三白草酮通过调控海马Nrf2/HO-1表达减少酒精戒断大鼠抑郁样行为  

Sauchinone attenuates alcohol withdrawal-induced depression-like behaviors in rats by modulating hippocampal Nrf2/HO-1 expression

作　　者：姜敏 李露露 吴桐[2] 李成冲[2] 王宇晨 焦宇 刘艺涵 马淑丽 郑晓宇 赵容杰 赵正林 JIANG Min;LI Lu-lu;WU Tong;LI Cheng-chong;WANG Yu-chen;JIAO Yu;LIU Yi-han;MA Shu-li;ZHENG Xiao-yu;ZHAO Rong-jie;ZHAO Zheng-lin(School of Medical Technology,Qiqihar Medical University,Qiqihar 161006,China;School of Mental Health,Qiqihar Medical University,Qiqihar 161006,China)

机构地区：[1]齐齐哈尔医学院医学技术学院,黑龙江齐齐哈尔161006 [2]齐齐哈尔医学院精神卫生学院,黑龙江齐齐哈尔161006

出　　处：《中国病理生理杂志》2022年第10期1803-1811,共9页Chinese Journal of Pathophysiology

基　　金：齐齐哈尔市科技计划联合引导项目(No.LHYD-202011);齐齐哈尔市科技计划联合引导项目(No.LHYD-2021015)。

摘　　要：目的:探讨三白草酮(sauchinone,Sau)对酒精戒断大鼠抑郁样行为可能的影响及其机制。方法:取成年雄性Sprague-Dawley大鼠,随机分为正常对照组、酒精戒断模型组、低剂量Sau治疗组和高剂量Sau治疗组,每组8只。正常对照组腹腔注射等体积生理盐水,其余3组每日1次腹腔注射酒精3 g/kg,连续28 d,然后戒断3 d诱导酒精戒断模型。在戒断期间,Sau低、高剂量治疗组分别每日1次腹腔注射5和25 mg/kg Sau,连续3 d。末次给药30 min后通过蔗糖偏好实验、悬尾实验(tail suspension test,TST)和强迫游泳实验(forced swimming test,FST)检测大鼠行为学变化;采用商品化试剂盒测定海马组织中总超氧化物歧化酶(total superoxide dismutase,T-SOD)、过氧化氢酶(catalase,CAT)、丙二醛(malondialdehyde,MDA)和谷胱甘肽(glutathione,GSH)水平;Western blot检测海马核因子E2相关因子2(nuclear factor E2-related factor 2,Nrf2)和血红素加氧酶1(heme oxygenase-1,HO-1)蛋白表达。使用H2O2(200μmol/L)诱导小鼠海马神经元HT22细胞氧化应激模型后,分别给予低、高剂量(3和30μmol/L)Sau干预,通过MTT法测定细胞活力;流式细胞术检测细胞凋亡水平;采用试剂盒测定细胞内T-SOD活性和MDA含量;免疫荧光法检测细胞核和细胞质Nrf2蛋白的表达。结果:与酒精戒断模型组比较,低、高剂量Sau治疗组大鼠蔗糖偏好率显著升高(P<0.05或P<0.01),TST和FST的不动时间均显著降低(P<0.05或P<0.01),大鼠海马中T-SOD、CAT和GSH水平显著升高而MDA含量显著降低(P<0.05或P<0.01),同时大鼠海马中Nrf2和HO-1蛋白表达显著升高(P<0.05或P<0.01)。体外实验显示,与氧化应激模型组比较,低、高剂量Sau组HT22细胞活力显著升高(P<0.05或P<0.01),细胞凋亡水平显著降低(P<0.05或P<0.01),细胞内T-SOD活性显著升高而MDA含量显著降低(P<0.05或P<0.01),同时高剂量Sau组细胞核Nrf2蛋白表达显著降低(P<0.01)。结论:Sau减少酒精戒断大鼠的抑郁样行�AIM:To investigate the effect of sauchinone(Sau)on depression-like behaviors in alcohol withdrawal rats and its relevant mechanism.METHODS:Adult male Sprague-Dawley rats were randomly divided into control group,alcohol withdrawal model group,low-dose Sau treatment group and high-dose Sau treatment group,with 8 rats in each group.The rats in control group were intraperitoneally injected with saline,while those in the other 3 groups were intraperitoneally injected with alcohol(3 g/kg)once a day for 28 consecutive days followed by 3 abstinent days to establish alcohol withdrawal model.During the withdrawal period,the rats in low-and high-dose Sau treatment groups were intraperitoneally injected with 5 and 25 mg/kg of Sau once a day for 3 consecutive days.Thirty minutes after the injection of Sau,the behavioral changes of the rats were detected by sucrose preference test,tail suspension test(TST)and forced swimming test(FST).The levels of total superoxide dismutase(T-SOD),malondialdehyde(MDA),catalase(CAT)and glutathione(GSH)in hippocampus were determined by the commercial available kits.The protein levels of hippocampal nuclear factor E2-related factor 2(Nrf2)and heme oxygenase-1(HO-1)were detected by Western blot.Additionally,the oxidative stress model of mouse hippocampal neuronal HT22 cells was induced by H2O2(200μmol/L),and the cells were treated with low-dose(3μmol/L)or high-dose(30μmol/L)Sau.The cell viability was determined by MTT assay.The cell apoptosis was detected by flow cytometry.The activity of T-SOD and the content of MDA in cells were determined by the kits.The levels of Nrf2 protein in the nucleus and cytoplasm were measured by immunofluorescence.RESULTS:The behavioral tests showed that compared with alcohol withdrawal model group,the sucrose preference rates of the rats in lowdose and high-dose Sau treatment groups were significantly increased(P<0.05 or P<0.01),and the immobility time of TST and FST was significantly decreased(P<0.05 or P<0.01).The levels of T-SOD,CAT and GSH in the hippocampus

关 键 词：三白草酮 酒精戒断 抑郁症 氧化应激 核因子E2相关因子2 血红素加氧酶1 

分 类 号：R749.62[医药卫生—神经病学与精神病学] R363.2[医药卫生—临床医学]