The Study Of Protection Mechanism Of Sweetener Erythritol Against Oxidative Damage Induced By H₂O₂ In PC12 Cells

[Objective]Research whether sweetener erythritol can into PC 12 cells, whether erythritol can protected PC 12 cells that oxidative damage induced by H2O2, and whether erythritol affected e xpression of Caspase-9, Caspase-12, Hsp70, TXNIP in the Mitochondrial stress and endoplasm ic reticulum stress.The experiment can provide a reference for clinical research.[Methods] 1. PC 12 cells was cultured in RPMI-1640 medium (Contain 10% fetal bovine serum,100 U/mL penicillin streptomycin), Extract the logarithmic phase of PC 12 cells, to deal with PC 12 cells in different concentration of erythritol, preparing the intracellular and extracellular fluid of PC 12 cells, determining the erythritol content of the intracellular and extracellular fluid of PC 12 cells by RP-HPLC(refractive index detector).2. The model of PC 12 cells of oxidative Damage induced by H2O2, observing the cells morphology, determining the cells survival rate (determined by MTT method), the LDH leakage and GSH content, and researching the protection effects of Erythritol against Oxidative Damage induced by H2O2 in PC 12 cells.3. Determining the effects of expression of Caspase-9、Caspase-12、 Hsp70 and TXNIP with erythritol by Western-blot analysis.[Results] 1. We can detect the chromatographic peak of erythritol in the PC 12 cells, With the erythritol concentration increasing in the extracellular fluid, the erythritol content of intracellular fluid is gradually increased in the range of 0.4mg/mL to 2.0mg/mL concentration of erythritol 2. Erythritol can reduce H2O2 induce oxidative Damage in the PC12 cells, increase the cells survival rate (p<0.05), reduce the content of LDH leakage (p<0.01) and increase the content of GSH (p<0.01).3. H2O2 induce oxidative Damage in the PC 12 cells 2 hours later, the expression of Caspase-9 increase in model group, the high dose content of erythritol has the reduction trend of expression of Caspase-9 (p<0.05) the expression of Caspase-12 obviously increase in model group (p<0.05), the high dose content of erythritol group has the reduction trend of expression of Caspase-12 but it has no statistical significance, there is no obvious effects for model groups on the expression of Hsp70 and TXNIP what compare with normal groups.[Conclusion] 1. The content changes of erythritol in intracellular and extracellular fluid of PC 12 cells revealed that erythritol can be into PC 12 cells, the content of erythritol in intracellular fluid increased with the content of erythritol in extracellular fluid increased 2. the erythritol can increase the cells survival rate of PC 12 cells with reduce the content of LDH leakage and increase the content of GSH and erythritol can reduce expression of Caspase-9、 Caspase-12, and against H2O2 induce oxidative Damage in the PC 12 cells 3. The erythritol of sweetener can into the PC 12 cells, and erythritol have protection effects for PC 12 cells with H2O2 induce oxidative Damage by mitochondrial pathway et al.