RESUMO
OBJECTIVE: The aim of this study was to investigate the role of microRNA-593-5p (miR-593-5p) in hypoxia-induced pulmonary hypertension (HPH), and to explore its underlying mechanism. MATERIALS AND METHODS: Sprague-Dawley (SD) rats were housed in a hypoxia environment 8 hours per day for consecutive 4 weeks. After the establishment of the HPH rat model, we detected the right ventricular systolic pressure (RVSP) and right heart hypertrophy index (RVHI) in HPH rats and controls. The expression levels of miR-593-5p and polo-like kinase 1 (PLK1) in rat lungs were detected by quantitative Real Time-Polymerase Chain Reaction (qRT-PCR). Subsequently, miR-593-5p mimics and inhibitor were constructed and transfected into cells. The proliferation and migration of pulmonary arterial smooth muscle cells (PASMCs) were accessed by cell counting kit-8 (CCK-8) assay and wound healing assay, respectively. The protein level of PLK1 in PASMCs after transfection with miR-593-5p mimics or inhibitor was detected by Western blot. Dual-luciferase reporter gene assay was conducted to verify the binding condition of miR-593-5p and PLK1. Finally, rescue experiments were performed to explore whether the regulatory effect of miR-593-5p on HPH development was associated with PLK1. RESULTS: RVSP and RVHI in rats of the hypoxic group were significantly higher than those of controls. MiR-593-5p was significantly downregulated while PLK1 was remarkably upregulated in lung tissues of HPH rats than those of controls. Similarly, miR-593-5p expression in PASMCs decreased gradually with the prolongation of hypoxia induction. Overexpression of miR-593-5p remarkably inhibited the proliferation and migration of PASMCs. Subsequently, dual-luciferase reporter gene verified the binding condition of miR-593-5p and PLK1. Both the mRNA and protein levels of PLK1 were negatively regulated by miR-593-5p. Also, rescue experiments demonstrated that the inhibitory effects of miR-593-5p on the proliferation and migration of PASMCs could be reversed by PLK1 overexpression. CONCLUSIONS: MiR-593-5p is lowly expressed in lung tissues of HPH rats. Meanwhile, it stimulates the proliferation and migration of PASMCs via targeting PLK1, thereby promoting HPH development.
Assuntos
Proteínas de Ciclo Celular/genética , Hipertensão Pulmonar/genética , Hipóxia/complicações , MicroRNAs/metabolismo , Proteínas Serina-Treonina Quinases/genética , Proteínas Proto-Oncogênicas/genética , Remodelação Vascular/genética , Animais , Hipóxia Celular , Linhagem Celular , Movimento Celular/genética , Proliferação de Células/genética , Modelos Animais de Doenças , Regulação para Baixo , Humanos , Hipertensão Pulmonar/patologia , Pulmão/irrigação sanguínea , Pulmão/patologia , Masculino , Músculo Liso Vascular/citologia , Músculo Liso Vascular/patologia , Miócitos de Músculo Liso/patologia , Cultura Primária de Células , Artéria Pulmonar/citologia , Artéria Pulmonar/patologia , Ratos , Ratos Sprague-Dawley , Regulação para Cima , Quinase 1 Polo-LikeRESUMO
In addition to the known cycleanine (I) and d-isochondodendrine (II), two novel bisbenzylisoquinoline alkaloids named isocycleanine (III) and sutchuenensine (IV) have been isolated from the roots of Cyclea sutchuenensis Gagnep grown wild in Sichuan province, China. III was established as the epimer of I and IV, as a new type of head-to-tail bisbenzylisoquinoline alkaloid with two diarylether bridges in between C-8 to C-12' and C-12 to C-7' on the basis of physical constants and spectral data (UV, IR, EIMS, 1HNMR and NOEDS).
Assuntos
Alcaloides/isolamento & purificação , Medicamentos de Ervas Chinesas/química , Isoquinolinas , Alcaloides/química , Estrutura MolecularRESUMO
Oxidation of insularine (I) with m-chloroperbenzoic acid yielded four insularine-N-oxides. Two of them are identical in every respect with our insularine-2 beta-N-oxide (II) and insularine-2' beta-N-oxide (III), two rare examples of naturally occurring head-to-tail bisbenzylisoquinoline N-oxides newly isolated from Cyclea sutchuenensis, which confirmed the structures of the two novel natural N-oxides. The other two are new compounds and their structures have been established as insularine-2' alpha-N-oxide (IV) and insularine-2 beta,2' beta-N, N-dioxide (V), on the basis of spectral data (UV, IR, 1HNMR, NOEDS AND MS) analysis.