目的 研究新生鼠常压窒息后脑组织内神经元凋亡与Omi/HtrA2 表达变化及丹参干预的影响。方法 选用新生大鼠常压窒息模型, 7日龄新生SD大鼠随机分为对照组、窒息组和丹参干预组, 于造模后不同时间点(6、12、24、48、72 h)取脑组织行石蜡切片, TUNEL(Terminal dexynucleotidyl transferase-mediated dUTP nick end labeling)(末端脱氧核苷酸转移酶介导的dUTP缺口末端标记测定法)法检测神经细胞凋亡数量, 免疫组织化学方法 检测Omi/HtrA2蛋白表达。结果 窒息组Omi/HtrA2的表达和神经细胞凋亡数量在各时间点均高于对照组(P均<0.05), 窒息组Omi/HtrA2的表达在造模后12 h达峰(IOD10.98±1.34), 而神经元凋亡数量在24 h达峰(12.1±2.66)。与窒息组比较, 丹参干预组各时间点神经细胞凋亡数量和Omi/HtrA2表达均显著下降, 其差异有统计学意义(P均<0.05), 但均未恢复至对照组水平(P<0.05)。结论 常压窒息可能通过上调Omi/HtrA2表达而诱导神经元凋亡, 丹参可通过抑制脑组织内Omi/HtrA2表达而减少神经元凋亡数量, 从而对窒息脑损伤起到保护作用。
Abstract
Objective To study the cell apoptosis and expression of Omi/HtrA2 in brain of neonatal rats after asphyxia and to evaluate the effects of Salvia miltorrhiza bunge( SMB ) on it. Methods A total of 75 SD rats aged 7 days were randomly divided into the following groupscontrol group, asphyxia group and SMB treatment group, then the brain tissues were taken from the rats in the three groups at 6, 12, 24, 48 hours and 72 hours after the asphyxia.The apoptotic cells were detected by the TUNEL staining method, The Omi/HtrA2 protein expression was detected with the immunhistochemistry method. Results The number of apoptotic cells and the expression of Omi/HtrA2 were higher in asphyxia groups than those in control group (P<0.05).In asphyxia groups, the expression of Omi/HtrA2 reached peak at 12 h after asphyxia(10.98±1.34), and the number of apoptotic cells reached peak at 24 h after asphyxia (12.1±2.66).But compared with asphyxia group, the expression of Omi/HtrA2 and the number of apoptotic cells were significantly decreased in SMB treatment group, and the differences were significant(P＜0.05), but all parameters were not restored to the levels of control group(P＜0.05). Conclusion Asphyxia can induce the apoptosis of neuronal, Omi/HtrA2 may be involved in this process.SMB can reduce the number of apoptotic cells by inhibiting the expression of Omi/HtrA2 in brain, and thus it can play a protective role in asphyxia brain damage of neonatal rats.
关键词
窒息 /
新生大鼠 /
神经元 /
凋亡 /
Omi/HtrA2 /
丹参
Key words
asphyxia /
neonatal rat /
neuron /
apoptosis /
Omi/HtrA2 /
Salvia miltiorrhiza Bunge
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基金
四川省卫生厅科研课题(110332);泸州医学院青年基金资助项目(2010)
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