Journal: Toxicology Research

Article Title: Dexmedetomidine plays a protective role in sepsis-associated myocardial injury by repressing PRMT5-mediated ferroptosis

doi: 10.1093/toxres/tfaf010

Figure Lengend Snippet: Erastin remarkably reversed the cardioprotective effects of Dex in septic cardiomyocytes. Septic cardiomyocytes were constructed by LPS treatment (10 μg/mL, 24 h) and then co-treated with Dex (10 μM) and ferroptosis inducer (Erastin, 10 μM). A) Cell viability was detected by CCK8 assay. B) LDH level in cardiomyocytes was assessed using LDH release assay kit. (C–E) TNF-α, IL-6 and IL-1β levels were measured using ELISA. F) Flow cytometry was performed to detect cell apoptosis. The measurement data were presented as mean ± SD. All data was obtained from at least three replicate experiments. * P < 0.05, * * P < 0.01, * * * P < 0.001.

Article Snippet: The mouse primary cardiomyocytes were purchased from Procell (Hubei, China).

Techniques: Construct, CCK-8 Assay, Lactate Dehydrogenase Assay, Enzyme-linked Immunosorbent Assay, Flow Cytometry

Journal: Toxicology Research

Article Title: Dexmedetomidine plays a protective role in sepsis-associated myocardial injury by repressing PRMT5-mediated ferroptosis

doi: 10.1093/toxres/tfaf010

Figure Lengend Snippet: The inhibitory roles of Dex on ferroptosis in septic cardiomyocytes were reversed by Erastin. Mouse primary cardiomyocytes were stimulated by 10 μg/mL LPS for 24 h, and then treated with Dex (10 μM) alone or co-treated with Dex plus 10 μM Erastin. A–C) The oxidative stress indicators including MDA, SOD and GSH in cardiomyocytes were determined using the corresponding kits. D) ROS level was measured by the DCFH-DA probe. E) Fe 2+ level was measured by the kit. F) Western blot was employed to detect GPX4 and SLC7A11 protein levels. The measurement data were presented as mean ± SD. All data was obtained from at least three replicate experiments. * P < 0.05, * * P < 0.01, * * * P < 0.001.

Article Snippet: The mouse primary cardiomyocytes were purchased from Procell (Hubei, China).

Techniques: Western Blot

Journal: Toxicology Research

Article Title: Dexmedetomidine plays a protective role in sepsis-associated myocardial injury by repressing PRMT5-mediated ferroptosis

doi: 10.1093/toxres/tfaf010

Figure Lengend Snippet: PRMT5 was upregulated in LPS-treated cardiomyocytes and could be suppressed by Dex in a m 6 A-depended manner. Mouse primary cardiomyocytes were stimulated by 10 μg/mL LPS for 24 h, and then treated with Dex (10 μM). A and B) The mRNA and protein levels of PRMT5 in cardiomyocytes were detected using qRT-PCR and western blot. C) The global m 6 A modification in cardiomyocytes was determined using the m 6 A RNA methylation quantification kit. D) The m 6 A site modifications on PRMT5 mRNA were predicted using the SRAMP database. E) The m 6 A modification level of PRMT5 mRNA was examined by me-RIP assay. F) Western blot was adopted to determine METTL3, METTL14, WTAP, ALKBH5 and FTO protein levels. G and H) The interaction between METTL3/FTO and PRMT5 was analyzed by RIP assay. The measurement data were presented as mean ± SD. All data was obtained from at least three replicate experiments. * P < 0.05, * * P < 0.01, * * * P < 0.001.

Article Snippet: The mouse primary cardiomyocytes were purchased from Procell (Hubei, China).

Techniques: Quantitative RT-PCR, Western Blot, Modification, Methylation

Journal: Toxicology Research

Article Title: Dexmedetomidine plays a protective role in sepsis-associated myocardial injury by repressing PRMT5-mediated ferroptosis

doi: 10.1093/toxres/tfaf010

Figure Lengend Snippet: PRMT5 upregulation reversed the cardioprotective effects of Dex in septic cardiomyocytes. A) qRT-PCR was performed to detect PRMT5 mRNA level in cardiomyocytes after lv-NC or lv-PRMT5 transfection. PRMT5 overexpression was induced in septic cardiomyocytes combined with Dex treatment. B) CCK8 assay was conducted to examine cell viability. C) LDH level in cardiomyocytes was measured by the LDH release assay kit. D–F) ELISA was employed to assess TNF-α, IL-6 and IL-1β levels. G) Cell apoptosis was determined using flow cytometry. The measurement data were presented as mean ± SD. All data was obtained from at least three replicate experiments. * P < 0.05, * * P < 0.01, * * * P < 0.001.

Article Snippet: The mouse primary cardiomyocytes were purchased from Procell (Hubei, China).

Techniques: Quantitative RT-PCR, Transfection, Over Expression, CCK-8 Assay, Lactate Dehydrogenase Assay, Enzyme-linked Immunosorbent Assay, Flow Cytometry

Journal: Toxicology Research

Article Title: Dexmedetomidine plays a protective role in sepsis-associated myocardial injury by repressing PRMT5-mediated ferroptosis

doi: 10.1093/toxres/tfaf010

Figure Lengend Snippet: Overexpression of PRMT5 diminished the inhibitory roles of Dex on sepsis-induced cardiomyocyte ferroptosis. PRMT5 overexpression was induced in septic cardiomyocytes combined with Dex treatment. A–C) The oxidative stress indicators including MDA, SOD and GSH in cardiomyocytes were determined using the corresponding kits. D) ROS level was measured by the DCFH-DA probe. E) Fe 2+ level was assessed using the kit. F) GPX4 and SLC7A11 protein levels in cardiomyocytes were determined by western blot. The measurement data were presented as mean ± SD. All data was obtained from at least three replicate experiments. * P < 0.05, * * P < 0.01, * * * P < 0.001.

Article Snippet: The mouse primary cardiomyocytes were purchased from Procell (Hubei, China).

Techniques: Over Expression, Western Blot