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
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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