›› 2007, Vol. 50 ›› Issue (5): 481-487.

• 研究论文 • 上一篇    下一篇

胡椒碱对白纹伊蚊C6/36细胞株的毒性作用

王玉芳,诸葛洪祥*, 周霞, 黄利红   

  1. (苏州大学医学院,江苏苏州 215000)
  • 出版日期:2007-06-08 发布日期:2007-10-20
  • 通讯作者: 诸葛洪祥

Cytotoxicity and apoptosis on C6/36 cell produced by piperine

WANG Yu-Fang, ZHUGE Hong-Xiang, ZHOU Xia, HUANG Li-Hong   

  1. (Medical College of Suzhou University, Suzhou, Jiangsu 215000, China)
  • Online:2007-06-08 Published:2007-10-20
  • Contact: ZHUGE Hong-Xiang

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

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 IC50 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.

Key words: Aedes albopictus, piperine, C6/36 cell line, apoptosis