The protective effect and mechanism of melatonin on myocardial ischemic reperfusion injury

doi:10.11958/20231005

Abstract

Abstract:

Objective To investigate the protective effect of melatonin on myocardial ischemia reperfusion injury(IRI) by regulating Akt/FOXO4/BIM signaling pathway and its mechanism. Methods Myocardial IRI model was established, and cells were divided into the control group (CTRL group), the ischemia reperfusion group (IRI group), the melatonin treatment group (IRI+MEL group) and the Akt inhibitor group (IRI+MEL+MK2206 group). MTT method was used to detect the cell survival rate. The level of released LDH was measured with microplate reader. Flow cytometry was used to detect cell apoptosis. PCR method was used to detect the inflammatory gene levels of interleukin 6 (IL-6), tumor necrosis factor α (TNF-α) and monocyte chemotactic protein-1 (MCP-1). Western blot assay was used to detect the protein expression changes of p-Akt, p-FOXO4 and BIM. Results Compared with the CTRL group, the cell survival rate decreased and LDH content increased, the early stage and total apoptosis rates were increased, expression levels of IL-6, TNF-α and MCP-1 genes were increased, p-Akt protein expression decreased, and p-FOXO4 and BIM protein expression levels increased in the IRI group (P<0.05). Compared with the IRI group, melatonin therapy increased the cell survival rate and p-Akt protein expression, decreased LDH content, cell apoptosis, IL-6, TNF-α, MCP-1 gene level and p-FOXO4 and BIM protein expression (P<0.05). Compared with the IRI+MEL group, MK2206 partially reversed the protective effect of melatonin on IRI cardiomyocytes (P<0.05). Conclusion Melatonin has protective effect on ischemia reperfusion injured myocardial cells, and its mechanism may be related to the regulation of Akt/FOXO4/BIM signaling pathway to reduce apoptosis and cell inflammation.

Key words: melatonin, reperfusion injury, signal transduction, inflammation, myocardial infarction

CLC Number:

R542.2+2

Figures/Tables 7

References 16

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基因名称	引物序列（5′→3′）	产物大小/bp
IL-6	上游：ACAACCACGGCCTTCCCTACTT	129
	下游：CACGATTTCCCAGAGAACATGTG	129
TNF-α	上游：GTAGCCCACGTCGTAGCAAA	214
	下游：TAGCAAATCGGCTGACGGTG	214
MCP-1	上游：ACCTGCTGCTACTCATTCACC	238
	下游：AAGACCTTAGGGCAGATGCAG	238
GAPDH	上游：TTCACCACCATGGAGAAGGC	237
	下游：GGCATGGACTGTGGTCATGA	237

基因名称	引物序列（5′→3′）	产物大小/bp
IL-6	上游：ACAACCACGGCCTTCCCTACTT	129
	下游：CACGATTTCCCAGAGAACATGTG	129
TNF-α	上游：GTAGCCCACGTCGTAGCAAA	214
	下游：TAGCAAATCGGCTGACGGTG	214
MCP-1	上游：ACCTGCTGCTACTCATTCACC	238
	下游：AAGACCTTAGGGCAGATGCAG	238
GAPDH	上游：TTCACCACCATGGAGAAGGC	237
	下游：GGCATGGACTGTGGTCATGA	237

组别	细胞存活率/%	LDH释放量/（U/mL）
CTRL组	100.00±9.77	0.24±0.05
IRI组	50.49±6.89^a	0.58±0.08^a
IRI+MEL组	77.39±4.60^b	0.35±0.06^b
IRI+MEL+MK2206组	56.25±7.88^c	0.59±0.07^c
F	53.762^＊	44.228^＊

组别	细胞存活率/%	LDH释放量/（U/mL）
CTRL组	100.00±9.77	0.24±0.05
IRI组	50.49±6.89^a	0.58±0.08^a
IRI+MEL组	77.39±4.60^b	0.35±0.06^b
IRI+MEL+MK2206组	56.25±7.88^c	0.59±0.07^c
F	53.762^＊	44.228^＊

组别	早期凋亡率	晚期凋亡率	总凋亡率
CTRL组	5.18±2.25	2.37±0.30	7.55±2.48
IRI组	31.50±2.38^a	4.04±2.87	35.54±4.99^a
IRI+MEL组	13.35±5.96^b	3.04±0.58	16.38±6.53^b
IRI+MEL+MK2206组	31.13±1.91^c	2.41±0.40	33.55±1.64^c
F	40.525^＊	2.363	34.462^＊