發(fā)布時間：2018-06-01 05:33

  本文選題：丁氟螨酯 + SH-SYY細胞��； 參考：《中國藥理學(xué)與毒理學(xué)雜志》2017年04期

【摘要】：目的研究丁氟螨酯對人經(jīng)神經(jīng)母細胞瘤細胞SH-SY5Y細胞的毒性作用及其機制。方法加入丁氟螨酯0.03,0.06,0.125,0.25,0.5,1,2,2.6,4,6,8和16 mmol·L~(-1)處理SH-SY5Y細胞48 h,MTT法測定細胞存活;DCFH-DA熒光探針標(biāo)記法檢測細胞內(nèi)活性氧(ROS)水平;JC-1標(biāo)記法檢測細胞線粒體膜電位;Hoechst 33258染色觀察細胞核形態(tài);碘化丙啶(PI)染色和流式細胞儀檢測細胞周期和細胞凋亡;Western蛋白印跡法檢測磷酸化P38蛋白(p-P38)和磷酸化Jun激酶(p-JNK)表達水平。結(jié)果與溶劑(DMSO)對照組相比,共孵育48 h后,丁氟螨酯≥0.06 mmol·L~(-1)時可降低細胞存活率(P0.05),且隨濃度增高細胞存活率有降低趨勢,IC_(50)為2.6 mmol·L~(-1);丁氟螨酯1,2,4和6 mmol·L~(-1)組細胞ROS水平升高(P0.01),線粒體膜電位下降(P0.01)。Hoechst 33258染色結(jié)果顯示,丁氟螨酯2,4和6 mmol·L~(-1)組SH-SY5Y細胞出現(xiàn)顆粒狀熒光,細胞核固縮和崩解;流式細胞儀檢測結(jié)果顯示,丁氟螨酯2,4和6 mmol·L~(-1)組細胞凋亡率由DMSO對照組的(0.7±0.1)%分別上升至(6.7±0.1)%,(72.4±8.6)%和(90.7±3.2)%(P0.01);丁氟螨酯4和6 mmol·L~(-1)組G_1期細胞較DMSO對照組明顯增加(P0.01);Western蛋白印跡結(jié)果表明,丁氟螨酯4和6 mmol·L~(-1)組p-JNK表達均升高(P0.01),丁氟螨酯6 mmol·L~(-1)組p-P38表達水平增加(P0.01)。結(jié)論丁氟螨酯可能通過氧化損傷、激活P38蛋白和JNK蛋白誘導(dǎo)SH-SY5Y細胞G_1期阻滯和凋亡。
[Abstract]:Objective to study the toxic effect of bufluride on human transneuroblastoma (SH-SY5Y) cells and its mechanism. Methods SH-SY5Y cells were treated with 0.03 ~ 0.06 ~ (0.125) ~ (0.25) ~ (0.125) ~ 0.25 ~ 0.25 ~ (5) and 16 mmol 路L ~ (-1) ~ (-1) respectively. The viability of SH-SY5Y cells was detected by fluorescence probe labeling method. The level of reactive oxygen species (Ros) was detected by JC-1 labeling method. The cell mitochondria membrane potential was detected by Hoechst 33258 staining and the nuclear morphology was observed by Hoechst 33258 staining. The expression of phosphorylated P38 protein (p-P38) and phosphorylated Jun kinase (p-JNKK) were detected by Western blot and flow cytometry. Results compared with the control group, the control group was incubated for 48 hours. The cell survival rate decreased with the increase of cell concentration, and the cell survival rate decreased with the increase of concentration. The cell ROS level in the groups of bufluroate 鈮，

本文編號：1963027