三七茎叶总皂苷抑制七氟醚诱导的海马神经元凋亡机制研究  

Mechanism of Stem-Leaf Saponins from Panax Notoginseng Inhibiting Sevoflurane-Induced Neuronal Apoptosis

作　　者：文艳 温旭 李靖[3] 李洋 黄先全 刘康 WEN Yan;WEN Xu;LI Jing;LI Yang;HUANG Xianquan;LIU Kang(Nanchong Traditional Chinese Medicine Hospital,Nanchong,Sichuan,China 637000;Mianyang 404 Hospital,Mianyang,Sichuan,China 621000;People′s Hospital of Yilong County,Nanchong,Sichuan,China 637600;Nanchong Central Hospital,Nanchong,Sichuan,China 637000)

机构地区：[1]四川省南充市中医医院,四川南充637000 [2]四川绵阳四○四医院,四川绵阳621000 [3]四川省仪陇县人民医院,四川南充637600 [4]四川省南充市中心医院,四川南充637000

出　　处：《中国药业》2023年第21期61-67,共7页China Pharmaceuticals

摘　　要：目的探讨三七茎叶总皂苷(PNSLS)调节七氟醚诱导海马神经元凋亡的作用机制。方法以不同剂量七氟醚(0,4.1%,8%)干预小鼠海马神经元细胞系HT22,使用不同质量浓度的PNSLS(1.5,3.0,6.0,12.0μg/mL)预处理神经元。将HT22细胞分为对照组、暴露于4.1%七氟醚的细胞组(4.1%Sev组)、使用6μg/mL PNSLS对细胞进行预处理1 h后将细胞暴露于4.1%七氟醚组(4.1%Sev+PNSLS组)、在PNSLS处理前1 h加入丝裂原激活蛋白激酶(MAPK)激活剂anisomycin(An)再暴露于七氟醚和PNSLS的共培养组(4.1%Sev+PNSLS+An组)。通过CCK-8法检测神经元活力,流式细胞术及免疫印迹(Western blot)法检测海马神经元凋亡,比色活性测定试剂盒测定胱天蛋白酶-3(caspase-3)活性,Western blot法测定内质网应激相关蛋白及p38 MAPK/核转录因子-κB p65(p38 MAPK/NF-κB p65)信号通路相关蛋白的表达水平。结果4.1%七氟醚显著抑制海马神经元活力(P<0.05),6μg/mL PNSLS显著升高神经元活力并抑制海马神经元凋亡(P<0.05)。与Sev组比较,Sev+PNSLS组细胞中凋亡蛋白Bax表达水平显著降低(P<0.05),B细胞淋巴瘤2家族蛋白(Bcl-2)表达水平显著升高(P<0.05),caspase-3活性显著降低(P<0.05);Sev+PNSLS组内质网应激诱导的凋亡信号通路相关蛋白78000葡糖调节蛋白(GRP78)、内质网应激相关蛋白(CHOP)表达水平均显著降低(P<0.01),p38和IκBα的磷酸化水平均显著降低(P<0.01),细胞质中NF-κB p65水平显著升高(P<0.01),细胞核中NF-κB p65水平显著降低(P<0.01)。与Sev+PNSLS组比较,Sev+PNSLS+An组p38和IκBα的磷酸化水平均显著升高(P<0.01),细胞质中NF-κB p65水平显著降低(P<0.01),细胞核中NF-κB p65水平显著升高(P<0.01),海马神经元活力显著降低(P<0.01),凋亡率显著升高(P<0.01)。结论PNSLS通过激活p38 MAPK/NF-κB p65信号通路抑制七氟醚诱导的神经元凋亡。Objective To investigate the mechanism of stem-leaf saponins from Panax notoginseng(PNSLS)regulating the apoptosis of sevoflurane(Sev)-induced neuronal apoptosis.Methods Different doses of sevoflurane(0,4.1%,8%)were used to intervene in the mouse hippocampal neuronal cell line HT22,and different concentrations of PNSLS(1.5,3.0,6.0,12.0μg/mL)were used to pre-treat the neurons.HT22 cells were divided into the control group,4.1%Sev group(cells were exposed to 4.1%Sev),and 4.1%Sev+PNSLS group(cells were pre-treated with 6μg/mL PNSLS for 1 h,and then they were exposed to 4.1%Sev),and 4.1%Sev+PNSLS+An group[MAPK activator anisomycin(An)was added 1 h before PNSLS treatment,and then the cells were co-cultured with Sev and PNSLS].Neuronal activity was detected by the CCK-8 assay,the hippocampal neuronal apoptosis was detected by flow cytometry and Western blot,the activity of caspase-3 was determined by the colorimetric activity assay kit,and the expression levels of endoplasmic reticulum stress-related proteins and p38 mitogen-activated protein kinase/nuclear transcription factor-κB p65(p38 MAPK/NF-κB p65)signaling pathway-related proteins were determined by Western blot.Results 4.1%Sev could significantly inhibit hippocampal neuronal activity(P<0.05),6μg/mL PNSLS could significantly increase neuronal activity and inhibit hippocampal neuronal apoptosis(P<0.05).Compared with those in the Sev group,the expression level of PNSLS apoptotic protein Bax showed a significant decrease(P<0.05),the expression level of Bcl-2 showed a significant increase(P<0.05),and the caspase-3 activity showed a significant decrease in the Sev+PNSLS group(P<0.05);the expression levels of apoptosis signaling pathway-related proteins GRP78 and CHOP induced by endoplasmic reticulum stress in the Sev+PNSLS group significantly reduced(P<0.01);the phosphorylation levels of p38 and IκBαsignificantly reduced(P<0.01),the levels of NF-κB p65 in the cytoplasm significantly increased(P<0.01),and the levels of NF-κB p65 in the nucleus significantly r

关 键 词：七氟醚 三七茎叶总皂苷 P38丝裂原激活蛋白激酶 核转录因子-κB p65 胱天蛋白酶-3 神经元凋亡 内质网应激 

分 类 号：R932[医药卫生—生药学] R285.5[医药卫生—药学]