摘要
目的探讨姜黄素(Cur)调节N-甲基-D-天冬氨酸受体(NMDAR)/钙离子(Ca^(2+))/钙调素依赖性蛋白激酶Ⅱ(CaMKⅡ)信号通路对异氟醚(ISO)诱导的幼龄小鼠术后认知功能障碍(POCD)的影响。方法将72只C57BL/6J小鼠分为对照组、ISO组、低剂量Cur组(Cur-L组,50 mg/kg)、中剂量Cur组(Cur-M组,100 mg/kg)、高剂量Cur组(Cur-H组,200 mg/kg)、Cur-H+NMDA(NMDAR激活剂)组(200 mg/kg+8 mg/kg),每组12只。经对应给药处理30 min后,对照组小鼠吸入含30%氧气和空气的混合气体2 h,其余各组小鼠吸入2%ISO 2 h,每天1次,持续14 d。末次给药24 h后,Morris水迷宫实验检测小鼠学习与空间记忆能力;HE染色检测海马CA1区病理学变化;免疫荧光染色检测小鼠海马CA1区神经元特异核蛋白(NeuN)阳性表达;TUNEL染色检测神经细胞凋亡;酶联免疫吸附试验检测海马CA1区组织中白细胞介素-1β(IL-1β)和肿瘤坏死因子-α(TNF-α)水平;蛋白印迹法检测海马CA1区组织中NMDAR1和CaMKⅡ蛋白表达;荧光探针检测海马CA1区Ca^(2+)浓度。结果与对照组比较,ISO组小鼠海马CA1区病理损伤严重,逃避潜伏期延长,神经细胞凋亡率升高,海马CA1区组织中IL-1β和TNF-α水平升高,NMDAR1和CaMKⅡ蛋白表达及Ca^(2+)浓度升高(P<0.05),穿越平台次数和NeuN阳性细胞数减少(P<0.05);与ISO组比较,Cur-L组、Cur-M组、Cur-H组小鼠海马CA1区病理损伤减轻,逃避潜伏期缩短,神经细胞凋亡率降低,海马CA1区组织中IL-1β和TNF-α水平降低,NMDAR1和CaMKⅡ蛋白表达及Ca^(2+)浓度降低(P<0.05),穿越平台次数和NeuN阳性细胞数增加(P<0.05),且呈剂量依赖性;NMDA减弱了高剂量Cur对ISO诱导的小鼠POCD的改善作用(P<0.05)。结论Cur可能通过抑制NMDAR/Ca^(2+)/CaMKⅡ信号通路改善ISO诱导的小鼠POCD。
Objective To investigate the effect of curcumin(Cur)regulating N-methyl-D-aspartate receptor(NMDAR)/calcium ion(Ca^(2+))/calmodulin dependent protein kinaseⅡ(CaMKⅡ)signaling pathway on isoflurane(ISO)-induced postoperative cognitive dysfunction(POCD)in young mice.Methods Seventy-two C57BL/6J mice were separated into the control group,the ISO group,the low-dose Cur group(Cur-L group,50 mg/kg),the medium-dose Cur group(Cur-M group,100 mg/kg),the high-dose Cur group(Cur-H group,200 mg/kg)and the Cur-H+NMDA(NMDAR activator)group(200 mg/kg+8 mg/kg),with 12 animals in each group.After 30 min of corresponding drug treatment,the mice in the control group inhaled a mixed gas containing 30%oxygen and air for 2 hours,and mice in the other groups inhaled 2%ISO for 2 hours,once a day for 14 days.Twenty-four hours after the last treatment,Morris water maze test was used to detect the learning and spatial memory abilities of mice.HE staining was used to detect the pathological changes in hippocampal CA1 area.Immunofluorescence staining was used to detect the positive expression of neuron specific nucleoprotein(NeuN)in hippocampus CA1 region of mice.TUNEL staining was used to detect neuronal apoptosis.Enzyme linked immunosorbent assay was used to detect levels of interleukin-1β(IL-1β)and tumor necrosis factor-α(TNF-α)in hippocampal CA1 tissue.Western blot assay was used to detect NMDAR1 and CaMKⅡprotein expression in mouse hippocampal CA1 tissue.Intracellular Ca^(2+)concentration in hippocampal CA1 region was detected by fluorescence probe.Results Compared with the control group,the pathological damage of hippocampal CA1 region of mice was severe in the ISO group,and the escape latency was prolonged.The apoptosis rate of neural cells,levels of IL-1βand TNF-α,expression levels of NMDAR1 and CaMKⅡprotein,and the concentration of Ca^(2+)in hippocampal CA1 region were increased(P<0.05).Times of crossing platform and the number of NeuN positive cells were decreased in the ISO group(P<0.05).Compared with the ISO group,pathological damages of hippocampal CA1 region of mice were alleviated in the Cur-L group,the Cur-M group and the Cur-H group,the escape latency was shortened,and the apoptosis rate of neural cells,levels of IL-1βand TNF-α,expression levels of NMDAR1 and CaMKⅡprotein,and the concentration of Ca^(2+)in hippocampal CA1 region were decreased(P<0.05).Times of crossing platform and the number of NeuN positive cells were increased(P<0.05),which was dosedependent.NMDA attenuated the improvement effect of high-dose Cur on ISO induced POCD in mice(P<0.05).Conclusion Curcumin may improve ISO-induced POCD in mice by inhibiting NMDAR/Ca^(2+)/CaMKⅡsignaling pathway.
作者
马慧敏
柳璐
熊英
赵慧
孔繁丽
MA Huimin;LIU Lu;XIONG Ying;ZHAO Hui;KONG Fanli(Anesthesia Department Operating Room,Wuhan Children's Hospital Affiliated to Tongji Medical College of Huazhong University of Science and Technology(Wuhan Maternal and Child Health Hospital),Wuhan 430014,China)
出处
《天津医药》
CAS
北大核心
2023年第9期948-954,共7页
Tianjin Medical Journal
基金
湖北省卫生健康委科研立项项目(WJ2019M012)。
作者简介
马慧敏(1983),女,主治医师,主要从事麻醉学方面研究。E-mail:mahuimin972@163.com;通信作者:柳璐,E-mail:365038103@qq.com。
