摘要
目的探讨乌司他丁(UTI)通过调控核苷酸结合寡聚化结构域样受体蛋白3(NLRP3)表达影响神经元凋亡,发挥保护受损神经元的作用和分子机制。方法培养原代神经元至成熟,随机分为空白组(Sham组)、对照组(Control组)和乌司他丁预处理组(UTI组)。先进行UTI组和对照组预处理72 h,再进行氧糖剥夺损伤3 h、6 h,最后恢复正常培养液培养2 h后取材。利用荧光免疫组化法检测NLRP3的空间表达情况;利用蛋白质印迹法(Western-blot)检测NLRP3的蛋白水平表达变化。利用细胞计数试剂盒(cell counting kit-8,CCK-8)检测试剂盒检测神经元凋亡情况。结果与对照组比较,氧糖剥夺6 h后UTI组的NLRP3蛋白表达明显降低(P<0.05),且神经元凋亡数量显著减少,各组间差异有统计学意义(P<0.05);而氧糖剥夺3 h后UTI组的NLRP3蛋白表达和神经元凋亡数量无明显变化(P>0.05)。结论 UTI预处理调控NLRP3的表达,可能是减轻神经元凋亡的重要原因之一,具有显著的保护受损神经细胞的作用。
Objective The potential therapeutic effect and the related mechanism of ulinastatin（ UTI）and nucleotide-binding oligomerization domain receptor protein 3（ NLRP3） on cultured neuron associated with hypoxic-ischemic damage were investigated. Methods Neurons were in vitro cultivated. The cells were randomly divided into the sham group（ Sham group）,physiological saline control group（ Control group）,the damage group（ OGD group） and the UTI pretreatment group（ UTI group）. Control group and UTI group were performed the pretreatment for 72 h,and then all the groups were subjected to oxygen-glucose deprivation（ OGD） for 3 h and 6 h,lastly,all the neurons were cultivated by normal culture for another 2 h and then sampled. The expression of NLRP3 was detected by immunofluorescence staining; the expression of NLRP3 was detected by Western blotting. Neurons viability was detected by cell counting kit-8（ CCK-8）.Results The expression of NLRP3 protein was different among Sham group,Control group,and UTI group（P〈0. 05）; after OGD for 6 h,UTI could obviously reduce the expression of NLRP3 protein and the number of apoptotic neurons compared with Control group（ P〈0. 05）. But after OGD for 3 h,the expression of NLRP3 protein and the number of apoptotic neurons did no change significantly in UTI group compared with Control group（ P〈0. 05）. Conclusion The UTI pretreatment can effectively enhance the brain damage nerve function which maybe through regulating NLRP3 expression following the ischemic brain injury.
作者
李笑冰
岳宗源
白婧
任垒
饶维
彭程
费舟
张磊
LI Xiaobing;YUE Zongyuan;BAI Jing;REN Lei;RAO Wei;PENG Cheng;FEI Zhou;ZHANG Lei(Brigade of Cadets, Air Foree Military Medical University, Xi＇an 710032;Unit 91709 of People＇s Liberation Army, Huzhun 133300;Department of Neurosurgery, Xijing Hospital, Air Force Military Medical University, Xi＇an 710032;Department of Neurosurgery, No. 261st Hospital of People＇s Liberation Army, Beijing 100094, China)
出处
《中华神经外科疾病研究杂志》
CAS
2018年第2期147-150,共4页
Chinese Journal of Neurosurgical Disease Research
基金
国家自然科学基金资助项目(81200949
81301098)
全军医学科技青年培育计划孵化项目资助项目(16QNP113)
关键词
NOD样受体蛋白3
乌司他丁
缺氧缺血性损伤
神经元
NOD-like receptor family pyrin domain containing 3
Ulinastatin
Hypoxic-ischemicdamage
Neuron
