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目的:观察栀子苷和美满霉素联合应用对H_2O_2诱导PC12细胞凋亡的作用,并对其机制进行初步探讨。方法:处于对数生长期的PC12细胞随机分为对照组、模型组、栀子苷组、美满霉素组以及栀子苷/美满霉素联合应用组。用300μmol/L H_2O_2诱导6 h建立PC12细胞凋亡模型。MTT法测定细胞存活率,比色法测定乳酸脱氢酶(LDH)活性,倒置相差显微镜观察细胞形态,Hoechest 33258荧光核染色观察细胞凋亡状况,Annexin V-FITC/PI双染法流式细胞仪检测细胞凋亡率,Western Blot测定Caspase-3蛋白表达。结果:与模型组相比,栀子苷组和美满霉素组PC12细胞存活率提高,LDH释放量减少,细胞凋亡率降低,同时Caspase-3蛋白表达减少(P均<0.05);与栀子苷组和美满霉素组相比,栀子苷/美满霉素联合应用组PC12细胞的存活率进一步提高,LDH释放量进一步减少,细胞凋亡率进一步降低,同时Caspase-3蛋白表达进一步减少(P<0.05或P<0.01)。以上指标相比,差异均有统计学意义。结论:栀子苷与美满霉素联合应用较单一应用能更有效拮抗PC12细胞凋亡,其机制可能与抑制Caspase-3蛋白表达有关。
Objective: To observe the effect of geniposide and minocycline on the apoptosis of PC12 cells induced by H 2 O 2, and to explore its mechanism. Methods: PC12 cells in logarithmic growth phase were randomly divided into control group, model group, geniposide group, minocycline group and geniposide / minocycline combination group. Apoptosis model of PC12 cells was induced by 300μmol / L H 2 O 2 for 6 h. Cell viability was measured by MTT assay. Lactate dehydrogenase (LDH) activity was assayed by colorimetric assay. Cell morphology was observed by inverted phase contrast microscope. Hoechst 33258 fluorescence staining was used to observe apoptotic cells. Annexin V-FITC / PI double staining flow cytometry The rate of apoptosis was detected by Western Blot, and the expression of Caspase-3 protein was detected by Western Blot. Results: Compared with the model group, PC12 cells in geniposide group and minocycline group had higher survival rate, lower LDH release, decreased apoptosis rate and decreased Caspase-3 protein expression (P <0.05) Compared with minocycline group, the combination of geniposide and minocycline increased the survival rate of PC12 cells, further reduced the amount of LDH release and further reduced the apoptosis rate, and further reduced the expression of Caspase-3 protein (P <0.05 or P <0.01). The above indicators, the differences were statistically significant. Conclusion: Geniposide combined with minocycline can antagonize the apoptosis of PC12 cells more effectively than single application, which may be related to the inhibition of Caspase-3 protein expression.