伤害诱导白木香愈伤组织形成倍半萜过程中活性氧的产生和积累  

ROS Synthesis and Accumulation during Wound-induced Sesquiterpene Biosynthesis in Aquilaria sinensis Callus

作　　者：吕菲菲[1] 孙佩文 刘培卫 杨云[1] 肖梦君 徐艳红[1] 魏建和[1,2] LUFei-fei;SUN Pei-wen;LIU Pei-wei;YANG Yun;XIAO Meng-jun;XU Yan-hong;WEI Jian-he(Hainan Provincial Key Laboratory of Resources Conservation and Development of Southern Medicine&Key Laboratory of State Administration of Traditional Chinese Medicine for Agarwood Sustainable Utilization,Hainan Branch of the Institute of Medicinal Plant Development,Chinese Academy of Medical Sciences and Peking Union Medical College,Haikou 570311,China;Key Laboratory of Bioactive Substances and Resources Utilization of Chinese Herbal Medicine,Ministry of Education&National Engineering Laboratory for Breeding of Endangered Medicinal Materials,Institute of Medicinal Plant Development,Chinese Academy of Medical Sciences and Peking Union Medical College,Beijing 100193,China)

机构地区：[1]中国医学科学院北京协和医学院药用植物研究所海南分所,海南省南药资源保护与开发重点实验室,国家中医药管理局沉香可持续利用重点研究室,海口570311 [2]中国医学科学院北京协和医学院药用植物研究所,中草药物质基础与资源利用教育部重点实验室,濒危药材繁育国家工程实验室,北京100193

出　　处：《中国药学杂志》2019年第23期1939-1944,共6页Chinese Pharmaceutical Journal

基　　金：国家重点研发计划项目资助(2018YFC1706400);海南省自然科学基金项目资助(20163150);中组部“万人计划”项目资助(99950534);中国医学科学院医学与健康科技创新工程项目资助(2016-I2M-2-003)

摘　　要：目的探索伤害诱导白木香(Aquilaria sinensis)愈伤组织形成倍半萜过程中活性氧(reactive oxygen species,ROS)组分过氧化氢(hydrogen peroxide,H2O2),超氧阴离子(superoxide radical ion,O2^-),羟基自由基(hydroxyl radical,OH^-)对伤害刺激的响应强度。方法GG-MS分析白木香愈伤组织受伤害后倍半萜合成,荧光显微观察H2DCF染色后细胞内总ROS分布情况,生化反应检测细胞内H2O2、O2^-和OH^-含量变化。结果伤害处理愈伤组织细胞后,两种倍半萜合成并积累。ROS作为信号分子在白木香伤害诱导结香过程中爆发并积累,细胞内DCF荧光逐渐增强,3 d达到最高点后降低。H2O2、O2^-和OH^-含量显著增加,H2O2和O2^-对伤害响应早于OH^-,H2O2和O2^-在3 d时达到最大值后降低,OH^-在5 d后才达到最大值。结论ROS各组分作为信号分子均响应于伤害刺激,且各组分响应强度差异可协同调控白木香愈伤组织防御反应促进倍半萜合成。OBJECTIVE To explore the response intensity of reactive oxygen species(ROS)components including H2O2,O2^- and OH^-in the course of wound-induced sesquiterpenes formation in Aquilaria sinensis callus.METHODS The synthesis of sesquit-erpenes in wounded callus was analyzed by using GG-MS technique.Total ROS distribution in cells was observed by fluorescence microscope after H2DCF fluorescent probe staining.The H2O2,O2^- and OH^-contents were measured according to biochemical reaction principle.RESULTS Two sesquitterpenes were synthesized and accumulated in the wounded cells.As a signal molecule,ROS burst and accumulated in the process of wound-induced sesquiterpenes formation,DCF fluorescence was observed to increase gradually.DCF fluorescence reached peak at 3 d and then began to decrease.The response of H2O2 and O2^- to injury took place earlier than those of OH^-,H2O2 and O2^- reached the maximum value at 3 d and then decreased,while OH-reached the maximum value at 5 d.CON-CLUSION All ROS components respond to injury stimulation as signaling molecules,and the difference in response intensity of each component can synergistically regulate defense response to promote the synthesis of sesquiterpenes in A.sinensis callus.

关 键 词：白木香 沉香倍半萜 伤害 防御反应 活性氧 

分 类 号：R284[医药卫生—中药学]