溴氰菊酯对PC12细胞Fas、FasL、TNFR1蛋白及凋亡的影响  

The Effects of Deltamethrin on Fas,FasL,TNFR1 Protein Expre s sion and Apoptosis in PC12 Cells

作　　者：戴中华[1] 石年[1] 刘恭平[1] 韦艳宏[1] 李煌元[1] 陈丹[1] 王斌[1] 

机构地区：[1]华中科技大学同济医学院公卫学院卫生毒理学系,湖北武汉430030

出　　处：《癌变．畸变．突变》2005年第2期97-100,共4页Carcinogenesis,Teratogenesis & Mutagenesis

基　　金：国家自然科学基金项目(No.30371225)

摘　　要：背景与目的 :探讨溴氰菊酯诱导神经细胞凋亡及其神经毒作用机制。 材料与方法 :用免疫细胞化学法定性检测给予不同剂量水平(10 -4mol/L ,10 -5mol/L ,10 -6mol/L)的溴氰菊酯24h后 ,Fas、FasL、TNFR1蛋白在PC12细胞中的表达 ;流式细胞仪定量检测给予溴氰菊酯24h和48h后 ,Fas、FasL、TNFR1蛋白在PC12细胞中的相对表达量阳性细胞率(rateofpositivecells ,RPC)和平均荧光强度(meanfluorescenceintensity,MFI)及PC12细胞的凋亡率。 结果 :①Fas、FasL、TNFR1蛋白在对照组和各剂量组均为阳性表达 ,但镜下未见各蛋白在各组之间的差异表达。流式细胞仪检测发现 :染毒24h后 ,Fas、FasL蛋白在各组的表达没有差异 ,而TNFR1蛋白在中剂量和高剂量组的表达与对照组比较有升高 ;染毒48h后 ,Fas、FasL、TNFR1蛋白在高剂量组表达(MFI分别为42.85±8.4、37.91±1.65、8.09±1.83)与各自对照组比较均有升高。②染毒24h后 ,各剂量组凋亡率没有升高 ;染毒48h后 ,高剂量组凋亡率与对照组凋亡率比较有升高。③染毒48h组细胞凋亡率与溴氰菊酯浓度、Fas、Fasl蛋白含量有直线相关关系。 结论 :溴氰菊酯可能通过死亡受体Fas诱导PC12细胞凋亡。BACKGROUND AND AIM: To explore the potential mechanisms of neural apoptosis induced by deltamethrin(DM). MATERIAL AND METHODS: PC12cells were respectively given with10 -4 mol/L,10 -5 mol/L and10 -6 mol/L deltamethrin.Fas,FasL and TNFR1protein in PC12cells were qualitatively determined by immunocytochemistry at24hours after deltamethrin treatment.Fas,FasL and TNFR1protein were indirectly marked with fluorescein FITC and their relative contents(rate of positive cells,RPC and mean fluorescence intensity,MFI)were quantitatively determined with flow cytometry,meanwhile apoptosis of PC12cells was studied using flow cytometry with PI staining at24hours and48hours after deltamethrin treatment. RESULTS: Immunostaining for Fas,FasL and TNFR1protein was found in the control and deltamethrin treatment PC12cells.But the different expressions of these three proteins were not detected among the control and deltamethrin treatment PC12cells.Twenty-four hours after deltametrin treatment,we discovered that Fas,FasL protein did not altered but TNFR1protein increased in cells with10 -5 mol/L and10 -4 mol/L deltamethrin treatment(each MFI was3.50±0.26,3.74±0.33)opposite to the control(MFI was2.69±0.19)(P<0.05).Forty-eight hours after cells were exposed to deltematrin,Fas,FasL and TNFR1protein increased only in cells with10 -4 mol/L deltamethrin treatment(each MFI was42.85±8.4,37.91±1.65,8.09±1.83)opposite to their own control(each MFI was29.37±3.07,29.71±0.68,5.30±0.33)(P<0.05).Compared with the control,the apoptotic ratio increased only when the cells were treated with10 -4 mol/L deltamethrin for48hours[(0.60±0.08)% vs(2.62±1.37)%].And there are correlations between apoptotic ratio and DM,Fas,Fasl protein. CONCLUSIONS: Apoptosis of PC12cells induced by deltamethrin maybe be mediated by death receptor Fas.

关 键 词：溴氰菊酯 FAS蛋白 FASL蛋白 TNFR1蛋白 凋亡 流式细胞仪 

分 类 号：R994[医药卫生—毒理学]