栝楼桂枝汤调控沉默信息调节因子1通路介导的氧化应激对神经细胞损伤的保护作用  

Protective effect of Gualou Guizhi decoction on neuron injury via silent information regulator 1 mediated oxidative stress

作　　者：胡海霞[1] 杨劲博 林心君[2] 钟兴华 HU Hai-xia;YANG Jin-bo;LIN Xin-jun;ZHONG Xing-hua(Innovation and Transformation Center,Fujian University of Traditional Chinese Medicine,Fuzhou 350122,Fujian Province,China;Institute of Integrated Chinese and Western Medicine,Fujian University of Traditional Chinese Medicine,Fuzhou 350122,Fujian Province,China;Academy of Integrative Medicine,Fujian University of Traditional Chinese Medicine,Fuzhou 350122,Fujian Province,China)

机构地区：[1]福建中医药大学,科技创新与转化中心,福建福州350122 [2]福建中医药大学,中西医结合学院,福建福州350122 [3]福建中医药大学,中西医结合研究院,福建福州350122

出　　处：《中国临床药理学杂志》2023年第12期1738-1742,共5页The Chinese Journal of Clinical Pharmacology

基　　金：福建省自然科学基金资助项目(2021J01937)。

摘　　要：目的探讨栝楼桂枝汤(GLGZD)调控沉默信息调节因子1(Sirt1)信号通路介导的氧化应激发挥保护神经细胞的作用机制。方法将HT22细胞分为对照组、模型组和低、中、高剂量实验组。对照组为常规培养;模型组缺氧培养4 h并复氧24 h制备氧糖剥夺再灌注(OGD/R)模型;低、中、高剂量实验组在模型建立后,分别加入200、400和800μg·mL^(-1)GLGZD继续培养24 h。用细胞计数试剂盒检测细胞生长活力,用试剂盒检测细胞上清液中乳酸脱氢酶(LDH)、丙二醛(MDA)和谷胱甘肽过氧化物酶(GSH-PX)含量,用激光共聚焦显微镜观察活性氧(ROS)的表达情况,用流式细胞仪检测细胞凋亡率,用蛋白质印迹法检测Sirt1、过氧化物酶体增殖活化受体γ辅助活化因子1α(PGC-1α)和雌激素相关受体α(ERRα)的表达水平。结果中剂量实验组、模型组和对照组的LDH含量分别为(217.17±6.64)、(336.53±13.05)和(268.22±5.89)U·L^(-1),MDA含量分别为(2.57±0.16)、(4.38±0.41)和(2.74±0.30)nmol·mL^(-1),GSH-PX含量分别为(111.21±4.27)、(95.83±4.12)和(115.90±5.82)mol·L^(-1),ROS荧光强度分别为19.91±4.39、67.33±5.91和9.20±1.13,细胞凋亡率分别为(11.87±2.33)%、(22.96±0.91)%和(8.75±0.28)%,Sirt1蛋白相对表达水平分别为1.02±0.32、0.38±0.29和1.00±0.13,PGC-1α蛋白相对表达水平分别为1.02±0.58、0.33±0.07和1.02±0.24,ERRα蛋白相对表达水平分别为1.28±0.19、0.50±0.13和1.00±0.11。中剂量实验组的上述指标与模型组比较,差异均有统计学意义(P<0.05,P<0.01)。结论GLGZD可能通过调控Sirt1信号通路抑制脑缺血后的氧化应激水平和神经细胞凋亡,进而改善神经组织损伤。Objective To investigate the neuroprotective effect of Gualou Guizhi decoction(GLGZD)on oxidative stress through silent information regulator 1(Sirt1)signaling pathway.Methods HT22 cells were divided into control group,model group and experimental-L,-M,-H groups.Control group was cultured normally;model group was exposed to 4 h hypoxia followed by 24 h reoxygenation;experimental-L,-M,-H groups were treated with 200,400 and 800μg·mL^(-1)GLGZD for 24 h after establishing cell model.Cell viability was measured by cell counting kit assay.The contents of lactate dehydrogenase(LDH),malonic dialdehyde(MDA)and glutathione peroxidase(GSH-PXw)ere detected in cell culture supernatant.Reactive oxygen species(ROS)was observed by laser scanning confocal microscopy.Apoptosis rate was examined by flow cytometry.The expression levels of Sirtl,Peroxisome proliferator-activated receptor-co-activator-lα(PCC-lα),estrogen-related receptorα(ERRα)were detected by Western blotting.ResultsLIDH contents of experimental-M,model and control groups were(217.17±6.64),(336.53±13.05)and(268.22±5.89)U·L^(-1);MDA contents were(2.57±0.16),(4.38±0.41)and(2.74±0.30)nmol·mL^(-1);GSH-PX contents were(111.21±4.27),(95.83±4.12)and(115.90±5.82)mol·L^(-1);ROS fluorescence intensities were 19.91±4.39,67.33±5.91 and 9.20±1.13;apoptosis rates were(11.87±2.33)%,(22.96±0.91)%and(8.75±0.28)%;the relative expression levels of Sirtl protein were 1.02±0.32,0.38±0.29 and 1.00±0.13;the relative expression levels of PGC-1αprotein were 1.02±0.58,0.33±0.07 and 1.02±0.24;the relative expression levels of ERRαprotein were 1.28±0.19,0.50±0.13 and 1.00±0.11,respectively.There were statistically significant differences in the above indexes between experimental-M group and model group(P<0.05,P<0.01).Conclusion CLGZD may improve neural tissue damage via regulating Sirtl signaling pathway to inhibit oxidative stress and the neuronal apoptosis after cerebral ischemia.

关 键 词：栝楼桂枝汤 缺血性脑卒中 沉默信息调节因子1信号通路 氧化应激 凋亡 

分 类 号：R28[医药卫生—中药学]