标题：八肽胆囊收缩素对缺氧缺血性细胞损伤模型的干预作用

      作者：刘瑛，周江堡    (攀枝花市妇幼保健院儿科，四川 攀枝花 617000)

      卷次： 2017年28卷2期

      【摘要】 目的 研究八肽胆囊收缩素(CCK-8)对缺氧缺血神经细胞凋亡的干预作用，以及凋亡过程中对神经生长因子(NGF)蛋白及NGF mRNA表达的影响。方法 体外培养新生大鼠大脑皮层神经细胞7 d，作为空白对照组(A组)，用50 μmol/L谷氨酸浸泡处理后建立缺氧缺血细胞损伤模型(B组/模型组)，将CCK-8分别以浓度1×10-6 mol/L(C组)、1×10-7 mol/L (D组)、1×10-8 mol/L (E组)进行干预，作用后继续培养细胞24 h，采用原位末端标记技术检测各组细胞凋亡率、免疫细胞化学检测NGF蛋白表达，原位杂交技术检测NGF mRNA表达。结果 A组细胞有少量凋亡，凋亡率为(8.49±4.54)%，C组和D组细胞凋亡率分别为(3.87±5.64)%、(7.84±4.19)%，较B组的(17.53±6.93)%明显降低，差异均有统计学意义(P<0.05)，而E组凋亡率为(13.27±3.41)%，与B组比较差异无统计学意义(P>0.05)；A组未检测出NGF蛋白表达(OD值 0)，C组[OD值(0.343 2±0.006 81)]和D组[OD值(0.301 2±0.001 87)]较B组[OD值(0.2 83 7±0.007 69)] NGF蛋白表达明显增高，差异均有统计学意义(P<0.05)，E组[OD值(0.275 2±0.003 73)]与B组比较差异无统计学意义(P>0.05)；A组有少量NGF mRNA表达[OD值(0.286 6±0.004 41)]，C组[OD值(0.347 4±0.010 04)]和D组[OD值(0.329 4±0.019 22)]较B组[OD值0.301 2±0.001 87)] NGF mRNA表达明显增高，差异均有统计学意义(P<0.05)，E组[OD值(0.296 8±0.003 78)]与B组比较差异无统计学意义(P>0.05)。结论 CCK-8能够抑制缺氧缺血细胞模型凋亡，减轻神经细胞损伤的程度，其神经保护机制与促进NGF蛋白及NGF mRNA表达增加有关。
      【关键词】 胆囊收缩素；细胞凋亡；干预
      【中图分类号】 R-332 【文献标识码】 A 【文章编号】 1003—6350（2017）02—0180—03

Intervention effect of cholecystokinin-8 on hypoxic-ischemic cell damage model.
LIU Ying, ZHOU Jiang-bao.Department of Paediatrics, Panzhihua Maternal and Child Health-Care Hospital of Sichuan Province, Panzhihua 617000,Sichuan, CHINA
【Abstract】 Objective To investigate the intervention effect of cholecystokinin-8 (CCK-8) on hypoxic-isch-emic cell damage model, and to explore its effect on expression of nerve growth factor (NGF) protein and NGF mRNAin the neurons during apoptosis. Methods Neonatal rat cerebral cortical neurons cultured in vitro for 7 d were selectedas the blank control group (group A). Hypoxic-ischemic cell damage models (group B) were established by 50 mol/L glu-tamic acid soaking. The models treated with 1×10-6 mol/L, 1×10-7 mol/L, 1×10-8 mol/L were enrolled as group C, D, E,respectively, followed by continuous culture for 24 h. The apoptosis rate of the cells of five groups was detected by in si-tu end labeling technique, and the expression of NGF protein was detected by immunocytochemistry. The expression ofNGF mRNA was detected by in situ hybridization technique. Results The apoptosis rate of group A, group C andgroup D were respectively (8.49±4.54)%, (3.87±5.64)% and (7.84±4.19)%, which were significantly lower than (17.53±6.93)% in group B (P<0.05). The apoptosis rate of group E was (13.27±3.41)%, which showed no significant differencewith (17.53±6.93)% in group B (P>0.05). The expression of NGF protein was not detected in the group A (OD value: 0).NGF protein expression was significantly increased in group C (OD value: 0.343 2±0.006 81) and group D (OD value:0.301 2±0.001 87) than that in group B (OD value: 0.283 7±0.007 69), and the differences were statistically significant(P<0.05). There were no significant differences between group E (OD value: 0.275 2±0.003 73) and group B (P>0.05).There was a small amount of NGF mRNA expression in the group A (OD value: 0.286 6±0.004 41). NGF mRNA expres-sion were significantly increased in group C (OD value: 0.347 4±0.010 04) and group D (OD value: 0.329 4±0.019 22)than group B (OD value: 0.301 2±0.001 87), and the differences were statistically significant (P<0.05). There were nosignificant differences in NGF mRNA expression between group E (OD value: 0.296 8±0.003 78) and group B (P>0.05).Conclusion CCK-8 can suppress apoptosis of hypoxic-ischemic cell damage model, which neuroprotective mecha-nism was related to the increased expression of NGF protein and NGF mRNA.
      【Key words】 Cholecystokinin; Apoptosis; Intervention·论 著·doi:10.3969/j.issn.1003-6350.2017.02.003