蜗牛多肽混合物抗过氧化氢诱导的SH-SY5Y细胞损伤的作用

Snail polypeptide mixtures protect against damage of SH-SY5Y cells induced by hydrogen peroxide

目的观察蜗牛多肽混合物对过氧化氢(H2O2)诱导SH-SY5Y细胞氧化损伤的抑制作用及其可能机制。方法用H2O2诱导SH-SY5Y细胞损伤,MTT法测定细胞存活率;39～1250 mg·L-1蜗牛多肽混合物处理后,Hoechst染色检测细胞凋亡;罗丹明123染色检测线粒体通透性;细胞免疫化学技术和Western blot技术分别检测增殖细胞核抗原(PCNA)和脑源性神经营养因子(BDNF)的表达。结果 1.54 mmol·L-1H2O2可诱导SH-SY5Y细胞凋亡。用39～156 mg·L-1浓度的蜗牛多肽混合物处理后,H2O2诱导的SH-SY5Y细胞凋亡数量明显减少,同时可减少H2O2引起的线粒体通透性增加,升高PCNA与BDNF的表达(P﹤0.01)。结论蜗牛多肽混合物可抑制过氧化氢诱导的SH-SY5Y细胞凋亡,其作用机制可能与其增加细胞PCNA与BDNF的表达有关。

Objective To study the repressive effect of snail polypeptide mixtures （SPM） on oxidative injury in SH -SY5Y cells induced by hy-drogen peroxide （ H2 02 ）. Methods Hydrogen peroxide induces SH - SY5Y cell damage. The viability of SH - SY5Y cells was measured with MTT assay. Morphological change of cell nucleus were detected with Ho-echst staining. Mitochondrial membrane potential （MMP） were detected with Rhodamine123 staining. The expression of proliferating cell nuclear antigen（PCNA） and brain derived neurotrophic factor（BDNF） were de-termined with immunocytochemistry and western blotting. Results The concentration of 1.54 mmol · L^-1 H2 02 could significantly induced oxi-dative injury in SH - SY5Y cells, which leads to nuclear condensation, MMP dissipation and decrease of PCNA and BDNF expression. In the presence of 39 mg · L^-1 and 156 mg · L^-1 SPM, hydrogen peroxide-in-duced morphological change of nucleus and MMP dissipation were attenu-ated. With further immunocytoehemical analysis, up-regulated PCNA and BDNF expression were observed （P 〈 0. 01 ）. Conclusion SPM could protect against H2O2 -induced SH-SY5Y cells apoptosis, which mechanism correlated with PCNA and BDNF up-regulation.