摘要
目的:探讨苦参子提取物的抗抑郁作用及其分子作用机制。方法:采用腹腔注射脂多糖(LPS)建立小鼠抑郁模型,设置正常组、模型组、氟西汀组(2.5 mg·kg^(-1)),以及苦参子低、中、高剂量组(200、400、800 mg·kg^(-1)),连续灌胃7 d,利用旷场实验、高架十字迷宫实验和强迫游泳实验考察苦参子提取物的抗抑郁作用。苏木素-伊红(HE)染色观察小鼠海马组织病理形态变化,蛋白免疫印迹法(Western blot)检测小鼠脑组织中G_(1)/S特异性周期蛋白D_(1)(Cyclin D_(1))、Wnt1蛋白、β-连环蛋白(β-catenin)、磷酸化糖原合成酶激酶-3β(p-GSK-3β)的蛋白表达水平。通过原位末端标记法(TUNEL)检测海马细胞凋亡情况。结果:小鼠行为学结果显示,与正常组比较,模型组的旷场运动速度、旷场中间区运动距离、高架十字迷宫开臂停留时间均显著降低(P<0.01),强迫游泳实验不动时间明显增加(P<0.05)。与模型组比较,苦参子中、高剂量组的旷场运动速度、开臂停留时间增加(P<0.05,P<0.01),强迫游泳实验不动时间降低(P<0.05);苦参子高剂量组小鼠旷场中间区运动距离增加(P<0.05)。HE染色结果发现与正常组比较,模型组小鼠海马神经元结构损伤。与模型组比较,给予苦参子低、中、高剂量组干预后,小鼠海马神经元结构病理状态减轻,神经元增多、排列整齐、胞浆清楚。Western blot结果显示,小鼠注射LPS后脑组织中Wnt1及β-catenin蛋白表达水平显著降低(P<0.01),Cyclin D_(1)及p-GSK-3β蛋白表达水平显著增加(P<0.01);与模型组比较,苦参子中、高剂量组小鼠脑组织中Wnt1和β-catenin蛋白表达水平均显著增加(P<0.01),而Cyclin D_(1)和p-GSK-3β蛋白表达量则显著降低(P<0.01)。TUNEL染色结果显示,模型组小鼠的海马细胞凋亡率较正常组显著增加(P<0.01),与模型组比较,苦参子中、高剂量组小鼠海马细胞凋亡率显著降低(P<0.01)。结论:苦参子提取物可有效改善LPS所致抑郁小鼠的抑郁严重程度,其分子机制与调控Wnt/β-catenin信号转导通路介导的神经炎症和海马神经元凋亡有关。
Objective:To explore the antidepressant effect of Sophora flavescens seed extract and its molecular mechanism.Method:A mouse depression model was established by intraperitoneal injection of lipopolysaccharide(LPS),and normal group,model group,fluoxetine group(2.5 mg·kg^(-1)),and S.flavescens seed low,medium and high dose groups(200,400,800 mg·kg^(-1))were set up for 7 d of consecutive gavage.Then the antidepressant effect of S.flavescens seed extract was evaluated by using open field test,elevated plus maze test and forced swimming test.Pathological morphological changes in the hippocampal tissue was observed by hematoxylin-eosin(HE)staining.Protein expression levels of G_(1)/S-specific cyclin D_(1)(Cyclin D_(1)),Wnt1,β-catenin and phosphorylated glycogen synthase kinase-3β(p-GSK-3β)in mouse brain tissues were detected by Western blot.Hippocampal cell apoptosis was detected by terminal deoxynucleotidyl transferase mediated deoxyuridine triphosphate(dUTP)nick end labeling(TUNEL).Result:The results of mouse behavioral experiments showed that compared with the normal group,the speed of movement in the open field and the distance of movement in the central area of the open field,and the time spent on the open arms of the elevated plus maze were significantly reduced in the model group(P<0.01),while immobility time in the forced swimming test was significantly increased(P<0.05).Compared with the model group,the S.flavescens seed medium and high dose groups had increased speed of movement in the open field test and time spent on the open arms of the elevated plus maze test(P<0.05,P<0.01),and decreased immobility time in the forced swimming test(P<0.05),the distance of movement in the central area of the open field test increased in the high dose group(P<0.05).HE staining results showed that compared with the normal group,the hippocampal neuron structure of mice in the model group was damaged.Compared with the model group,after treatment of S.flavescens seed extract,the pathological state of the mouse hippocampal neuron structure was alleviated,and the neurons increased,were neatly arranged,and the cytoplasm was clear.Western blot results showed that the protein expression levels of Wnt1 andβ-catenin in mouse brain tissue were significantly decreased(P<0.01),while the protein expression levels of Cyclin D_(1)and p-GSK-3βwere significantly increased(P<0.01)after LPS injection.Compared with the model group,protein expression levels of Wnt1 andβ-catenin in brain tissue of S.flavescens seed medium and high dose groups were significantly increased(P<0.01),while the protein expression levels of Cyclin D_(1)and p-GSK-3βwere significantly decreased(P<0.01).TUNEL staining results showed that the hippocampal cell apoptosis rate in the model group was significantly increased compared with that of the normal group(P<0.01),while the hippocampal cell apoptosis rate in the S.flavescens seed medium and high dose groups was significantly decreased compared with that of the model group(P<0.01).Conclusion:S.flavescens seed extract can effectively improve the severity of depression in LPS-induced depressed mice,and its molecular mechanism is related to the regulation of neuroinflammation and hippocampal neuronal apoptosis mediated by Wnt/β-catenin signaling pathway.
作者
朱天
王茹
边丽华
李文静
李洁
陈两绵
王智民
高慧敏
郭建友
ZHU Tian;WANG Ru;BIAN Lihua;LI Wenjing;LI Jie;CHEN Liangmian;WANG Zhimin;GAO Huimin;GUO Jianyou(Institute of Psychology,Chinese Academy of Sciences,Beijing 100101,China;University of Chinese Academy of Sciences,Beijing 100049,China;Beijing Health Vocational College,Beijing 101101,China;Institute of Chinese Materia Medica,China Academy of Chinese Medical Sciences,Beijing 100700,China)
出处
《中国实验方剂学杂志》
CAS
CSCD
北大核心
2023年第24期122-129,共8页
Chinese Journal of Experimental Traditional Medical Formulae
基金
国家重点研发计划“中医药现代化研究”专项(2017YFC 1701900)
中国科学院心理研究所自主部署课题(E2CX4115CX)。
