【摘要】 目的:观察NaHS（H₂S供体）对ATP（三磷酸腺苷）诱导的PC12细胞存活率及膜通透性变化的影响,探讨H₂S对抗ATP致伤作用的可能机制。方法:高分化PC12细胞置于含有高浓度ATP的培养基中3 h制备细胞损伤模型。在ATP处理前30 min,把NaHS或CBX（生胃酮,Pannxin-1通道阻滞剂）加入培养基中,作为预处理。用CCK-8检测细胞存活率,Fura-2/AM荧光染料检测细胞[Ca²⁺]_i,YO-PRO-1或Lucifer Yellow（荧光黄）荧光染料检测RFU（胞内相对荧光单位）,研究NaHS干预后细胞存活率和膜通透性的变化。结果:ATP（3、5mmol/L）处理使PC12细胞存活显著降低,NaHS（200μmol/L）和CBX（10μmol/L）预处理均可显著拮抗ATP（3 mmol/L）诱导的细胞存活率下降。ATP（3、5 mmol/L）可使PC12细胞内[Ca²⁺]_i明显增加,而NaHS（200μmol/L）和CBX（10μmol/L）预处理可使ATP（3 mmol/L）诱导的胞内[Ca²⁺]_i的增幅明显降低。ATP（3、5mmol/L）使PC12细胞对YO-PRO-1和Lucifer Yellow通透明显增加,而NaHS（200μmol/L）和CBX（10μmol/L）预处理可显著阻遏ATP诱导的膜通透性增加。结论:Pannxin-1蛋白在ATP诱导PC12细胞膜孔形成中有重要作用,干预膜孔形成可能是NaHS拮抗ATP损伤作用的机制之一。更多还原

【Abstract】 Objective: To explore the possible mechanism of H₂S（ hydrogen sulfide） protection against adenosine triphosphate（ ATP） induced PC12 cell injury,the effect of NaHS（ a donor of H₂S） on survival or membrane permeability induced by ATP were observed. Methods: PC12 cells were treated with high concentration of ATP for 3 hours to set up the injured neuron-like cell model. NaHS or CBX（ carbenoxolone,Pannxin-1 receptor antagonist） were added into the medium for 30 minutes before the ATP treatment. The viability of PC12 cell was measured by CCK-8 assay. The[Ca²⁺]_i was detected by Fura-2/AM staining. Fluorescence changes（ RFU） following YO-PRO-1 or Lucifer Yellow uptake were monitored. So the effect of NaHS pretreatment on the cell survival rate and membrane permeability was studied. Results: ATP（ 3,5 mmol/L） made PC12 cell survival significantly reduced,but pretreatment with NaHS（ 200 μmol/L） or CBX（ 10 μmol/L） for 30 min could significantly antagonized cell survival rate drop induced by ATP（ 3 mmol/L）. ATP（ 3,5 mmol/L） made [Ca²⁺]_i in PC12 cell increased significantly,however,pretreatment with NaHS（ 200 μmol/L） or CBX（ 10 μmol/L） for 30 min could obviously depressed the [Ca²⁺]_i amplification induced by ATP（ 3 mmol/L）. ATP（ 3,5 mmol/L） made YO-PRO-1 and Lucifer Yellow transparent increased significantly in PC12 cell,but pretreatment with NaHS（ 200 μmol/L） or CBX（ 10 μmol/L） for 30 min could inhibited membrane permeability increase induced by ATP obviously. Conclusion: Pannexin-1 protein plays an important role in PC12 cell membrane pore formation induced by ATP at high concentration and the intervention of membrane pore formation may be one of the mechanisms of NaHS against ATP-induced injury.更多还原