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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、72h)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12h达峰(IOD:10.98±1.34),而神经元凋亡数量在24h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。
Objective To investigate the changes of neuronal apoptosis and the expression of Omi / HtrA2 in neonatal rats with atmospheric asphyxia and the effect of Danshen intervention. Methods The neonatal SD rats were randomly divided into control group, asphyxia group and Salvia miltiorrhiza intervention group. The rats were sacrificed at different time points (6, 12, 24, 48, 72 h) TUNEL (terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling) method was used to detect the number of apoptotic neurons. Immunohistochemistry was used to detect the expression of Omi / HtrA2 protein expression. Results The expression of Omi / HtrA2 and the number of neuronal apoptosis in asphyxia group were higher than those in control group at all time points (P <0.05). The expression of Omi / HtrA2 in asphyxia group reached peak at 12h after modeling (IOD: 10.98 ± 1.34 ), While the number of apoptotic neurons peaked at 12 h (12.1 ± 2.66). Compared with asphyxia group, the number of apoptotic neurons and the expression of Omi / HtrA2 in Salvia miltiorrhiza intervention group at each time point were significantly decreased (P <0.05), but not recovered to the control group (P <0.05) ). CONCLUSIONS: Aspirin may induce neuronal apoptosis by up-regulating the expression of Omi / HtrA2. Salvia miltiorrhiza can reduce the number of neuronal apoptosis by inhibiting the expression of Omi / HtrA2 in brain tissue, which may play a protective role in asphyxial brain injury.