美金刚对帕金森病SH-SY5Y细胞模型细胞毒性损伤的影响及相关机制研究  

Effect of memantine on cytotoxic injury in SH-SY5Y cell model of Parkinson's disease and its related mechanism

作　　者：王乐 孙雅丽 岳霞[1] WANG Le;SUN Ya-li;YUE Xia(Department of Neurology,Bayannaoer City Hospital,Bayan Nur Inner Mongolia 015000,China)

机构地区：[1]巴彦淖尔市医院神经内科,内蒙古巴彦淖尔015000

出　　处：《临床和实验医学杂志》2022年第7期682-686,共5页Journal of Clinical and Experimental Medicine

基　　金：内蒙古自治区卫生计生科研计划项目(编号:201703259)。

摘　　要：目的分析美金刚对帕金森病SH-SY5Y细胞模型细胞毒性损伤的影响及相关机制研究。方法采用过氧化氢(H_(2)O_(2))诱导氧化损伤建立帕金森病SH-SY5Y细胞毒性损伤模型。将对数生长期的细胞随机分成4组,分别加入等量浓度为10、20μmol/L的美金刚及磷酸盐缓冲液预处理2 h,3个损伤组再加入400μmol/L的H_(2)O_(2)处理24 h。用MTT法检测SH-SY5Y细胞毒性;用Hoechst33342染色检测细胞凋亡情况;用比色法测定乳酸脱氢酶(LDH)释放率、半胱氨酸蛋白酶-3(Caspase-3)相对酶活性;采用酶联免疫吸附试验(ELISA)试剂盒检测丙二醛和超氧化物歧化酶(SOD)表达水平。结果(1)与C组相比,A、B、D组细胞增殖率显著降低,凋亡率显著增加,差异均有统计学意义(P<0.05)。与D组相比,A、B组细胞增殖率显著增加,凋亡率显著降低,且B组较A组增殖率显著增加,凋亡率显著降低,差异均有统计学意义(P<0.05)。(2)干预24~48 h后,与C组相比,A、B、D组细胞LDH释放率显著增加,差异均有统计学意义(P<0.05)。与D组相比,A、B组细胞LDH释放率显著降低,且B组也较A组显著降低,差异均有统计学意义(P<0.05)。(3)干预24~48 h后,与C组相比,A、B、D组细胞Caspase-3相对酶活性显著增加,差异均有统计学意义(P<0.05)。与D组相比,A、B组细胞Caspase-3相对酶活性显著降低,且B组也较A组显著降低,差异均有统计学意义(P<0.05)。(4)干预24~48 h后,与C组相比,A、B、D组细胞丙二醛水平显著增加,SOD水平显著降低,差异均有统计学意义(P<0.05)。与D组相比,A、B组细胞丙二醛水平显著降低,SOD水平显著增加,且B组较A组增丙二醛水平显著降低,SOD水平显著增加,差异均有统计学意义(P<0.05)。结论美金刚可以阻止H_(2)O_(2)诱导的帕金森病SH-SY5Y细胞毒性损伤,其机制可能与抑制LDH的释放、Caspase-3的活性及降低丙二醛表达水平和促进SOD水平增加有关。Objective To analyze the effect of memantine on cytotoxic injury in SH-SY5Y cell model of Parkinson's disease and its related mechanism.Methods The SH-SY5Y cell toxic injury model of Parkinson's disease was established by H_(2)O_(2) induced oxidative damage.The cells in logarithmic growth phase were randomly divided into 4 groups,which were pretreated with the same volume of memantine(10,20μmol/L)and PBS for 2 h,and then treated with 400μmol/L H_(2)O_(2) for 24 h.The cytotoxicity of SH-SY5Y cells was detected by MTT assay;apoptosis was detected by Hoechst 33342 staining;the release rate of lactate dehydrogenase(LDH),the relative activity of Caspase-3 were measured by colorimetry;Enzyme-linked immunosorbent assay(ELISA)kit was used to detect the expression levels of malondialdehyde(MDA)and superoxide dismutase(SOD).Results(1)Compared with group C,the cell proliferation rate of A,B,and D groups was significantly reduced,and the apoptosis rate was significantly increased,the differences were statistically significant(P<0.05).Compared with group D,the cell proliferation rate and apoptosis rate of A and B groups were significantly increased,and the cell proliferation rate and apoptosis rate of group B were significantly increased than that of group A,the differences were statistically significant(P<0.05).(2)After 24 to 48 hours of intervention,compared with group C,the cell LDH release rate of groups A,B,and D increased significantly,the differences were statistically significant(P<0.05).Compared with group D,the cell LDH release rate of group A and group B was significantly lower,and group B was also significantly lower than group A,the differences were statistically significant(P<0.05).(3)After 24 to 48 hours of intervention,compared with group C,the relative enzyme activity of Caspase-3 in groups A,B,and D increased significantly,the differences were statistically significant(P<0.05).Compared with group D,the relative enzyme activity of Caspase-3 in group A and B was significantly reduced,and group B was also sig

关 键 词：帕金森病 美金刚 乳酸脱氢酶 半胱氨酸蛋白酶-3 丙二醛 

分 类 号：R749.16[医药卫生—神经病学与精神病学]