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目的:观察钩藤碱(rhnchophylline,Rhy)对多巴胺(dopamine,DA)诱导NT2细胞凋亡的防护作用.方法:以LDH的漏出率反映细胞的生存率;用TUNEL染色法和DNA琼脂糖凝胶电泳法观察NT2神经元凋亡情况;用Western blotting法测定Bcl-2蛋白表达.结果:Rhy在5和50μmol/L的浓度下能显著地抑制由DA所致的乳酸脱氢酶的漏出,以及明显地提高以PMS试剂转化为指标的生存率(P<0.05,P<0.01);在分化的NT2细胞神经元中,转染bcl-2基因的神经元凋亡率明显低于未经bcl-2基因转染的神经元,而Rhy使DA诱导的转染bcl-2基因神经元和未转染bcl-2基因神经元的凋亡率均明显减少;Rhy能抑制DA所致的DNA降解,但Phy对NT2细胞Bcl-2蛋白表达无明显影响.结论:Rhy能对抗DA诱导的NT2细胞的损伤.
OBJECTIVE: To observe the protective effect of rhnchophylline (Rhy) on dopamine (DA)-induced apoptosis of NT2 cells. METHODS: The survival rate of LDH was reflected by the leakage rate of LDH; the DNA agarose gel was stained by TUNEL staining. Electrophoresis was used to observe the apoptosis of NT2 neurons; Bcl-2 protein expression was measured by Western blotting. Results: Rhy in 5 and 50 μmol/L concentrations can significantly inhibit the leakage of lactate dehydrogenase caused by DA, and The survival rate of PMS reagents was significantly improved (P<0.05, P<0.01). In differentiated NT2 neurons, the apoptosis rate of neurons transfected with bcl-2 gene was significantly lower than that without bcl- 2Gene-transfected neurons, while Rhy caused DA-induced apoptosis in neurons that were transfected with bcl-2 and untransfected bcl-2 genes; Rhy inhibited DA-induced DNA degradation. However, Phy had no significant effect on the expression of Bcl-2 protein in NT2 cells. Conclusion: Rhy can protect against NT-induced injury of NT2 cells.