{"title":"艾氯胺酮抑制心肌细胞自噬,通过下调瞬时受体电位香兰素1表达,抑制Ca2+/CaMKKβ/AMPK/mTOR通路,减轻心肌细胞缺氧/再氧化损伤。","authors":"Y Zhang, Q M Lu, H C Hu, C C Yang, Q H Zhao","doi":"10.26402/jpp.2025.2.05","DOIUrl":null,"url":null,"abstract":"<p><p>This research sought to determine the influence of esketamine (ESK) on hypoxia/reoxygenation (H/R) injury in cardiomyocytes by blocking autophagy via the transient receptor potential vanilloid type 1 (TRPV1)/ Ca<sup>2+</sup>/ calmodulin-dependent protein kinase β (CaMKKβ)/ adenosine monophosphate (AMP)-activated protein kinase (AMPK)/ mammalian target of rapamycin (mTOR) pathway. H9c2 cardiomyocytes were hypoxic for 4 h and reoxygenated for 6 h. H9c2 cells were pretreated with ESK (30 μg/ml) before hypoxia. H9c2 cells were transfected with plasmid vectors that interfered with TRPV1 or CaMKKβ, and the success of the transfections was verified by RT-qPCR. Cell viability was detected by MTT assay; apoptosis was detected by flow cytometry; intracellular Ca<sup>2+</sup> concentration ([Ca<sup>2+</sup>]i) was assessed using fluorescent dye Fluo-3 AM/Pluronic F127, and LC3-I, LC3-II, Beclin-1, and CaMKKβ/AMPK/mTOR-related proteins were detected by Western blot. In results: ESK treatment inhibited H/R-induced cell injury, cellular autophagy, and [Ca<sup>2+</sup>]i elevation. Induction of autophagy or [Ca<sup>2+</sup>]i elevation attenuated the ameliorative effect of ESK on H/R-induced cell injury. Upregulating TRPV1 attenuated We conclude that ESK-induced protection against H/R injury, as well as reduced the effect on the CaMKKβ/AMPK/mTOR pathway. ESK attenuates H/R cardiomyocyte injury by hindering autophagy through the TRPV1/Ca<sup>2+</sup>/CaMKKβ/AMPK/mTOR pathway.</p>","PeriodicalId":50089,"journal":{"name":"Journal of Physiology and Pharmacology","volume":"76 2","pages":""},"PeriodicalIF":2.0000,"publicationDate":"2025-04-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"0","resultStr":"{\"title\":\"Inhibition of autophagy by esketamine attenuates hypoxia/reoxygenation injury in cardiomyocytes via inhibition of Ca<sup>2+</sup>/CaMKKβ/AMPK/mTOR pathway by down-regulation of transient receptor potential vanilloid 1 expression.\",\"authors\":\"Y Zhang, Q M Lu, H C Hu, C C Yang, Q H Zhao\",\"doi\":\"10.26402/jpp.2025.2.05\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"<p><p>This research sought to determine the influence of esketamine (ESK) on hypoxia/reoxygenation (H/R) injury in cardiomyocytes by blocking autophagy via the transient receptor potential vanilloid type 1 (TRPV1)/ Ca<sup>2+</sup>/ calmodulin-dependent protein kinase β (CaMKKβ)/ adenosine monophosphate (AMP)-activated protein kinase (AMPK)/ mammalian target of rapamycin (mTOR) pathway. H9c2 cardiomyocytes were hypoxic for 4 h and reoxygenated for 6 h. H9c2 cells were pretreated with ESK (30 μg/ml) before hypoxia. H9c2 cells were transfected with plasmid vectors that interfered with TRPV1 or CaMKKβ, and the success of the transfections was verified by RT-qPCR. Cell viability was detected by MTT assay; apoptosis was detected by flow cytometry; intracellular Ca<sup>2+</sup> concentration ([Ca<sup>2+</sup>]i) was assessed using fluorescent dye Fluo-3 AM/Pluronic F127, and LC3-I, LC3-II, Beclin-1, and CaMKKβ/AMPK/mTOR-related proteins were detected by Western blot. In results: ESK treatment inhibited H/R-induced cell injury, cellular autophagy, and [Ca<sup>2+</sup>]i elevation. Induction of autophagy or [Ca<sup>2+</sup>]i elevation attenuated the ameliorative effect of ESK on H/R-induced cell injury. Upregulating TRPV1 attenuated We conclude that ESK-induced protection against H/R injury, as well as reduced the effect on the CaMKKβ/AMPK/mTOR pathway. ESK attenuates H/R cardiomyocyte injury by hindering autophagy through the TRPV1/Ca<sup>2+</sup>/CaMKKβ/AMPK/mTOR pathway.