4种交链孢毒素对人食管上皮细胞Het－1 A的体外毒性研究

Study on the toxicity of four kinds of Alternaria toxins on human esophageal epithelial cell Het-1 A in vitro

目的研究交链孢酚(AOH)、交链孢酚单甲醚(AME)、交链孢菌酮酸(Te A)和腾毒素(TEN)4种交链孢毒素对人食管上皮细胞Het-1 A的体外急性毒性作用。方法将Het-1 A用不同浓度的4种交链孢毒素处理,分别采用3-(4,5-二甲基噻唑-2)-2,5-二苯基四氮唑溴盐(MTT)法、膜联蛋白V-异硫氰酸荧光素/碘化丙啶(Annexin VFITC/PI)双染法、PI单染法和分光光度法研究其对Het-1 A的增殖抑制、细胞凋亡、周期分布和含半胱氨酸的天冬氨酸蛋白水解酶-3(caspase-3)活性的影响。结果 4种交链孢毒素对Het-1 A的半数抑制浓度(IC--_(50))值分别为54.31、43.38、121.91和141.96μmol/L,均可引起细胞凋亡,并可通过引起G_2-M期比例上升而影响细胞的周期分布。AOH和AME可通过剂量依赖增强caspase-3活性引发细胞凋亡。结论 AOH、AME、Te A和TEN可通过抑制细胞增殖、引起细胞凋亡、诱导G_2-M周期阻滞等对Het-1 A产生急性毒性。

Objective To study the acute cytotoxicity of four kinds of Alternaria toxins alternaiol（ AOH）,alternariol monomethyl ether（ AME）,tenuazonic acid（ Te A） and tentoxin（ TEN） on human esophageal epithelium cell Het-1 A in vitro. Methods Het-1 A was treated with different concentrations of 4 kinds of Alternaria toxins. The effects of 4 kinds of Alternaria toxins on cell proliferations,cell apoptosis,cell cycle distribution and cysteinyl aspartate specific proteinase-3（ capase-3） activity were detected by 3-（ 4,5-dimethyl-2-thiazolyl）-2,5-diphenyl-2-H-tetrazolium bromide（ MTT）,annexin V-fluorescein isothiocyanate/propidium iodide（ Annexin V-FITC/PI） double staining method,PI single staining method and spectrophotometry,respectively. Results The half maximal inhibitory concentration（ IC_（50）） of AOH,AME,Te A and TEN on Het-1 A was 54. 31,43. 38,121. 91 and 141. 96 μmol/L,respectively. The 4 kinds of Alternaria toxins could induce the apoptosis of Het-1 A and also could affect the cell cycle distribution through increasing the percent of G_2-M phase. AOH and AME could enhance the activity of caspase-3 in a positive dose-dependent manner to induce the apoptosis of Het-1 A. Conclusion AOH,AME,Te A and TEN could cause acute cytotoxicity on Het-1 A through inhibiting cell proliferation,inducing cell apoptosis and G_2-M cell cycle arrest etc.