CAJ | 학술논문

目的：利用Hoechst33342/PI双染法和原位末端标记法(TUNEL染色检测纳米二氧化硅(nm-SiO2)对神经细胞凋亡的影响，比较两种检测方法的优缺点。方法：以体外培养的人神经母细胞瘤SK-N-SH为研究对象，15和30 nm粒径的nm-SiO2(剂量为2.5、5、10μg/mL)分别处理细胞24 h，另设1~5 m SiO2组和溶剂对照组，采用Hoechst33342/PI双染法和TUNEL染色检测各处理组对SK-N-SH细胞凋亡的影响。结果：与对照组相比，两种检测方法分析均显示了nm-SiO2处理组SK-N-SH细胞凋亡率显著增加(P<0.05)，且具有尺寸、剂量依赖性，而微米级SiO2对凋亡的影响不显著(P>0.05)。结论：nm-SiO2能诱导SK-N-SH细胞凋亡。Hoechst33342/PI双染法特异性高，简单易行；TUNEL法灵敏度高，能检测少量的细胞凋亡，但成本较高，两种方法可结合使用以便更加准确的检测神经细胞的凋亡。)μ
목적：이용Hoechst33342/PI쌍염법화원위말단표기법(TUNEL염색검측납미이양화규(nm-SiO2)대신경세포조망적영향，비교량충검측방법적우결점。방법：이체외배양적인신경모세포류SK-N-SH위연구대상，15화30 nm립경적nm-SiO2(제량위2.5、5、10μg/mL)분별처리세포24 h，령설1~5 m SiO2조화용제대조조，채용Hoechst33342/PI쌍염법화TUNEL염색검측각처리조대SK-N-SH세포조망적영향。결과：여대조조상비，량충검측방법분석균현시료nm-SiO2처리조SK-N-SH세포조망솔현저증가(P<0.05)，차구유척촌、제량의뢰성，이미미급SiO2대조망적영향불현저(P>0.05)。결론：nm-SiO2능유도SK-N-SH세포조망。Hoechst33342/PI쌍염법특이성고，간단역행；TUNEL법령민도고，능검측소량적세포조망，단성본교고，량충방법가결합사용이편경가준학적검측신경세포적조망。)μ
OBJECTIVE: To evaluate the effects of nano-silica(nm-SiO2) on apoptosis of neuroblastoma cells,and compare the advantages and disadvantages of Hoechst33342/PI staining and TUNEL assay in apoptosis measurement. METHODS:Cultured human neuroblastoma SK-N-SH cells were treated with nm-SiO2 (15 and 30 nm) and micro-sized SiO2 at different doses (2.5,5 and 10μg/mL) for 24 h. Apoptotic cells were detected by Hoechst33342/PI and TUNEL staining. RESULTS:Compared with normal control,the percentages of total apoptosis in nm-SiO2-treated cells were significantly increased by both methods. CONCLUSION:nm-SiO2 increased apoptosis of SK-N-SH cells in a dose-and size-dependent manner. Hoechst33342/PI is more specific,cheaper and simpler,while TUNEL assay is more sensitive. It is recommended to combine these two methods to assess apoptosis in neuronal cells.