title: A COMPREHENSIVE STUDY ON APOPTOSIS INDUCTION BY AZADIRACHTIN IN Spodoptera frugiperda CULTURED CELL LINE Sf9.
abstract: The induction of apoptosis by azadirachtin, a well-known botanical tetranortriterpenoid isolated from the neem tree (Azadirachta indica A. Juss) and other members of the Meliaceae, was investigated in Spodoptera frugiperda cultured cell line (Sf9). Morphological changes in Sf9 cells treated by various concentrations of azadirachtin were observed at different times under light microscopy. Morphological and biochemical analysis indicated that Sf9 cells treated by 1.5 μg/mL azadirachtin showed typical morphological changes, which were indicative of apoptosis and a clear DNA ladder. The flow cytometry analysis showed the apoptosis rate reached a maximum value of 32.66% at 24 h with 1.5 μg/mL azadirachtin in Sf9 cells. The inhibition of Sf9 cell proliferation suggested that the effect of azadirachtin was dose dependent and the EC50 at 48 and 72 h was 2.727 × 10(-6) and 6.348 × 10(-9) μg/mL, respectively. The treatment of azadirachtin in Sf9 cells could significantly increase the activity of Sf caspase-1, but showed no effect on the activity of Topo I, suggesting that the apoptosis induced by azadirachtinin Sf9 cells is through caspase-dependent pathway. These results provided not only a series of morphological, biochemical, and toxicological comprehensive evidences for induction of apoptosis by azadirachtin, but also a reference model for screening insect cell apoptosis inducers from natural compounds.
authors.affiliation: Laboratory of Insect Toxicology, Key Laboratory of Natural Pesticide and Chemical Biology, Ministry of Education, South China Agricultural University, Guangzhou, People's Republic of China.
authors.forename: Benshui
authors.initials: B
authors.lastname: Shu
authors.affiliation: Laboratory of Insect Toxicology, Key Laboratory of Natural Pesticide and Chemical Biology, Ministry of Education, South China Agricultural University, Guangzhou, People's Republic of China.
authors.forename: Wenxiang
authors.initials: W
authors.lastname: Wang
authors.affiliation: Laboratory of Insect Toxicology, Key Laboratory of Natural Pesticide and Chemical Biology, Ministry of Education, South China Agricultural University, Guangzhou, People's Republic of China.
authors.forename: Qingbo
authors.initials: Q
authors.lastname: Hu
authors.affiliation: Laboratory of Insect Toxicology, Key Laboratory of Natural Pesticide and Chemical Biology, Ministry of Education, South China Agricultural University, Guangzhou, People's Republic of China.
authors.forename: Jingfei
authors.initials: J
authors.lastname: Huang
authors.affiliation: Laboratory of Insect Toxicology, Key Laboratory of Natural Pesticide and Chemical Biology, Ministry of Education, South China Agricultural University, Guangzhou, People's Republic of China.
authors.forename: Meiying
authors.initials: M
authors.lastname: Hu
authors.affiliation: Laboratory of Insect Toxicology, Key Laboratory of Natural Pesticide and Chemical Biology, Ministry of Education, South China Agricultural University, Guangzhou, People's Republic of China.
authors.forename: Guohua
authors.initials: G
authors.lastname: Zhong
mesh_terms: D000818:Animals; D017209:Apoptosis / Q000187:drug effects*; D020170:Caspase 1 / Q000378:metabolism; D049109:Cell Proliferation / Q000187:drug effects; D053938:DNA Fragmentation / Q000187:drug effects; D004264:DNA Topoisomerases, Type I / Q000378:metabolism; D005434:Flow Cytometry; D007306:Insecticides / Q000494:pharmacology*; D036701:Limonins / Q000494:pharmacology*; D061987:Sf9 Cells; D018411:Spodoptera / Q000166:cytology
chemical_list: D007306:Insecticides; D036701:Limonins; D020170:Caspase 1; D004264:DNA Topoisomerases, Type I; C010329:azadirachtin
keywords: Sf caspase-1; Sf9; apoptosis; azadirachtin