艾烟对肺泡Ⅱ型上皮细胞A549线粒体膜电位及Bax/Bcl-2影响的研究
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 rea...
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| Published in: | 针灸推拿医学：英文版 Vol. 14; no. 5; pp. 305 - 310 |
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| Main Author: | |
| Format: | Journal Article |
| Language: | English |
| Published: |
2016
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| Subjects: | |
| Online Access: | Get full text |
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| Summary: | 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。 |
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| Bibliography: | 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）. Conclusion： Certain higher concentration of moxa smoke could reduce MTP and mRNA expression of the anti-apoptosis gene Bcl-2 in alveolar type Ⅱ epithelial A549 cells. Oxidative damage may be the important mechanism of apoptosis caused by the high concentration of moxa smoke solution, and further studies are necessary on the specific mechanisms. Moxibustion Therapy; Artemisia Argyi; Smoke; Smoke Inhalation Injury; Adverse Effects; Safety; Primary Cell Culture 31-1908/R Dou Chuan-zi , Wu Huan-gan , Ma Xiao-peng , Huang Yan , Zhao Ji-meng, Liu Hui-rong, Cui Yun-hua , Zhou Ci-li , Zhao Chen（ 1 Shanghai Research Institute of Acupuncture and Meridian, Shanghai 200030, China ;2 Shanghai University of Traditional Chinese Medicine, Shanghai 201203, China） |
| ISSN: | 1672-3597 1993-0399 |