Exosomal miR-211 contributes to pulmonary hypertension via attenuating CaMK1/PPAR-γaxis
Autor: | Linbo Yuan, Shu-Hao Zhang, Yiruo Sun, Jiantao Liu, Yufan Lin, Zhengze Yao, Kexin Lin, Luowei Chen, Kaidi Zheng, Yupeng Sun |
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Rok vydání: | 2020 |
Předmět: |
0301 basic medicine
Male medicine.medical_specialty Physiology Myocytes Smooth Muscle Peroxisome proliferator-activated receptor 030204 cardiovascular system & hematology Pulmonary Artery Vascular Remodeling Exosomes Exosome Muscle Smooth Vascular Rats Sprague-Dawley 03 medical and health sciences 0302 clinical medicine Downregulation and upregulation Western blot Right ventricular hypertrophy Internal medicine medicine Animals Hypoxia Cells Cultured Cell Proliferation Pharmacology chemistry.chemical_classification Pulmonary Arterial Hypertension Lung medicine.diagnostic_test Chemistry Hypoxia (medical) medicine.disease Pulmonary hypertension PPAR gamma Disease Models Animal MicroRNAs 030104 developmental biology medicine.anatomical_structure Endocrinology Calcium-Calmodulin-Dependent Protein Kinase Type 1 Molecular Medicine medicine.symptom Signal Transduction |
Zdroj: | Vascular pharmacology. 136 |
ISSN: | 1879-3649 |
Popis: | Aim Exsomes play a significant role in increasing pathophysiological processes by delivering their content. Recently, a variety of studies have showed exosomal microRNAs (miRNAs) are involved in pulmonary hypertension (PH) notably. In this study, we found that exosomal miR-211 was overexpressed in hypoxia-induced PH rats but its intrinsic regulation was unclear. Therefore, our aim was to reveal the underlying mechanism which overexpressed exosomal miR-211 targeted in the development of PH. Methods 18 male SD rats were randomly divided into normoxia and hypoxia group, housed in normal or hypoxic chamber for 3 weeks respectively. Then, mean pulmonary arterial pressure (mPAP), pulmonary vascular resistance(PVR), right ventricular hypertrophy index(RV/(LV + S)), the percentage of medial wall area (WA%) and the percentage of medial wall thickness (WT%) were measured. Expression of miR-211 in exosomes was detected by qRT-PCR. Expression of Ca2+/calmodulin-dependent kinase1(CaMK1)and peroxisome proliferator-activated receptors-γ(PPAR-γ)in lung tissue were detected by Western blot(WB); After miR-211 overexpressed exosomes were injected to rats through caudal vein, mPAP, PVR, RV/(LV + S), WA% and WT% were also measured. Sequentially, hypoxia rats were injected with lentivirus riched in miR-211 inhibitor via tail vein, and PH-related indicators were measured. In vitro, after miR-211 was positively or negatively regulated in pulmonary arterial smooth muscle cell (PASMC) by plasmid transfection, proliferation of PASMC was detected by CCK8, as well as the expression of CaMK1 and PPAR- γ. Further, the relationship between CaMK1 and miR-211 was verified by Dual-Luciferase assay. And the regulatory relationship of CaMK1/PPAR- γ aixs was demonstrated in PASMC. Results Evident increases of mPAP, PVR, RVHI, WT% and WA% were observed with hypoxia administration. And the concentration of plasma exosomes in hypoxia rats was increased and positively correlated with the above indexes. miR-211 in exosomes of PH was upregulated while the expression of CaMK1 and PPAR-γ decreased in lung tissues. Further, injection of exosomes overexpressed with miR-211 demonstrated that exosomal miR-211 aggravated PH while inhibition of miR-211 attenuated PH in rats. In vitro, overexpression of miR-211 promoted the proliferation of PASMC and inhibited expression of CaMK1 and PPAR-γ in PASMC. And Dual-luciferase assay demonstrated that CaMK1 was a downstream gene of miR-211. Plasmid transfection experiments indicated that CaMK1 can promote PPAR-γ expression. Conclusion Exosomal miR-211 promoted PH via inhibiting CaMK1/PPAR-γ axis, promoting PASMC proliferation in rats. |
Databáze: | OpenAIRE |
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