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丁烯氟虫腈是在农业害虫防治中应用广泛的苯基吡唑类杀虫剂,以家蚕卵巢培养细胞(BmN)为材料,研究丁烯氟虫腈对家蚕的毒性及其作用机制。用MTT法检测丁烯氟虫腈对BmN细胞的增殖有较强抑制作用,且在药液质量浓度12.5~800μg/mL范围内,其抑制作用呈现出浓度依赖性,200、400μg/mL时抑制率分别高达41.34%±7.30%、56.40%±6.70%。用400μg/mL丁烯氟虫腈药液处理BmN细胞48 h后,用流式细胞仪检测发现可导致DNA合成期(S期)细胞比率显著提高(由33.51%±5.12%增加至44.33%±3.98%),DNA合成前期(G0/G1)、DNA合成后期(G2/M)细胞比率降低(分别由37.31%±5.71%、29.18%±7.65%降至31.65%±2.33%、24.02%±3.66%),细胞早期凋亡率和晚期凋亡率分别由4.74%±1.29%、4.95%±1.46%增加至9.86%±2.51%、25.37%±3.38%;用qRT-PCR法检测细胞中4种半胱氨酸蛋白酶(Caspase-1、Caspase-3、Caspase-4、Caspase-5A)基因mRNA转录水平显著上调。研究结果提示,丁烯氟虫腈可能通过上调BmN细胞的半胱氨酸蛋白酶基因表达水平,诱导BmN细胞S期阻滞和凋亡,从而抑制BmN细胞增殖。
Butene fipronil is a widely used phenylpyrazole insecticide in agricultural pest control. Bombyx mori ovary cultured cells (BmN) were used as materials to study the toxicity of buprenorphine to silkworm and its mechanism. The MTT method was used to detect the inhibitory effect of buprenorphine on the proliferation of BmN cells in a dose-dependent manner in the concentration range of 12.5-800 μg / mL. The inhibition rate was 200,400 μg / mL Rates were as high as 41.34% ± 7.30%, 56.40% ± 6.70%. BmN cells treated with 400μg / mL butene fipronil for 48 h showed that the percentage of cells in the DNA synthesis phase (S phase) increased significantly from 33.51% ± 5.12% to 44.33% ± The percentage of cells in the early stage of DNA synthesis (G0 / G1) and late stage of DNA synthesis (G2 / M) decreased from 37.31% ± 5.71%, 29.18% ± 7.65% to 31.65% ± 2.33%, 24.02% ± 3.66 %). The rates of early apoptosis and late apoptosis were increased from 4.74% ± 1.29% and 4.95% ± 1.46% to 9.86% ± 2.51% and 25.37% ± 3.38%, respectively. Four kinds of cells were detected by qRT-PCR Caspase-1, Caspase-3, Caspase-4, Caspase-5A gene mRNA transcription levels were significantly up-regulated. The results suggest that buprenorphine may inhibit the proliferation of BmN cells by up-regulating the expression of cysteine protease gene in BmN cells and inducing S phase arrest and apoptosis in BmN cells.