Protective Effect of Metformin on Sepsis Myocarditis in Zebrafish

Figures

• 

  Figure 1.. Survival curve of zebrafish in different groups. A, Survival curves, n = 20 fish per group. The fish were soaked in different concentrations of Escherichia coli solution for 2 hours, 10 × 108 CFU/mL, 20 × 108 CFU/mL, 40 × 108 CFU/mL, 80 × 108 CFU/mL, and 100 × 108 CFU/mL, respectively. B, Survival curves, n = 20 fish per group. The fish were soaked in different concentrations of E coli solution for 3 hours, 10 × 108 CFU/mL, 20 × 108 CFU/mL, 40 × 108 CFU/mL, 80 × 108 CFU/mL, and 100 × 108 CFU/mL, respectively. C, Survival curves, n = 20 fish per group. The fish were soaked in different concentrations of E coli solution for 3 hours, 2 × 108 CFU/mL, 4 × 108 CFU/mL, 6 × 108 CFU/mL, 8 × 108 CFU/mL, and 10 × 108 CFU/mL, respectively. D, Survival curves, n = 20 fish per group. The fish were first soaked in different concentrations of metformin for 2 hours, second, soaked in 10 × 108 CFU/mL E coli for 3 hours. E, Survival curves, n = 20 fish per group. The fish were first soaked in different concentrations of metformin for 3 hours, second, soaked in 10 × 108 CFU/mL E coli for 3 hours. F, Survival curves, n = 20 fish per group. The fish were first soaked in different concentrations of…

• 

  Figure 2.. Effect of metformin on heart of zebrafish with sepsis myocarditis. A, Effect of metformin on swelling and congestion of zebrafish heart with sepsis myocarditis, the whole hearts were circled by red zones. The area of red zone represented as the severity of myocarditis. B, Effect of metformin on heart area of zebrafish with sepsis myocarditis, the area of red zones calculated by computer software were statistically analyzed (**P < .05; ***P < .01, n = 10), (Escherichia coli group vs PBS group, Met + E coli vs E coli group). C, Effect of metformin on heart rate of zebrafish with sepsis myocarditis, the heart rate of zebrafish recorded on video were statistically analyzed (**P < .05; ***P < .01, n = 10), (E coli group vs PBS group, Met + E coli vs E coli group).

• 

  Figure 3.. Effect of metformin on cardiac neutrophils in zebrafish with sepsis myocarditis. A, Cardiac neutrophils of different groups of zebrafish recorded by repeated experiments with Tg (mpx: EGFP) transgenic zebrafish, the neutrophils labeled by green fluorescent protein could be visually counted. And the cardiac neutrophils were amplificated and shown in the upper right corner of each group image. B, Statistical analysis of neutrophils in different groups of hearts (**P < .05; ***P < .01, n = 10; E coli group vs PBS group, Met + E coli vs E coli group).

• 

  Figure 4.. Expression of cytokines in different groups. A, Expression of TNF-α in different groups, qRT-PCR showed that metformin could significantly reduce the expression of TNF-α (**P < .05; ***P < .01; E coli group vs PBS group, Met + E coli vs E coli group). B, Expression of IL-6 in different groups, qRT-PCR showed that metformin could significantly reduce the expression of IL-6 (**P < .05; ***P < .01; E coli group vs PBS group, Met + E coli vs E coli group). C, Expression of TGF-β in different groups, qRT-PCR showed that metformin could significantly enhance the expression of TGF-β. (**P < .05; ***P < .01; E coli group vs PBS group, Met + E coli vs E coli group). D, Expression of IL-10 in different groups, qRT-PCR showed that metformin could significantly enhance the expression of IL-10 (**P < .05; ***P < .01; E coli group vs PBS group, Met + E coli vs E coli group). IL-6 indicates interleukin-6; IL-10, interleukin-10; PBS, phosphate-buffered saline; qRT-PCR, quantitative real-time reverse transcription polymerase chain reaction; TGF-β, transforming growth factor-β; TNF-α, tumor necrosis factor-α.