</p>\",\"PeriodicalId\":50089,\"journal\":{\"name\":\"Journal of Physiology and Pharmacology\",\"volume\":\"76 2\",\"pages\":\"\"},\"PeriodicalIF\":2.0000,\"publicationDate\":\"2025-04-01\",\"publicationTypes\":\"Journal Article\",\"fieldsOfStudy\":null,\"isOpenAccess\":false,\"openAccessPdf\":\"\",\"citationCount\":\"0\",\"resultStr\":null,\"platform\":\"Semanticscholar\",\"paperid\":null,\"PeriodicalName\":\"Journal of Physiology and Pharmacology\",\"FirstCategoryId\":\"3\",\"ListUrlMain\":\"https://doi.org/10.26402/jpp.2025.2.05\",\"RegionNum\":4,\"RegionCategory\":\"医学\",\"ArticlePicture\":[],\"TitleCN\":null,\"AbstractTextCN\":null,\"PMCID\":null,\"EPubDate\":\"2025/5/5 0:00:00\",\"PubModel\":\"Epub\",\"JCR\":\"Q3\",\"JCRName\":\"PHYSIOLOGY\",\"Score\":null,\"Total\":0}","platform":"Semanticscholar","paperid":null,"PeriodicalName":"Journal of Physiology and Pharmacology","FirstCategoryId":"3","ListUrlMain":"https://doi.org/10.26402/jpp.2025.2.05","RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"2025/5/5 0:00:00","PubModel":"Epub","JCR":"Q3","JCRName":"PHYSIOLOGY","Score":null,"Total":0}
Inhibition of autophagy by esketamine attenuates hypoxia/reoxygenation injury in cardiomyocytes via inhibition of Ca2+/CaMKKβ/AMPK/mTOR pathway by down-regulation of transient receptor potential vanilloid 1 expression.
This research sought to determine the influence of esketamine (ESK) on hypoxia/reoxygenation (H/R) injury in cardiomyocytes by blocking autophagy via the transient receptor potential vanilloid type 1 (TRPV1)/ Ca2+/ calmodulin-dependent protein kinase β (CaMKKβ)/ adenosine monophosphate (AMP)-activated protein kinase (AMPK)/ mammalian target of rapamycin (mTOR) pathway. H9c2 cardiomyocytes were hypoxic for 4 h and reoxygenated for 6 h. H9c2 cells were pretreated with ESK (30 μg/ml) before hypoxia. H9c2 cells were transfected with plasmid vectors that interfered with TRPV1 or CaMKKβ, and the success of the transfections was verified by RT-qPCR. Cell viability was detected by MTT assay; apoptosis was detected by flow cytometry; intracellular Ca2+ concentration ([Ca2+]i) was assessed using fluorescent dye Fluo-3 AM/Pluronic F127, and LC3-I, LC3-II, Beclin-1, and CaMKKβ/AMPK/mTOR-related proteins were detected by Western blot. In results: ESK treatment inhibited H/R-induced cell injury, cellular autophagy, and [Ca2+]i elevation. Induction of autophagy or [Ca2+]i elevation attenuated the ameliorative effect of ESK on H/R-induced cell injury. Upregulating TRPV1 attenuated We conclude that ESK-induced protection against H/R injury, as well as reduced the effect on the CaMKKβ/AMPK/mTOR pathway. ESK attenuates H/R cardiomyocyte injury by hindering autophagy through the TRPV1/Ca2+/CaMKKβ/AMPK/mTOR pathway.
期刊介绍:
Journal of Physiology and Pharmacology publishes papers which fall within the range of basic and applied physiology, pathophysiology and pharmacology. The papers should illustrate new physiological or pharmacological mechanisms at the level of the cell membrane, single cells, tissues or organs. Clinical studies, that are of fundamental importance and have a direct bearing on the pathophysiology will also be considered. Letters related to articles published in The Journal with topics of general professional interest are welcome.