Effects of Circulating HMGB-1 and Histones on Cardiomyocytes–Hemadsorption of These DAMPs as Therapeutic Strategy after Multiple Trauma

Figures

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  Figure 1. Decrease in cardiomyocytes viability, apoptosis and alterations of calcium handling in the presence of HMGB-1 and histones: Cell viability of human cardiomyocytes (Luminescence in counts/sec) in the presence of 20 µg/mL histones (His) and 100 ng/mL HMGB-1 (37 °C, 6 h) compared to the control (PBS) (A). Caspase-3/7 activity (counts/sec) in human CMs in the presence of histones and HMGB-1 (B). Mean Height calcium signals (A.U) in the presence of histones and HMGB-1 compared to the control (C). Mean rise time (sec) (D) and mean decay time (sec) (E) of calcium peaks of human CMs in the presence of histones and HMGB-1. Frequency of calcium signals (sec) in human cardiomyocytes treated 20 µg/mL histones and 100 ng/mL HMGB-1 (F). Results are presented as mean ± SEM, for all experiments n = 6. Indicated results were significant * p < 0.05.

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  Figure 2. Decrease in HL-1 cell viability and metabolic activity in the presence of different HMGB-1 concentrations. HL-1 cell viability in the presence of different HMGB-1 concentrations after 6 h of incubation compared to control-HL-1 cell viability (A). Metabolic activity measured in HL-1 cells in the presence of different HMGB-1 concentrations incubated for 24 h compared to controls (B). n = 6. Results were significant * p < 0.05.

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  Figure 3. Metabolic alterations of human cardiomyocytes (CM) in the presence of histones: Basal respiration (OCR in pmol/m in/Norm. unit) of human CMs in the presence of 20 µg/mL histones (37 °C, 6 h) (A). Maximal respiratory capacity (OCR in pmol/m in/Norm. unit) of human CMs in the presence of 20 µg/mL histones compared to the control (PBS) (B). Spare respiration (OCR in pmol/m in/Norm. unit) of human CMs in the presence of 20 µg/mL histones (37 °C, 6 h) (C). Basal respiration (D), max respiratory capacity (E) and spare respiration (F) of human CM in the presence of 100 ng/mL HMGB-1. Results are presented as mean SEM, for all experiments n = 6. Results are significant * p < 0.05, statistical analysis with t-test.

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  Figure 4. Hemadsorption—a therapeutic option to eliminate systemic extracellular histones. (A). Hemadsoption of different concentrations of histones (700, 500, 300, 200, 100, 50, 25 µg/mL) presented as percentage of reduction (%) after 6 hrs incubation time; for all concentrations n = 6. (B). Histone levels in blood plasma of 22 multiple injured humans (µg/mL) in the Emergency room (pre) and after hemadsorption for 6 h (post). Results were presented as SEM, * p < 0.05.