印楝素诱导细胞凋亡途径的研究

徐 霖, 李 升, 王 嘉福
贵阳学院生物与环境工程学院 贵州大学农业生物工程省重点实验室

摘要


目的：印楝素是一种高效低毒的植物源农药，其作用机制目前仍未清楚，深入研究印楝素诱导细胞凋亡的机制有助于进一步利用印楝素农药；方法：采用基因芯片技术结合实时定量 PCR 技术研究印楝素作用过程中相关基因表达差异变化，流式细胞仪结合 Fluo-3 AM 荧光探针检测 Ca2+ 浓度变化；结果：利用基因芯片技术检测到印楝素处理后，与凋亡途径相关 22个基因差异化表达，其中，钙调蛋白（CaM）基因上调 5.05 倍，是上调最多的基因；利用实时定量 PCR 技术验证了凋亡途径中相关基因，证实印楝素通过触发 Ca2+ 释放诱导凋亡；采用流式细胞仪结合 Fluo-3 AM 荧光探针检测到当 Ca2+ 途径受阻，细胞通过蜕皮激素受体途径诱导凋亡；结论：印楝素通过触发 Ca2+ 释放诱导凋亡，并发现 Ca2+ 通过 Ca2+-CaM 信号途径或蜕皮激素受体 ECR/ USP 级联信号途径诱导凋亡。

关键词


印楝素；实时定量 PCR；Ca2+-CaM 信号途径；蜕皮激素受体；ECR/ USP 级联信号途径

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参考


[1] Radhaw a N.S,Parmar B.S,In: Lalo. P. ed. .Recent Advance in India En-omology[J] . APC Publications,1996: 67-82.

[2] Aritaku L.A,Ramaswamy S,Annadura I．Actin cytoskeleton as a putative target of the neem limonoid Azadirachtin A[J].Insect Bioc hemistry and Molecular Biology ,2007(37) : 627-63.

[3] Kumar CH SSR,Malladi Srinivas,Shirish Yakkundi. Limonoids from the seeds of Azadirachta indica [J]. Phytoche mistry.1996,43(2):451-4557.

[4] Yamasaki K.B,Klocke J.A,Mark Lee S. Isolation and purification of azadirachtin from neem(Azadirachta indica)seeds using flash chromat ography and high performance liquid chromatography[J]. Journal of Chromatography,1986,356:220-226.

[5] 徐霖 , 杨明飞 . 运用 MTT 比色法检测印楝素对果蝇 S2 细胞的毒力作用 [J]. 河南农业大学学报 ,2012,46(1):72-75.

[6] 徐霖 , 冉雪琴 , 王家福 . 印楝素作用分子靶标的研究 [J]. 河南农业大学学报 ,2014,48(1) :83-86.

[7] Giorgi C, Romagnoli A, Pinton P and Rizzuto R, Ca2+ signaling, mitochondria and cell death[J]. Curr Mol Med, 2008, 8(2):119–130.

[8] Wang HG, Pathan N, Ethell IM, Krajewski S, Yamaguchi Y, Shibasaki F, Ca2+-induced apoptosis through calcineurin

dephosphorylation of BAD[J]. Science, 1999, 284:339–343.

[9] Tombal B, Weeraratna AT, Denmeade SR and Isaacs JT, Thapsigargin induces a calmodulin/calcineur-independent

apoptotic cascade responsible for the death of prostatic cancer cells[J]. Prostate, 2000, 43:303–317.

[10] Hajnóczky G, Csordás G, Das S, Garcia-Perez C, Saotome M, Sinha Roy S, Mitochondrial calcium signalling and cell

death: approaches for assessing the role ofmitochondrial Ca2+ uptake in apoptosis[J]. Cell Calcium, 2006, 40:553–560.

[11] Stefan MI, Marshall DP and Le Novère N, Structural analysis and stochastic modelling suggest amechanism for

calmodulin trapping by CaMKII[J]. PLoS ONE 2012, 7:e29406 .

[12] Cakouros D, Daish T, Martin D, Baehrecke EH and Kumar S, Ecdysone-induced expression of the caspase DRONC during hormone-dependent programmed cell death in Drosophila is regulated by Broad-Complex[J]. J Cell Biol, 2002, 157:985–996.

[13] Baehrecke EH, Steroid regulation of programmed cell death during Drosophila development[J]. Cell Death Differ, 2000, 7:1057–1062 .

[14] Adel MM and Sehnal F, Azadirachtin potentiates the action of ecdysteroid agonist RH-2485 in Spodoptera littoralis[J]. J Insect Physiol, 2000, 46:267–274.

[15] Palanker L, Necakov AS, Sampson HM, Ni R, Hu C, Thummel CS. Dynamic regulation of Drosophila nuclear receptor activity in vivo[J]. Development, 2006, 133:3549–3562.




DOI: http://dx.doi.org/10.18686/nykjgl.v2i1.27318

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