| Objective:To study the protective effect of H2S on cerebral ischemia-reperfusion injury and its relationship with cell autophagy.Methods:To stimulate in vivo cerebral ischemia-reperfusion injury withSH-SY5Y cells culture in vitro and in rat models by MACO by dividinginto six groups: Control group, HR group, HR+10μM H2S group,HR+30μM H2S group, HR+60μM H2S group, HR+30μM H2Sgroup+Ly294002(PI3K inhibitor) group.1. To observe morphological changes of SH-SY5Y cells throughoptical microscope;2. To measure OD value of SH-SY5Y cells and compare survivalrates by MTT analysis;3. To measure infarct sizes of cerebral tissue in rats by TTC staining;4. To detect Beclin1, LC3B and Akt expression by Westernblotting both in rats and SH-SY5Y cells.Results:1. Compared to Control group, SH-SY5Y cells numbers of HRgroup decreased markedly（P＜0.05）. H2S pretreatment showed asignificant increase of SH-SY5Y cells numbers, survival rate had a verystrong capacity to dose-dependently increase, especially at highestconcentration (60μM H2S). Compared to HR group, every H2Spretreatment groups had a significant statistic differences（P＜0.05）. Compared to HR+30μM H2S group, LY294002pretreatment groupshowed decrease of SH-SY5Y cells numbers and had a significantstatistic differences（P＜0.05）.2. Compared to Control group, there is an obvious infarct lesion ofbrain tissue in HR group rats（P＜0.05）. H2S pretreatment showed asignificant decrease size of infarct area（P＜0.05）, infarct rate had a verystrong capacity to dose-dependently decrease, especially at highestconcentration (60μM H2S). Compared to HR group, every H2Spretreatment groups had a significant statistic differences（P＜0.05）.Compared to HR+30μM H2S group, LY294002pretreatment groupshowed increase size of infarct area and had a significant statisticdifferences（P＜0.05）.3. Compared to Control group, Beclin1expression of HR groupincreased markedly both in SH-SY5Y cells and in rats（P＜0.05）. H2Spretreatment showed a very strong capacity to dose-dependentlysignificant decrease of Beclin1expression, especially at highestconcentration (60μM H2S). Compared to HR group, every H2Spretreatment groups had a significant statistic differences（P＜0.05）.Compared to HR+30μM H2S group, LY294002pretreatment groupshowed decrease of Beclin1expressionand had a significant statisticdifferences（P＜0.05）.4. Compared to Control group, LC3B expression of HR groupincreased markedly both in SH-SY5Y cells and in rats（P＜0.05）. H2Spretreatment showed a very strong capacity to dose-dependentlysignificant decrease of LC3B expression, especially at highestconcentration (60μM H2S). Compared to HR group, every H2Spretreatment groups had a significant statistic differences（P＜0.05）. Compared to HR+30μM H2S group, LY294002pretreatment groupshowed decrease of LC3B expressionand had a significant statisticdifferences（P＜0.05）.5. Compared to Control group, Akt protein expression of HR grouphad no significant difference both in SH-SY5Y cells and in rats（P>0.05）.H2S pretreatment showed a very strong capacity to dose-dependentlysignificant increase of Akt protein, especially at highest concentration(60μM H2S). Compared to HR group, every H2S pretreatment groups hada significant statistic differences（P＜0.05）. Compared to HR+30μM H2Sgroup, LY294002pretreatment group showed decrease of Akt proteinexpression and had a significant statistic differences（P＜0.05）.Conclusion:1. Cerebral ischemia-reperfusion injury can be dose-dependentlyrelieved by exogenous H2S.2. Cerebral ischemia-reperfusion injury can be prevented byexogenous H2S through inhibiting cell autophagy.3. Activation of PI3K/Akt signaling pathway may be involved inprotective mechanism of exogenous H2S on cerebral ischemia-reperfusion injury by inhibiting cell autophagy. |
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