title: Azadirachtin-induced apoptosis involves lysosomal membrane permeabilization and cathepsin L release in Spodoptera frugiperda Sf9 cells.
abstract: Azadirachtin as a kind of botanical insecticide has been widely used in pest control. We previously reported that azadirachtin could induce apoptosis of Spodoptera litura cultured cell line Sl-1, which involves in the up-regulation of P53 protein. However, the detailed mechanism of azadirachtin-induced apoptosis is not clearly understood in insect cultured cells. The aim of the present study was to address the involvement of lysosome and lysosomal protease in azadirachtin-induced apoptosis in Sf9 cells. The result confirmed that azadirachtin indeed inhibited proliferation and induced apoptosis. The lysosomes were divided into different types as time-dependent manner, which suggested that changes of lysosomes were necessarily physiological processes in azadirachtin-induced apoptosis in Sf9 cells. Interestingly, we noticed that azadirachtin could trigger lysosomal membrane permeabilization and cathepsin L releasing to cytosol. Z-FF-FMK (a cathepsin L inhibitor), but not CA-074me (a cathepsin B inhibitor), could effectively hinder the apoptosis induced by azadirachtin in Sf9 cells. Meanwhile, the activity of caspase-3 could also be inactivated by the inhibition of cathepsin L enzymatic activity induced by Z-FF-FMK. Taken together, our findings suggest that azadirachtin could induce apoptosis in Sf9 cells in a lysosomal pathway, and cathepsin L plays a pro-apoptosis role in this process through releasing to cytosol and activating caspase-3.
authors: Key Laboratory of Natural Pesticide and Chemical Biology, Ministry of Education, and Lab of Insect Toxicology, South China Agricultural University, No. 483, Wushan, Tianhe, Guangzhou 510642, People's Republic of China.
authors: Zheng
authors: Z
authors: Wang
authors: Key Laboratory of Natural Pesticide and Chemical Biology, Ministry of Education, and Lab of Insect Toxicology, South China Agricultural University, No. 483, Wushan, Tianhe, Guangzhou 510642, People's Republic of China; Institute of Natural Product Chemistry, Zhongkai University of Agriculture and Engineering, Guangzhou 510225, People's Republic of China. Electronic address: anzai_28@163.com.
authors: Xingan
authors: X
authors: Cheng
authors: Key Laboratory of Natural Pesticide and Chemical Biology, Ministry of Education, and Lab of Insect Toxicology, South China Agricultural University, No. 483, Wushan, Tianhe, Guangzhou 510642, People's Republic of China.
authors: Qianqian
authors: Q
authors: Meng
authors: Key Laboratory of Natural Pesticide and Chemical Biology, Ministry of Education, and Lab of Insect Toxicology, South China Agricultural University, No. 483, Wushan, Tianhe, Guangzhou 510642, People's Republic of China.
authors: Peidan
authors: P
authors: Wang
authors: Key Laboratory of Natural Pesticide and Chemical Biology, Ministry of Education, and Lab of Insect Toxicology, South China Agricultural University, No. 483, Wushan, Tianhe, Guangzhou 510642, People's Republic of China.
authors: Benshui
authors: B
authors: Shu
authors: Key Laboratory of Natural Pesticide and Chemical Biology, Ministry of Education, and Lab of Insect Toxicology, South China Agricultural University, No. 483, Wushan, Tianhe, Guangzhou 510642, People's Republic of China.
authors: Qiongbo
authors: Q
authors: Hu
authors: Key Laboratory of Natural Pesticide and Chemical Biology, Ministry of Education, and Lab of Insect Toxicology, South China Agricultural University, No. 483, Wushan, Tianhe, Guangzhou 510642, People's Republic of China. Electronic address: humy@scau.edu.cn.
authors: Meiying
authors: M
authors: Hu
authors: Key Laboratory of Natural Pesticide and Chemical Biology, Ministry of Education, and Lab of Insect Toxicology, South China Agricultural University, No. 483, Wushan, Tianhe, Guangzhou 510642, People's Republic of China. Electronic address: guohuazhong@scau.edu.cn.
authors: Guohua
authors: G
authors: Zhong
mesh_terms: D000818:Animals; D017209:Apoptosis / Q000187:drug effects*; D002401:Cathepsin B / Q000378:metabolism; D056668:Cathepsin L / Q000378:metabolism*; D002463:Cell Membrane Permeability / Q000187:drug effects; D049109:Cell Proliferation; D019476:Insect Proteins / Q000378:metabolism*; D007306:Insecticides / Q000494:pharmacology*; D036701:Limonins / Q000494:pharmacology*; D008247:Lysosomes / Q000187:drug effects / Q000378:metabolism*; D061987:Sf9 Cells; D015398:Signal Transduction; D018411:Spodoptera
chemical_list: D019476:Insect Proteins; D007306:Insecticides; D036701:Limonins; D002401:Cathepsin B; D056668:Cathepsin L; C010329:azadirachtin
keywords: Apoptosis; Azadirachtin; Caspase-3; Cathepsins; Lysosomal membrane permeabilization; Spodoptera frugiperda