Difference between revisions of "Team:BIT-China/H2O2DecompositionModel"

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                     some natural reduction molecular (like Glutathione) directly react with H<sub>2</sub>O<sub>2</sub>.
 
                     some natural reduction molecular (like Glutathione) directly react with H<sub>2</sub>O<sub>2</sub>.
 
                     And slow degradation
 
                     And slow degradation
                     is led by enzymatic degradation.(一篇参考文献)
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                     is led by enzymatic degradation.<sup onmouseover="tooltip.pop(this, '#tip-1', {position:1, offsetX:-20, effect:'slide',hideDelay: 10})"
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                    style="color:#38679a;">[1]</sup>
 
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                         1) The membrane permeability (k1).(三篇参考文献)
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                         1) The membrane permeability (k1).<sup onmouseover="tooltip.pop(this, '#tip-2', {position:1, offsetX:-20, effect:'slide',hideDelay: 10})"
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                        style="color:#38679a;">[2]</sup><sup onmouseover="tooltip.pop(this, '#tip-3', {position:1, offsetX:-20, effect:'slide',hideDelay: 10})"
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                        style="color:#38679a;">[3]</sup><sup onmouseover="tooltip.pop(this, '#tip-4', {position:1, offsetX:-20, effect:'slide',hideDelay: 10})"
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                        style="color:#38679a;">[4]</sup>
 
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                     </li>
 
                     <li>
 
                     <li>
                         2) Intracellular slow degradation rate (k2).(一篇参考文献)
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                         2) Intracellular slow degradation rate (k2).<sup onmouseover="tooltip.pop(this, '#tip-5', {position:1, offsetX:-20, effect:'slide',hideDelay: 10})"
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                        style="color:#38679a;">[5]</sup>
 
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                     </li>
 
                     <li>
 
                     <li>
 
                         3) Intracellular fast degradation rate (k3) dependent upon a finite capacity (g), e.g. NADPH
 
                         3) Intracellular fast degradation rate (k3) dependent upon a finite capacity (g), e.g. NADPH
                         levels in naive cells.(一篇参考文献)
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                         levels in naive cells.<sup onmouseover="tooltip.pop(this, '#tip-6', {position:1, offsetX:-20, effect:'slide',hideDelay: 10})"
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                        style="color:#38679a;">[6]</sup>
 
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                <a>Altıntaş A, Davidsen K, Garde C, et al. High-resolution kinetics and modeling of hydrogen peroxide degradation in live cells.[J]. Free Radical Biology & Medicine, 2016, 101:143-153.
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                <a>Branco M R, Marinho H S, Cyrne L, et al. Decrease of H<sub>2</sub>O<sub>2</sub> plasma membrane permeability during adaptation to H<sub>2</sub>O<sub>2</sub> in Saccharomyces cerevisiae.[J] . Journal of Biological Chemistry, 2004, 279(8):6501
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                <a>Henzler T, Steudle E. Transport and metabolic degradation of hydrogen peroxide in Chara corallina: model calculations and measurements with the pressure probe suggest transport of H(2)O(2) across water channels.[J]. Journal of Experimental Botany, 2000, 51(353):2053-2066.
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                <a>Bienert G P, Schjoerring J K, Jahn T P. Membrane transport of hydrogen peroxide[J]. Biochimica Et Biophysica Acta, 2006, 1758(8):994-1003.
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                <a>Branco M R, Marinho H S, Cyrne L, et al. Decrease of H<sub>2</sub>O<sub>2</sub> plasma membrane permeability during adaptation to H<sub>2</sub>O<sub>2</sub> in Saccharomyces cerevisiae.[J]. Journal of Biological Chemistry, 2004, 279(8):6501.
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                <a>Altıntaş A, Davidsen K, Garde C, et al. High-resolution kinetics and modeling of hydrogen peroxide degradation in live cells.[J]. Free Radical Biology & Medicine, 2016, 101:143-153.
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Revision as of 11:08, 17 October 2018

In our experiment, we found it hard to measure the intracellular H2O2 concentration, so we controlled the external H2O2 concentration in culture to confirm our system' function. Although the H2O2 can entry the cell quickly through simple diffusion, we needed to build up a model to simulate the intracellular H2O2. At the same time, this model will provide a base for roGFP2-Orp1 Micheali equation model.

Firstly, in order to describe the process in cells more accurately, three reactions in this process are considered: diffusion, fast degradation and slow degradation. The fast degradation is caused by some natural reduction molecular (like Glutathione) directly react with H2O2. And slow degradation is led by enzymatic degradation.[1]

Fig.1 The kinetic model for H2O2 degradation

$$\frac{\mathrm{d_{\mathit{p}_{e}}} }{\mathrm{d} t}=k_{1}(p_{i}-p_{e})v$$

$$\frac{\mathrm{d_{\mathit{p}_{i}}} }{\mathrm{d}t}=k_{1}(p_{e}-p_{i})-(k_{2}+k_{3}\frac{g}{k_{g}+g})p_{i}$$

$$\frac{\mathrm{d_{\mathit{g}}} }{\mathrm{d} t}=-k_{3}(\frac{g}{k_{g}+g})p_{i}$$

The ODE model accounted for intra- and extracellular H2O2 levels, and the consumption of a finite antioxidant capacity using three rates:

  • 1) The membrane permeability (k1).[2][3][4]
  • 2) Intracellular slow degradation rate (k2).[5]
  • 3) Intracellular fast degradation rate (k3) dependent upon a finite capacity (g), e.g. NADPH levels in naive cells.[6]

We reviewed the above literature and combined the results of the wet experiment, as shown in Fig.3, to determine the parameters required for the model simulation, listed in the table below.

Table 1. Description of Variables Used in The Kinetic Model.

Variable Explanation Value
k1 Membrane Permeability 11
k2 Slow Degradation Rate 8
k3 Fast Degradation Rate 55
G Finite Capacity Substance 60
pi Intracellular/Cytosolic [H2O2] /
pe Extracellular [H2O2] /
V Ratio of Cell Volume to Media 0.005

We used MATLAB to simulate the kinetics of extracellular and intracellular H2O2 changes over time. (Fig. 2)

Fig.2 Simulation result

To verify the feasibility of the model and to confirm this dynamic process, we did some experiments using our system's output, roGFP2-Orp1. The results show that the fluorescence ratio value I405/I488, an oxidant condition indicator, decreased by time when S. cerevisiae exposed to different concentrations of H2O2. This ratio's decreasing stands for the increasing of intercellular H2O2 concentration. It also stopped at a stable concentration at last, which's correspond to our model's result.

Fig.3 Fluorescence ratio value changes with time

In this model, we successfully simulated the kinetics of extracellular and intracellular H2O2 changes over time. Furthermore, the dynamic process of fluorescence ratio value by time was detected by using roGFP2-Orp1 green fluorescent protein in wet experiment, and the feasibility of the model was further verified.