摘要
目的探讨纳洛酮对七氟醚诱导的胎鼠神经干细胞(NSCs)凋亡的影响。方法将NSCs分为空白组、对照组和低、中、高剂量实验组。空白组给予正常培养;对照组通入含3%七氟醚的混合气体培养48 h;低、中、高剂量实验组均通入含3%七氟醚的混合气体,并分别加入含0.1,1.0和10.0μmol·L^(-1)纳洛酮溶液的DMEM-12F培养基共培养48 h。用流式细胞仪检测NSCs的凋亡情况,用蛋白质印迹法检测沉默信息调节因子1(SIRT1)、磷脂肌醇3-激酶(PI3K)、蛋白激酶B(AKT)和哺乳动物西罗莫司靶蛋白(mTOR)蛋白的表达水平。结果低、高剂量实验组和对照组、空白组的NSCs凋亡率分别为(32.25±4.83)%,(16.77±2.52)%,(43.59±6.53)%和(8.66±1.29)%;SIRT1蛋白表达水平分别为0.50±0.08,0.99±0.15,0.20±0.03和1.27±0.19;p-PI3K/PI3K蛋白相对表达水平分别为0.85±0.13,0.47±0.04,1.06±0.16和0.25±0.03;p-AKT/AKT蛋白相对表达水平分别为0.90±0.14,0.50±0.06,1.13±0.17和0.32±0.05;p-mTOR/mTOR蛋白相对表达水平分别为0.99±0.15,0.55±0.08,1.26±0.19和0.35±0.05。低、高剂量实验组的上述指标与对照组比较,差异均有统计学意义(均P<0.05)。结论纳洛酮能有效抑制七氟醚诱导的NSCs凋亡,其机制可能与上调SIRT1表达,进而抑制PI3K/AKT/mTOR信号通路激活有关。
Objective To investigate the effect of naloxone on the apoptosis of sevoflurane-induced neural stem cells(NSCs)in fetal rats.Methods NSCs were divided into blank group,control group and experimental-L,-M,-H groups.The blank group was given normal culture;the control group was given mixed gas containing 3%sevoflurane for 48 h;the experimental-L,-M,-H groups were given mixed gas containing 3%sevoflurane and co-cultured with DMEM-12F medium containing 0.1,1.0 and 10.0μmol·L^(-1) naloxone solution for 48 h,respectively.Flow cytometry was used to detect the apoptosis of NSCs.Western blot was used to detect the expression levels of silent information regulator 1(SIRT1),phosphatidylinositol 3-kinase(PI3K),protein kinase B(AKT)and mammalian target of rapamycin(mTOR)protein.Results The apoptosis rates of experimental-L,-H groups and control group,blank group were(32.25±4.83)%,(16.77±2.52)%,(43.59±6.53)%and(8.66±1.29)%;the expression levels of SIRT1 protein were 0.50±0.08,0.99±0.15,0.20±0.03 and 1.27±0.19;the relative expression levels of p-PI3K/PI3K were 0.85±0.13,0.47±0.04,1.06±0.16 and 0.25±0.03;the relative expression levels of p-AKT/AKT were 0.90±0.14,0.50±0.06,1.13±0.17 and 0.32±0.05;the relative expression levels of p-mTOR/mTOR protein were 0.99±0.15,0.55±0.08,1.26±0.19 and 0.35±0.05.The differences were statistically significant between experimental-L,-H groups and control group(all P<0.05).Conclusion Naloxone can effectively inhibit the apoptosis of sevoflurane-induced NSCs in fetal rats,and its mechanism may be related to up-regulate SIRT1 expression and inhibite the activation of PI3K/AKT/mTOR signaling pathway.
作者
李幸雷
虎琳
李军仕
苏孟勤
LI Xing-lei;HU Lin;LI Jun-shi;SU Meng-qin(Department of Anesthesiology,Henan Provincial Hospital,Zhengzhou 450000,Henan Province,China;Department of Anesthesiology,Hainan Third People’s Hospital,Haikou 570000,Hainan Province,China)
出处
《中国临床药理学杂志》
CAS
CSCD
北大核心
2022年第8期822-826,共5页
The Chinese Journal of Clinical Pharmacology
基金
河南省医学科技计划联合共建基金资助项目(LHGJ20190840)。
