Identifying Involvement of H19-miR-675-3p-IGF1R and H19-miR-200a-PDCD4 in Treating Pulmonary Hypertension with Melatonin

Abstract

Non-coding RNAs play an important role in the pathogenesis of pulmonary arterial hypertension (PAH). The aim of this study was to characterize the therapeutic role of melatonin as well as the underlying molecular mechanism (its effects on the expression of H19 and its downstream signaling pathways) in the treatment of PAH. Real-time PCR and western blot analysis were performed to evaluate the expression of H19, miR-200a, miR-675, insulin-like growth factor-1 receptor (IGF1R), and programmed cell death 4 (PDCD4). The value of systolic pulmonary artery pressure (SPAP) and the ratio of medial thickening in the monocrotaline (MCT) group were increased, whereas the melatonin treatment could decrease these values to some extent. The weights of RV (right ventricle), LV (left ventricle) + IVS (interventricular septal), and RV/(LV + IVS) in the MCT group were much higher than those in the MCT + melatonin and control groups. In addition, the expression of H19, miR-675, IGF1R mRNA, and IGF1R protein in the MCT group was the highest, whereas their expression in the control group was the lowest. The expression of miR-200, PDCD4 mRNA, and PDCD4 protein in the MCT group was the lowest, whereas their expression in the control group was the highest. Furthermore, H19 directly suppressed the expression of miR-200a, whereas miR-675-3p and miR-200a directly inhibited the expression of IGF1R and PDCD4, respectively. Finally, melatonin treatment inhibited cell proliferation; upregulated the expression of H19, miR-675-3p, and PDCD4; and downregulated the expression of miR-200a and IGF1R. This study demonstrated the role of H19-miR-675-3p-IGF1R- and H19-miR-200a-PDCD4-signaling pathways in the melatonin treatment of PAH.

Microbiota Imbalance Contributes to COPD Deterioration by Enhancing IL-17a Production via miR-122 and miR-30a

“…The total RNA content in each plasma sample collected from the mice models was isolated by using a Trizol kit assay kit (Invitrogen, Carlsbad, CA, USA) following the standard protocol provided by the kit manufacturer. 35 , 36 In the next step, the isolated RNA was subjected to real-time PCR to determine the relative expression of miR-10a, miR-26a, miR-30a, miR-34c, miR-99b, miR-122a, miR-123-prec, miR-124a-prec b , miR-125a-prec b , miR-140 s b , miR-145-prec, miR-146-prec b , miR-191-prec, miR-192, miR-219-prec, miR-222-prec, miR-223-prec, miR-122, and IL-17a mRNA by the calculation provided by the 2 −ΔΔCt method. The qPCR data for miRNAs were normalized to RNU48, while the qPCR data for IL-17a mRNA was normalized to GAPDH.…”

Melatonin’s Impact on Antioxidative and Anti-Inflammatory Reprogramming in Homeostasis and Disease

“…Melatonin also diminishes bleomycin-induced lung fibrosis in mice by inhibiting transforming growth factor β 1 (TGF-β1) [ 200 , 201 ] and it is protective in radiation-induced lung injury by suppressing the NOD-like receptor protein 3 (NLRP3) inflammasome [ 70 , 202 ]. Through its antioxidant and anti-inflammatory effects, melatonin is beneficial in pulmonary hypertension [ 203 , 204 , 205 ], enhances the NO-independent acetylcholine’s vasodilatory effect, decreases the systolic pressure of the right ventricle, and prevents right ventricular hypertrophy [ 206 ]. By diminishing TGF-β1 activity, apoptosis, and endoplasmic reticulum stress via Sirtuin-1 upregulation in animal studies, melatonin could be protective in chronic obstructive pulmonary disease (COPD) [ 72 , 207 ].…”

LncRNA H19 is involved in myocardial ischemic preconditioning via increasing the stability of nucleolin protein

“…Meanwhile, H19 has been proven to be the primary transcript for microRNA‐675 (Dykes & Emanueli, 2017; Luo et al, 2019; Park, Mitra, Rahat, Kim, & Pfeifer, 2017). H19/miR‐675 were found to be associated with different types of diseases (Fan, Peng, Liang, Yang, & Zhang, 2019; Luo et al, 2019; Muller et al, 2019; Wang et al, 2018). Besides, H19 also acts as a competing endogenous RNA for other microRNAs (Dykes & Emanueli, 2017).…”