丙戊基褪黑素与褪黑素对花萼海绵诱癌素所致神经毒性的影响

Effect of Melatonylvalpromide and Melatonin on Calyculin A Induced Neurotoxicity

关键词:  丙戊基褪黑素  褪黑素  花萼海绵诱癌素  神经毒性

Key words:  melatonylvalpromide  melatonin  calyculin A  neurotoxicity

摘要:

目的 观察丙戊基褪黑素(melatonylvalpromide，Mtv)与褪黑素(melatonin，Mt)对花萼海绵诱癌素(calyculin A，CA)所致神经毒性的影响。方法 分别给予DMSO，5 nmol·L^-1 CA，5 nmol·L^-1 CA和50 μmol·L^-1 Mt，5 nmol·L^-1 CA和50 μmol·L^-1 Mtv处理N2a细胞12 h，倒置相差显微镜下观察细胞的形态，Hoechst33342染色后用荧光显微镜摄像观测细胞凋亡率，TBA法测定细胞内丙二醛(MDA)含量，联苯三酚自氧化法测定超氧化物歧化酶(SOD)活性，改良比色法测定过氧化氢酶(CAT)活性。结果 CA处理使N2a细胞突起显著变短变细，细胞凋亡率显著增加，细胞内MDA含量升高，SOD和CAT活性降低。Mt和Mtv均可减轻CA引起的细胞突起缩短和细胞凋亡，也能部分对抗CA引起的SOD和CAT活性抑制以及细胞内MDA含量升高，Mtv的作用强于Mt。结论 Mtv对CA所致神经毒性的保护作用强于Mt。

Abstract:

OBJECTIVE To investigate the effect of melatonylvalpromide(Mtv) or melatonin(Mt) on calyculin A(CA) induced neurotoxicity. METHODS N2a cells were treated with DMSO, or 5 nmol·L^-1 CA, or 5 nmol·L^-1 CA and 50 μmol·L^-1 Mt, or 5 nmol·L^-1 CA and 50 μmol·L^-1 Mtv. Morphology changes were observed with inverted phase contrast microscope, index of apoptotic cell were measured with Hoechst33342 staining, MDA level of cell was analyzed with TBA assay, SOD activity was determined by assessing the inhibition of pyrogallol autooxidation, and CAT activity was analyzed with Colorimetry. RESULTS CA treatment resulted in the process of N2a cell retracting, thinning, and cell apoptosis, accompany with MDA level increasing, SOD and CAT activity decreasing. Both Mt and Mtv could partially antagonize the toxicity of CA, characterized by increasing processes length of cell, decreasing apoptotic cell population, and decreasing MDA level and increasing SOD activity. Moreover, the protective effect of Mtv was stronger than that of Mt. CONCLUSION Mtv has stronger protective effect than that of Mt on CA induced neurotoxicity.