东革内酯通过InR-PI3K-mTOR-Bcl-2通路诱导斜纹夜蛾卵巢细胞凋亡  

Eurycomalactone induces apoptosis of ovarian cells through the InR-PI3K-mTOR-Bcl-2 pathway

作　　者：温雪梅 秦子昕 路伟[2] 黄建辉 欧阳嘉敏 万妍 邵雪花[1] WEN Xuemei;QIN Zixin;LU Wei;HUANG Jianhui;OUYANG Jiamin;WAN Yan;SHAO Xuehua(Institute of Pomology,Guangdong Academy of Agricultural Sciences,Key Laboratory of Biology and Utilization of Tropical Fruit Trees in South Asia,Ministry of Agriculture and Rural Affairs,Key Laboratory of Tropical and Subtropical Fruit Tree Research of Guangdong Province,Guangzhou 510640,China;College of Agriculture,Xinjiang Agricultural University,Engineering Research Centre of Cotton,Ministry of Education,Key Laboratory for Monitoring and Safety Control of Crop and Forest Pests of University of Xinjiang Uygur Autonomous Region,Urumqi 830052,China;Agricultural Service Center,Tanzhou Town,Zhongshan City,Guangdong Province,Zhongshan 528400,China)

机构地区：[1]广东省农业科学院果树研究所,农业农村部南亚热带果树生物学与遗传资源利用重点实验室,广东省热带亚热带果树研究重点实验室,广州510640 [2]新疆农业大学农学院,棉花教育部工程研究中心,自治区农林有害生物监测与安全防控重点实验室,乌鲁木齐830052 [3]广东省中山市坦洲镇农业服务中心,中山528400

出　　处：《植物保护》2023年第1期108-117,共10页Plant Protection

基　　金：广东省基础与应用基础研究基金(2021A1515012502);新疆维吾尔自治区重点研发专项(2022B02033-1);新疆农业科学院科技创新重点培育专项(xjkcpy-2020004)。

摘　　要：本文研究了东革内酯抑制斜纹夜蛾卵巢细胞(SL-221)增殖及凋亡的机理。首先,采用CCK-8法检测东革内酯对细胞增殖的抑制率;其次,利用流式细胞术测定东革内酯对SL-221细胞周期、凋亡和线粒体膜电位的影响;最后,通过RT-qPCR技术检测凋亡相关基因SL-p53、SL-Cytochrome C、SL-InR、SL-Bcl-2、SL-mTOR和SL-PI3K的mRNA表达情况。结果表明:东革内酯对SL-221细胞有明显的增殖抑制活性,且与浓度呈正相关,48 h的IC50为1.98μg/mL;流式细胞术检测到东革内酯可将SL-221细胞周期阻滞于G2/M期,降低线粒体膜电位,同时诱导细胞凋亡;RT-qPCR检测发现东革内酯可诱导凋亡标志基因SL-p53、SL-InR和SL-Cytochrome C的表达显著上调(P<0.05),同时抑制凋亡抑制因子SL-Bcl-2的表达,而细胞凋亡信号通路上游的SL-mTOR和SL-PI3K基因表达均显著下调(P<0.05)。综上,东革内酯可显著抑制SL-221细胞的增殖活性,并通过InR-PI3K-mTOR-Bcl-2信号通路诱导其凋亡。To investigate the mechanism of eurycomalactone inhibiting proliferation and inducing apoptosis in Spodoptera litura ovarian cells(SL-221),the inhibition rate of cell proliferation was detected using CCK-8 kit,and the effects of eurycomalactone on SL-221 cell cycle,apoptosis and mitochondrial membrane potential were determined by flow cytometry.The mRNA expression of apoptosis-related genes SL-p53,SL-Cytochrome C,SL-InR,SL-Bcl-2,SL-mTOR and SL-PI3K were analyzed by RT-qPCR.The results showed that eurycomalactone had obvious proliferation inhibition activity on SL-221 cells,and it was positively correlated with the concentration.The IC50value was 1.98μg/mL when cells were treated for 48 h.Flow cytometry showed that eurycomalactone could block the SL-221 cell cycle at the G2/M phase,reduce mitochondrial membrane potential,and induce apoptosis at the same time.RT-qPCR showed that eurycomalactone could induce significant up-regulation of apoptosis marker genes SL-p53,SL-InR and SL-Cytochrome C(P<0.05),while inhibiting the expression of the apoptosis inhibitor SL-Bcl-2.However,the expressions of SL-mTOR and SL-PI3K genes upstream of the apoptosis pathway were significantly down-regulated(P<0.05).In conclusion,eurycomalactone could significantly inhibit the proliferation activity of SL-221 cells and induce apoptosis through the InR-PI3K-mTOR-Bcl-2 signaling pathway.

关 键 词：东革内酯 斜纹夜蛾 细胞增殖抑制率 细胞凋亡 

分 类 号：S435[农业科学—农业昆虫与害虫防治]