目的:观察艾灸生成物对肺泡Ⅱ型上皮细胞A549线粒体膜电位(mitochondrial transmembrane potential,MTP)、Bax/Bcl-2 m RNA表达的影响,进一步研究艾灸生成物对A549细胞的氧化损伤作用。方法:采用气体采样滤盒采集艾灸生成的烟气颗粒物,用细胞培养液稀释后依次配制质量浓度为0.05 mg/mL、0.1 mg/mL、0.2 mg/mL、0.3 mg/mL、0.4 mg/mL的艾烟提取物(moxa smoke extract,MSE)溶液干预A549细胞。采用JC-1染色法检测细胞MTP,荧光定量聚合酶链反应(polymerase chain reaction,PCR)法检测细胞Bax/Bcl-2 mRNA的表达。结果:与正常对照组细胞相比,0.3 mg/mL和0.4 mg/mL MSE干预组细胞MTP显著降低(P〈0.01),0.05 mg/mL、0.1 mg/mL和0.2 mg/mL MSE干预组细胞MTP无显著变化(P〉0.05);与0.05 mg/mL MSE干预组细胞相比,0.1 mg/mL、0.2 mg/mL、0.3 mg/mL和0.4 mg/mL MSE干预组细胞MTP显著降低(均P〈0.05);与0.1 mg/mL MSE干预组细胞比较,0.4 mg/mL MSE干预组细胞MTP显著降低(P〈0.01)。各浓度MSE干预组细胞Bax mRNA的表达与正常对照组相比均无显著性差异;细胞内Bcl-2 mRNA表达随MSE干预浓度的增加而减少,其中0.4 mg/mL和0.3 mg/mL MSE干预组细胞与正常对照组细胞Bcl-2 mRNA的表达相比显著降低(P〈0.05),0.4 mg/mL MSE干预组细胞Bcl-2mRNA的表达与0.05 mg/mL MSE干预组细胞内Bcl-2 mRNA的表达相比显著降低(P〈0.05)。结论:艾烟升高到一定浓度时可以降低肺泡Ⅱ型上皮细胞A549细胞内MTP,减少凋亡抑制基因Bcl-2 mRNA的表达,氧化损伤可能是高浓度艾烟溶液引起细胞凋亡的重要机制,具体机制仍需进一步研究。
Objective: To investigate the influence of moxibustion products on mitochondrial transmembrane potential (MTP) and mRNA expression of Bax/Bcl-2 in alveolar type Ⅱ epithelial A549 cells, and to further explore influence of moxibustion products on the oxidative damage of A549 cells. Methods: Smoke and particles generated by moxibustion were collected using the filter box for gas sampling. The moxa smoke extract (MSE) was diluted sequentially to the final concentrations of 0.05 mg/mL, 0.2 mg/mL, 0.2 mg/mL, 0.3 mg/mL and 0.4 mg/mL using the cell culture medium, and A549 cells were then intervened by the above MSE solution. Cell MTP was detected by JC-1 staining. Fluorescence quantitative polymerase chain reaction (PCR) was used to detect Bax/Bcl-2 mRNA expression of A549 cells. Results: Compared with cells in the normal control group, MTP was significantly decreased in cells of 0.3 mg/mL and 0.4 mg/mL MSE intervention groups (P〈0.01); while MTP showed no significant changes in cells of 0.05 mg/mL, 0.1 mg/mL and 0.2 mg/mL MSE intervention groups (P〉0.05); compared with cells in 0.05 mg/mL MSE intervention group, MTP was decreased significantly in cells of 0.1 mg/mL, 0.2 mg/mL, 0.3 mg/mL and 0.4 mg/mL MSE intervention groups (P〈0.05); compared with cells in 0.1 mg/mL MSE intervention group, MTP was decreased significantly in cells of 0.4 mg/mL MSE intervention group (P〈0.01). Bax mRNA expression of cells in each concentration of MSE intervention group all showed no significant difference compared to that in the normal control group; Bcl-2 mRNA expression of cells was reduced with the increase of MSE intervention concentration. Wherein, Bcl-2 mRNA expressions of cells in 0.4 mg/mL and 0.3 mg/mL MSE intervention groups were significantly reduced compared with that of cells in the normal control group (P〈0.05); Bcl-2 mRNA expression of cells in 0.4 mg/mL MSE intervention group was significantly reduced compared to that in 0.05 mg/mL MSE intervention group (P〈0.05). C