2) 5Tyr2 (Me)] AVP could inhibit the increase of 3H-proline incorporation and α-SMA expression in CFs induced by 10-6 mmol / L AVP in a concentration-dependent manner. , The 3H-proline incorporation of CFs in 10-7 mmol / L [d (CH2) 5Tyr2 (Me)] AVP group was significantly lower than that in 10-6 mmol / L AVP group (all P <0.05) The expression of α-SMA of CFs in -9,10-8,10-7 mmol / L [d (CH2) 5Tyr2 (Me)] AVP group was significantly lower than that in 10-6 mmol / L AVP group (all P <0.05) , And 10-7 mmol / L [d (CH2) 5Tyr2 (Me)] AVP could induce 3 H-proline incorporation, α-SMA expression and morphology of CFs induced by 10-6 mmol / L AVP Changes were inhibited to the control group (all P> 0.05). Conclusions: AVP induces the conversion of CFs to MFs via its V1a receptor.