快速诱导细胞凋亡及检测方法研究

Study on Methods of Measuring and Inducing Apoptosis Rapidly

目的:探讨Ca2+和Na+诱导细胞凋亡的最佳浓度及时间,并用甲基绿-派诺宁染色法检测凋亡细胞的形态变化。方法:分别用不同浓度梯度及时间梯度的Ca2+和Na+胁迫处理洋葱鳞茎内表皮细胞,得诱导的最佳浓度及时间;用甲基绿-派诺宁染色法检测诱导凋亡的洋葱鳞茎内表皮细胞、大蒜根尖细胞和鸡血红细胞的形态特征变化。结果:诱导处理的最佳Ca2+和Na+浓度为0.4 mol/L,最适时间约为8 h,且CaCl2的诱导效果较NaCl好;经甲基绿-派诺宁染色,洋葱鳞茎内表皮细胞、大蒜根尖细胞、鸡血红细胞凋亡细胞的细胞核均呈蓝紫色,细胞质呈红色。结论:找出了诱导细胞凋亡的最适Ca2+和Na+浓度和时间,并检测到细胞凋亡。

Objective： To study the optimum concentration and time under Ca2＋ and Na＋, and detect the chang- es of apoptosis morphological characteristics by methyl green-pyronin staining. Methods： To get the optimum con- centration and the optimum time, onion skin cells within the bulb were observed and counted under the micro- scope after different concentration and different time of ions stress treatment. The methyl green-pyronin staining was used to detect the changes of induced apoptosis morphological characteristics for onion skin cells within the bulb, garlic root cells and chicken lurid cells. Results： The optimal concentration of Ca2＋ and Na＋ were 0.4 mol/ L, the optimal induced time was 8 h, and the effect of CaCl2 was better than that of NaC1. Using methyl green and pyronin staining, we could see that the nucleus of onion skin apoptosis cells within the bulb, garlic root cells and chicken lurid apoptosis cells were dyed blue-violet, and their cytoplasm were red. Conclusion： The optimal ionic stress concentration and the best time were obtained, and the apoptosis was successfully detected.