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胡椒碱对白纹伊蚊C6/36细胞株的毒性作用
引用本文:王玉芳,诸葛洪祥,周霞,黄利红.胡椒碱对白纹伊蚊C6/36细胞株的毒性作用[J].昆虫学报,2007,50(5):481-487.
作者姓名:王玉芳  诸葛洪祥  周霞  黄利红
作者单位:(苏州大学医学院,江苏苏州 215000)
基金项目:国家自然科学基金项目(39970654)
摘    要:采用MTT检测法、Annexin-V/PI双染流式细胞仪检测法及细胞形态学观察法,研究了植物型杀虫剂胡椒碱对白纹伊蚊Aedes albopictus C6/36细胞株的细胞毒性、诱导细胞凋亡与坏死的关系、Fas蛋白的表达、细胞生长状态改变及受损细胞的恢复。结果表明:胡椒碱浓度在0.035 mmol/L以上时可以诱发C6/36细胞形态学改变,在倒置显微镜下表现为细胞呈多形性,细胞胀大或皱缩,细胞间隙增宽,大片细胞聚集成团,脱落、崩解、死亡。用胡椒碱处理24 h后,对C6/36细胞的半数毒性浓度(IC50)为 0.32 mmol/L,且毒性作用强度随着药物浓度增加而增强。Annexin-V/PI双染法检测结果显示药物浓度在0.28 mmol/L 以上诱导的细胞凋亡明显,药物浓度为0.56 mmol/L 时细胞凋亡率达19.4%,而药物引起的细胞总死亡率为72.7%;Fas蛋白表达在药物浓度0.28 mmol/L 以上时有所上调,但不明显。高浓度胡椒碱(0.56 mmol/L)分别作用于C6/36细胞24和48 h,在经历一段生长停滞后,24 h组受损细胞均可缓慢恢复,而48 h组细胞则出现不可逆性的损伤。由此认为:胡椒碱对C6/36细胞株有毒性作用,但药物诱导的细胞凋亡在细胞毒性作用机制中不占主导地位;胡椒碱对C6/36细胞的作用有时间依赖性,延长作用时间,药物对细胞的毒性作用增强。

关 键 词:白纹伊蚊  胡椒碱  C6/36细胞株  细胞凋亡  
文章编号:0454-6296(2007)05-0481-07
收稿时间:2006-10-10
修稿时间:2007-03-01

Cytotoxicity and apoptosis on C6/36 cell produced by piperine
WANG Yu-Fang,ZHUGE Hong-Xiang,ZHOU Xia,HUANG Li-Hong.Cytotoxicity and apoptosis on C6/36 cell produced by piperine[J].Acta Entomologica Sinica,2007,50(5):481-487.
Authors:WANG Yu-Fang  ZHUGE Hong-Xiang  ZHOU Xia  HUANG Li-Hong
Institution:(Medical College of Suzhou University, Suzhou, Jiangsu 215000, China)
Abstract:We observed cytotoxicity,apoptosis,the expression of Fas protein,growth state and recovery of damaged cells produced by botanical insecticide piperine on C6/36 cell strain of Aedes albopictus using MTT,Annexin-V/PI double staining and inverted microscope observation.The results indicated that piperine caused the morphological change in the treated cells at the concentration of above 0.035 mmol/L.The cells showed pantomorphia under the inverted microscope.They became swollen or shrunk,and the space between cells was widened.A large number of cells assembled into conglobations,and then ablated,disaggregated and died.The IC_ 50 of C6/36 cell 24 hours after treatment by piperine was 0.32 mmol/L.Moreover,the cytotoxicity increased with the raising of concentration of piperine.The results of Annexin-V/PI double staining showed that the apoptosis was apparent in treatments of the piperine concentration above 0.28 mmol/L,while the rate of apoptosis was 19.4% in the treatement of the piperine concentration of 0.56 mmol/L.The expression of Fas protein increased in the treatments of the piperine concentration above 0.28 mmol/L but not apparently,and the total death rate was 72.7%.Treated with high concentration of piperine(0.56 mmol/L)for 24 hours and 48 hours respectively,following a period of growth retardation,the cells of 24-hour-group could recover slowly while the 48-hour-group showed inconvertibility.The results suggest that there is time dependence of the effect on C6/36 by piperine.Apoptosis,however,is not the main mechanism.The cytotoxicity would be increased with the treating time prolonged.
Keywords:Aedes albopictus  piperine  C6/36 cell line  apoptosis
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