艾叶挥发油诱导白念珠菌凋亡(1)
[摘要]研究艾叶挥发油(Artemisia argyi essential oil，AAEO)誘导白念珠菌凋亡的活性。实验采用微量液基稀释法检测AAEO对白念珠菌(Candida albicans)SC5314的MIC；采用XTT还原法检测AAEO对白念珠菌SC5314代谢的影响；采用流式细胞仪检测AAEO对白念珠菌SC5314细胞内活性氧(reactive oxygen species，ROS)和线粒体膜电位(mitochondrial membrane potential，MMP)水平的影响；采用AnnexinV/PI法测定AAEO对白念珠菌SC5314细胞凋亡率的影响；采用荧光显微镜观察AAEO对白念珠菌SC5314细胞Caspase酶活性的影响；采用DAPI染色法观察AAEO对白念珠菌SC5314细胞核固缩的影响。结果显示，AAEO对白念珠菌SC5314的MIC为05 mL·L-1；XTT代谢活性显示AAEO对白念珠菌SC5314代谢呈剂量依赖性的抑制；>05 mL·L-1的AAEO干预后，白念珠菌SC5314细胞内活性氧水平显著升高、线粒体膜电位显著降低、凋亡比例明显增加、metacaspase活性显著升高、细胞核固缩。AAEO具有诱导白念珠菌细胞凋亡的活性，该作用可能与ROS积累和线粒体损伤相关。
, http://www.100md.com
[关键词]艾叶挥发油； 白念珠菌； 凋亡； 活性氧； 线粒体
Activity of essential oil extracted from Artemisia argyi in inducing
apoptosis of Candida albicans
SHI Gaoxiang1，2， WANG Tianming1，2， WU Shengbing4， WANG Yunxia1，2，SHAO Jing1，2， ZHOU Meiqi3*， WANG Changzhong1，2*
(1. School of Integrated Traditional Chinese and Western Medicine， Anhui University of Chinese Medicine， Hefei 230038， China；
, 百拇医药
2. Institute of Integrated Traditional and Western Medicine， Anhui Academy of Chinese Medicine， Hefei 230038， China；
3. Division of Science and Technology， Anhui University of Chinese Medicine， Hefei 230038， China；
4. Experimental Center of Anhui University of Chinese Medicine， Hefei 230038， China)
[Abstract]To explore the activity of essential oil extracted from Artemisia argyi (AAEO) in inducing the apoptosis of Candida albicans SC5314. The effect of AAEO on reactive oxygen species(ROS) and mitochondria membrane potential(MMP) of C. albicans SC5314 was detected by flow cytometry. Phosphatidylserine externalization was observed under fluorescence microscopic with AnnexinV/PI staining at the early stage of apoptosis in C. albicans. Metacaspase activity was observed under fluorescence microscopic with FITCVADFMK staining at the early stage of apoptosis in C. albicans. C. albicans morphology was observed by DAPI nuclear staining and fluorescence microscopy. After intervention with 0.5 mL·L-1 AAEO， apoptosis of C. albicans significantly increased， metacaspase activity increased， nuclear pyknosis and fragmentation， and intracellular ROS were significantly increased， and mitochondrial membrane potential decreased significantly. The certain concentrations of AAEO could induce the apoptosis of C. albicans.
, http://www.100md.com
[Key words]Artemisia argyi essential oil； Candida albicans； apoptosis； ROS； mitochondrial
临床上，皮肤发生病毒、细菌或真菌等病原体感染时，除了常规的全身或局部用药外，艾灸往往也能产生显著疗效[12]。艾灸具有药物作用、温热作用与近红外辐射作用三大作用机制，其中药物作用与其含有的挥发油密切相关[3]。课题组前期研究表明，艾叶挥发油(Artemisia argyi essential oil，AAEO)对临床常见皮肤致病性真菌尤其是白念珠菌具有较强的抗菌活性[4]，但其作用机制不详。有研究表明，植物挥发油可以通过诱导真菌凋亡而发挥其抗菌作用[5]，本实验着重探讨艾叶挥发油是否能通过凋亡方式发挥抗白念珠菌活性。, 百拇医药(施高翔汪天明吴生兵汪云霞邵菁周美启汪长中)
, http://www.100md.com
[关键词]艾叶挥发油； 白念珠菌； 凋亡； 活性氧； 线粒体
Activity of essential oil extracted from Artemisia argyi in inducing
apoptosis of Candida albicans
SHI Gaoxiang1，2， WANG Tianming1，2， WU Shengbing4， WANG Yunxia1，2，SHAO Jing1，2， ZHOU Meiqi3*， WANG Changzhong1，2*
(1. School of Integrated Traditional Chinese and Western Medicine， Anhui University of Chinese Medicine， Hefei 230038， China；
, 百拇医药
2. Institute of Integrated Traditional and Western Medicine， Anhui Academy of Chinese Medicine， Hefei 230038， China；
3. Division of Science and Technology， Anhui University of Chinese Medicine， Hefei 230038， China；
4. Experimental Center of Anhui University of Chinese Medicine， Hefei 230038， China)
[Abstract]To explore the activity of essential oil extracted from Artemisia argyi (AAEO) in inducing the apoptosis of Candida albicans SC5314. The effect of AAEO on reactive oxygen species(ROS) and mitochondria membrane potential(MMP) of C. albicans SC5314 was detected by flow cytometry. Phosphatidylserine externalization was observed under fluorescence microscopic with AnnexinV/PI staining at the early stage of apoptosis in C. albicans. Metacaspase activity was observed under fluorescence microscopic with FITCVADFMK staining at the early stage of apoptosis in C. albicans. C. albicans morphology was observed by DAPI nuclear staining and fluorescence microscopy. After intervention with 0.5 mL·L-1 AAEO， apoptosis of C. albicans significantly increased， metacaspase activity increased， nuclear pyknosis and fragmentation， and intracellular ROS were significantly increased， and mitochondrial membrane potential decreased significantly. The certain concentrations of AAEO could induce the apoptosis of C. albicans.
, http://www.100md.com
[Key words]Artemisia argyi essential oil； Candida albicans； apoptosis； ROS； mitochondrial
临床上，皮肤发生病毒、细菌或真菌等病原体感染时，除了常规的全身或局部用药外，艾灸往往也能产生显著疗效[12]。艾灸具有药物作用、温热作用与近红外辐射作用三大作用机制，其中药物作用与其含有的挥发油密切相关[3]。课题组前期研究表明，艾叶挥发油(Artemisia argyi essential oil，AAEO)对临床常见皮肤致病性真菌尤其是白念珠菌具有较强的抗菌活性[4]，但其作用机制不详。有研究表明，植物挥发油可以通过诱导真菌凋亡而发挥其抗菌作用[5]，本实验着重探讨艾叶挥发油是否能通过凋亡方式发挥抗白念珠菌活性。, 百拇医药(施高翔汪天明吴生兵汪云霞邵菁周美启汪长中)