摘要
目的:基于脂质组学研究越鞠丸对抑郁小鼠血清、前额叶皮层、海马区脂质代谢的影响,探讨其改善脂质代谢防治抑郁的可能途径.方法:7周龄的C57BL/6小鼠随机分为空白组、模型组、越鞠丸组(3.6 g·kg-1)和氟西汀组(10mg·kg-1),采用慢性不可预知刺激诱导抑郁模型,造模3周后,各给药组按剂量灌胃相应药液,空白组和模型组小鼠给予等体积去离子水灌胃,1次/d,持续2周.给药结束后,通过糖水偏好实验、旷场实验、悬尾实验、强迫游泳实验等神经行为学指标评价越鞠丸的抗抑郁作用;生化检测小鼠血清中总胆固醇(TC)、甘油三酯(TG)、低密度脂蛋白胆固醇(LDL-C)、高密度脂蛋白胆固醇(HDL-C)、天冬氨酸氨基转移酶(AST)和丙氨酸氨基转移酶(ALT)含量;基于超高效液相色谱-线性离子阱-静电场轨道阱质谱法(UPLC-LTQ-OrbitrapMS)对小鼠血清、前额叶皮层、海马区进行脂质组学分析;蛋白免疫印迹法(Western blot)检测小鼠胃组织中哺乳动物雷帕霉素靶蛋白(mTOR)、核糖体蛋白S6激酶(S6K)、磷酸化(p)-mTOR、p-S6K蛋白的表达情况.结果:与空白组比较,模型组小鼠糖水偏好率和旷场中心运动时间显著减少(P＜0.01),悬尾不动时间和强迫游泳不动时间显著增加(P＜0.01),血清TC、TG、LDL-C、HDL-C、AST和ALT水平明显升高(P＜0.05,P＜0.01);与模型组比较,越鞠丸组小鼠的糖水偏好率和旷场中心运动时间显著增加(P＜0.01),悬尾不动时间和强迫游泳不动时间显著减少(P＜0.01),血清TC、TG、LDL-C、AST和ALT水平明显降低(P＜0.05,P＜0.01).脂质组学分析显示,越鞠丸对抑郁小鼠的血清、前额叶皮层、海马区的脂质代谢均有明显影响,差异脂质代谢物主要富集于甘油磷脂代谢、鞘脂信号和糖基磷脂酰肌醇-锚定蛋白生物合成等通路上,其中又以甘油磷脂的代谢通路最为显著.Western blot结果显示,与空白组比较,模型组小鼠胃组织中p-mTOR/mTOR和p-S6K/S6K的表达水平显著升高(P＜0.01);与模型组比较,越鞠丸组小鼠胃组织中p-mTOR/mTOR和p-S6K/S6K的表达水平显著降低(P＜0.01).结论:越鞠丸对脂质代谢的干预是其发挥抗抑郁效应的潜在途径之一,可能与胃中脂质代谢上游mTOR/S6K信号通路的调控相关.
Abstract
Objective:To investigate the effect of Yuejuwan on lipid metabolism in serum,prefrontal cortex and hippocampus of depressed mice based on lipidomics,and to explore the potential pathways for improving lipid metabolism to prevent depression.Methods:Seven-week-old C57BL/6 mice were randomly divided into blank group,model group,Yuejuwan group(3.6 g·kg-1)and fluoxetine group(10 mg·kg-1),and chronic unpredictable mild stress(CUMS)was used to establish the depression model.After 3 weeks of modeling,each administration group was gavaged with the corresponding drug solution according to the dose,and mice in the blank and model groups were given an equal volume of deionised water by gavage,one time/d for 2 weeks.After administration,the antidepressant effect of Yuejuwan was evaluated by neurobehavioral indices such as sucrose preference test,open field test,tail suspension test and forced swimming test.An automatic biochemical analyzer was used to measure contents of total cholesterol(TC),triglyceride(TG),low-density lipoprotein cholesterol(LDL-C),high-density lipoprotein cholesterol(HDL-C),aspartate aminotransferase(AST)and alanine aminotransferase(A LT)in mouse serum.Lipidomic analysis of mouse serum,prefrontal cortex and hippocampus was performed based on ultra-performance liquid chromatography-linear ion trap-electrostatic field orbitrap mass spectrometry(UPLC-LTQ-Orbitrap-MS),and the expression of mammalian target of rapamycin(mTOR),ribosomal protein S6 kinase(S6K),phosphorylation(p)-mTOR,p-S6K in gastric tissues of mice was detected by Western blot.Results:Compared with the blank group,mice in the model group exhibited significantly reduced sucrose preference rate and center movement time in the open field test(P＜0.01),the immobility times in the tail suspension test and forced swimming test were significantly increased(P＜0.01),and serum levels of TC,TG,LDL-C,HDL-C,AST and ALT were significantly elevated(P＜0.05,P＜0.01).Compared with the model group,the Yuejuwan group showed a significant increase in the sucrose preference rate and center movement time in the open field test(P＜0.01),the immobility times in the tail suspension test and forced swimming test were significantly reduced(P＜0.01),and the serum levels of TC,TG,LDL-C,AST and ALT were significantly decreased(P＜0.05,P＜0.01).Lipidomic analysis revealed that Yuejuwan had a significant effect on lipid metabolism in serum,prefrontal cortex and hippocampus of depressed mice,and The differential lipid metabolites were mainly enriched in the metabolic pathways of glycerophospholipid metabolism,sphingolipid signaling,and glycosylphosphatidylinositol-anchored protein biosynthesis,among which the glycerophospholipid metabolic pathway was the most significant.Western blot results showed that compared with the blank group,the relative expression levels of p-mTOR/mTOR and p-S6K/S6K in the gastric tissues of mice in the model group were significantly increased(P＜0.01).In comparison with the model group,the relative expression levels of p-mTOR/mTOR and p-S6K/S6K in the gastric tissues of mice in the Yuejuwan group were significantly decreased(P＜0.01).Conclusion:The intervention of Yuejuwan on lipid metabolism is one of the potential pathways for its antidepressant effect,which may be related to the regulation of mTOR/S6K signaling pathway upstream of lipid metabolism in the gastric tissues.
国家科技期刊平台
NSTL
维普
万方数据