ABSTRACT
This paper explores the potential mechanism of microRNA-143-5p regulation effects on pulmonary artery smooth muscle cells (PASMCs) functions in hypoxic pulmonary hypertension (HPH) via targeting HIF-1a, which may offer a new idea for HPH therapy. PASMCs were transfected with mimics control/miR-143-5p mimics or inhibitor control/miR-143-5p inhibitor. We used Western blotting and RT-qPCR to detect the protein and mRNA expressions, CCK-8 assay to detect cellular viability, Annexin V-FITC/PI staining and caspase- 3/cleaved caspase-3 protein to evaluate cellular apoptosis, transwell migration experiment for cellular migration measurement and Dual luciferase reporter gene assay to prove the target of miR-143-5p. Cells under hypoxic condition presented the decreased protein and mRNA expressions of α-smooth muscle actin (SM-α-actin), Myocardin, smooth muscle myosin heavy chain (SMMHC), and smooth muscle-22α (SM22α), Calponin1 and Hypoxia-inducible factor-1α(HIF-1α), the increased cell viability and miR-143-5p level; Overexpression of miR-143-5p obviously reduced vascular smooth muscle-specific contraction marker protein levels and cellular apoptosis, increased cellular migration of PASMCs with hypoxia stimulation; Low-expression of miR-143-5p caused the opposite changes, while co-transfected with Si HIF-1 α blocked the beneficial effects of miR-143-5p inhibition on PASMCs under hypoxia. MicroRNA-143-5p can promote the phenotype conversion, proliferation and migration of pulmonary artery smooth muscle cells under hypoxic condition through direct targeting of HIF-1α.
Subject(s)
Hypoxia-Inducible Factor 1, alpha Subunit/metabolism , MicroRNAs/metabolism , Muscle, Smooth, Vascular , Myocytes, Smooth Muscle/physiology , Oxygen/pharmacology , Pulmonary Artery , Cell Migration Assays , Cells, Cultured , Gene Expression Regulation/drug effects , Gene Expression Regulation/physiology , Humans , Hypoxia-Inducible Factor 1, alpha Subunit/genetics , MicroRNAs/geneticsABSTRACT
OBJECTIVE: To explore the influences of different dosage ferrous sulfate supplements on bone marrow hemopoiesis in rats. METHODS: Female weaning Wistar rats were fed with an iron deficient diet (< 10 mg/kg diet) until the level of hemoglobin of rats was lower than 100 g/L. Rats (n = 50) were randomly divided into five groups according to the levels of hemoglobin and body weight, iron deficiency control (ID), daily low iron diet supplement (LDs), daily high iron diet supplement (HDs), weekly low iron supplement (LWs), and weekly high iron supplement (HWs). RESULTS: After 12 weeks, bone marrow stainable iron was seldom in ID group, and ample in supplement groups. The proportions of iron staining of bone marrow smear in supplement groups were more than 30%. Bone marrow cells in all rats were hyperplastic or active hyperplastic. CONCLUSIONS: Daily high iron supplement or once weekly high iron supplement were safe to bone marrow hemopoiesis in rats.
