Melatonin inhibits embryonic rat H9c2 cells growth through induction of apoptosis and cell cycle arrest via PI3K‐AKT signaling pathway

Abstract

Background Our recent epidemiological study revealed that maternal sleep during the periconceptional period should be involved in the risk of congenital heart disease (CHD) in offspring. Melatonin, a sleep related hormone, has been suggested to play a crucial role in embryonic development based on the emerging evidence. In this study, we set out to assess the effect of melatonin on the embryonic cardiac cell growth and to explore the underlying mechanisms. Methods We observed the effect of different gradient doses of melatonin as 10, 100, or 1,000 μM on cell proliferation in H9c2 embryonic rat cardiac cells. Furthermore, flow cytometry was applied to evaluate the impact on apoptosis and cell cycle. RNA‐seq was conducted to screen the changes in expression of mRNA and signaling pathways. Quantitative Real‐Time‐PCR (qRT‐PCR) was then conducted to validate the results. Results It was observed that melatonin could inhibit H9c2 cell growth, at the doses of 100 and 1,000 μM, but not at 10 μM. Moreover, melatonin ranged from 100 to 1,000 μM could instigate cell cycle arrest at G1 phase and simulate apoptosis, in a dose‐dependent manner. In addition, melatonin was found to down‐regulate the expression of a number of genes, which are related to heart development (SPARC, IFITM3, TNNT2, LOX), and PI3K‐Akt signaling pathway activation (FN1, HSP90B1, THBS1, MFGE8, and CLU). Conclusions Our findings suggested that high level of melatonin could be capable of inhibiting growth through the induction of apoptosis and cell cycle arrest via PI3K‐AKT signaling pathway, thereby interfering with embryonic heart development. Considering this study is based on H9c2 embryonic rat cardiac cells, future additional studies using human embryonic cardiac cell are warranted.

Genetic Diagnostic Yield and Novel Causal Genes of Congenital Heart Disease

“…The “protein digestion and absorption” pathway was also enriched in our results with FDR = 0.0316. In a study by Yu et al (2022) proved that gene PDTLN1 caused cardiac developmental defects in zebrafish via suppressing the PI3K/AKT signaling pathway, and another study by Zhao et al (2021) discovered that a high level of melatonin could inhibit growth by inducing apoptosis and cell cycle arrest via PI3K-AKT signaling pathway, thereby interfering with embryonic heart development. In a review by Liu et al (2019) summarized recent research progresses of actin cytoskeleton in the deployment process of mouse second heart field (SHF) progenitor cells and revealed that actin cytoskeleton played a significant role in mouse SHF development.…”

Bioinformatics analysis and validation of differentially expressed miRNA in the sperm of partners of patients with unexplained recurrent miscarriages

“…Ding et al [18] discovered that circCREBBP targets miR-143-3p and promotes boar sperm survival and vitality through the PI3K-Akt signalling pathway. Zhao et al [19] revealed that high melatonin levels may induce cell apoptosis, arrest the cell cycle, and inhibit cell growth through the PI3K-AKT signalling pathway, thus interfering with rat embryonic heart development. Nucleotide-binding oligomerisation domains 1 and 2 (NOD1 and NOD2) in the villous tissue of patients with RSMs can activate the MAPK/p38 pathway, thereby inhibiting the invasion of trophoblast cells and leading to RSM occurrence [20].…”

Identification and external validation of the hub genes associated with cardiorenal syndrome through time-series and network analyses

“…The vital roles of FN1 and POSTN in heart and kidney disease have previously been reported. Wang et al found that FN1 was associated with immune infiltration in diabetic nephropathy [ 43 ], Su et al reported that FN1 could regulate the process of renal fibrosis [ 44 ], Zhao et al disclosed that FN1 was involved in rat H9C2 cardiomyocyte growth by regulating cell cycle arrest [ 45 ], and Patel et al discovered that the expression of FN1 increased in patients with sudden cardio death [ 46 ]. Cardiofibrosis has a significant influence on the prognosis of heart diseases, which can be regulated by POSTN [ 47 , 48 ].…”