石杉碱甲衍生物 M3 对硝普钠诱导的 PC12 细胞凋亡的保护作用。
Huperzine A derivative M3 protects PC12 cells against sodium nitroprusside-induced apoptosis.
机构信息
State Key Laboratory of Bioactive Substances and Functions of Natural Medicines, Institute of Materia Medica, Chinese Academy of Medical Sciences, Beijing, China.
出版信息
Acta Pharmacol Sin. 2012 Jan;33(1):34-40. doi: 10.1038/aps.2011.147. Epub 2011 Nov 28.
AIM
To investigate the effects of M3, a derivative of huperzine A, on the apoptosis induced by sodium nitroprusside (SNP) in PC12 cells.
METHODS
Cell viability was detected using MTT method. Apoptosis was examined with annexin V/prodium iodide (PI) stain. The levels of reactive oxygen species (ROS) were measured using fluorophotometric quantitation. The amount of malonaldehyde (MDA) was determined with MDA detection kits. The expression of caspase-3 and Hsp70 were analyzed using Western blotting.
RESULTS
Exposure of PC12 cells to SNP (200 μmol/L) for 24 h decreased the cell viability to 69.0% of that in the control group. Pretreatment with M3 (10 μmol/L) or huperzine A (10 μmol/L) significantly protected the cells against SNP-induced injury and apoptosis; the ratio of apoptotic bodies in PC12 cells was decreased from 27.3% to 15.0%. Pretreatment with M3 (10 μmol/L) significantly decreased ROS and MDA levels, and increased the expression of Hsp70 in the cells. Quercetin (10 μmol/L) blocked the protective effect of M3, while did not influence on that of huperzine A.
CONCLUSION
M3 protects PC12 cells against SNP-induced apoptosis, possible due to ROS scavenging and Hsp70 induction.
目的
研究石杉碱甲类似物 M3 对硝普钠（SNP）诱导的 PC12 细胞凋亡的影响。
方法
采用 MTT 法检测细胞活力，用 Annexin V/碘化丙啶（PI）染色法检测细胞凋亡，荧光光度法定量检测活性氧（ROS）水平，MDA 检测试剂盒测定丙二醛（MDA）含量，Western blot 法分析 caspase-3 和 Hsp70 的表达。
结果
SNP（200 μmol/L）孵育 24 h 后，PC12 细胞活力降至对照组的 69.0%。用 M3（10 μmol/L）或石杉碱甲（10 μmol/L）预处理可显著减轻 SNP 诱导的细胞损伤和凋亡，使凋亡小体比例从 27.3%降至 15.0%。M3（10 μmol/L）预处理可显著降低 ROS 和 MDA 水平，增加细胞内 Hsp70 的表达。槲皮素（10 μmol/L）阻断了 M3 的保护作用，但对石杉碱甲的作用无影响。
结论
M3 可保护 PC12 细胞免受 SNP 诱导的凋亡，其机制可能与清除 ROS 和诱导 Hsp70 有关。
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