【摘要】 目的研究微囊藻毒素-LR(microcystins-LR,MC-LR)对人支气管上皮细胞(16HBE)凋亡及凋亡相关蛋白表达的影响。方法将16HBE细胞分别暴露于终浓度为0(对照)、1、10、20、30、40μg/ml的MC-LR溶液24h后,采用MTT法检测细胞活性。将16HBE细胞分别暴露于终浓度为0(对照)、2.5、5、10μg/ml的MC-LR溶液培养24、48 h后,采用流式细胞术检测细胞凋亡率,采用Western blot法检测凋亡相关蛋白细胞色素C(Cyt-c)、B淋巴细胞瘤-2蛋白(Bcl-2)、前凋亡蛋白(Bax)的相对表达量。结果与对照组相比,各浓度MC-LR染毒组16HBE细胞的存活率较低,凋亡率较高,差异均有统计学意义(P<0.05);随着MC-LR染毒浓度的升高,16HBE细胞的存活率呈逐渐下降的趋势,凋亡率呈上升趋势;随着MC-LR染毒时间的延长,16HBE细胞的凋亡率均呈上升趋势。当暴露时间一定时,与对照组相比,各浓度MC-LR染毒组16HBE细胞Bax和Cyt-c蛋白的表达水平均升高,Bcl-2蛋白的表达水平均下降(除外2.5μg/ml MC-LR染毒24 h组),差异有统计学意义(P<0.05);且随着MC-LR染毒浓度的升高和染毒时间的延长,16HBE细胞Bax和Cyt-c蛋白的表达水平均呈上升趋势,Bcl-2蛋白的表达水平均呈下降趋势。结论 MC-LR能诱导16HBE细胞发生凋亡,并引起Cyt-c和Bax蛋白表达水平升高,Bcl-2蛋白表达水平降低。更多还原

【Abstract】 Objective To observe the effects of microcystin-LR(MC-LR) on apoptosis and expression of apoptosis-related proteins in human bronchial epithelial cells(16HBE). Methods Cell viability was assessed by MTT after treatment by MC-LR with different concentration(0-40 μg/ml) for 24 h. After treatment with MC-LR(0,2.5,5,10 μg/ml) for 24 h and 48 h,rate of apoptosis was measured by Annexin V- FITC and PI combined with flow cytometry. The expression of cytochrome-c(Cyt-c),B-cell lymphoma-2(Bcl-2) and Bcl-2 associated X protein(Bax) were detected with Western blot. Results Cells viability decreased with the increase of MC-LR concentration. Apoptosis rates of 16 HBE cells increased with the increase of MC-LR concentration during a certain exposure period,increased with the increase of MC-LR exposure time at a certain concentration.the relative expression of Cyt-c,Bax in 16 HBE cells increased and the expression of Bcl-2 decreased in the groups of 5 μg/ml and 10 μg/ml MC-LR compared with those of control group with statistical significance(P<0.05). Conclusion Microcystin-LR induces apoptosis of 16 HBE,increases the expression of Bax and Cyt-c,and reduce the expression of Bcl-2.更多还原