摘要
该研究观察抗抑郁中药古方定志小丸（DZ）在体内外对miR-16表达水平的影响,从转录后调控的角度探索定志小丸升高5-羟色胺（5-HT）水平的机制。首先建立慢性不可预知中等强度温和刺激（CUMS）结合孤养抑郁大鼠模型,观察不同剂量定志小丸（600,300,150 mg·kg-1）给药3周后对抑郁大鼠行为学指标的影响,高效液相色谱法（HPLC）观察对海马5-羟色胺（5-HT）含量的影响,PCR和Western blot法观察定志小丸对CUMS大鼠海马miR-16及SERT表达水平的影响。并在体外培养的大鼠原代海马神经元细胞中观察定志小丸（10,100,200,500 mg·L-1）作用48 h后对miR-16沉默和皮质酮/谷氨酸损伤导致的细胞模型中miR-16及SERT mRNA表达水平的影响。结果显示,300,600 mg·kg-1定志小丸可显著改善CUMS大鼠行为学评分,升高大鼠海马内5-HT的水平,同时能够升高大鼠海马区miR-16的表达,降低SERT的表达。同时,在大鼠原代海马神经元细胞中,100,200,500 mg·L-1定志小丸可显著升高miR-16沉默和皮质酮/谷氨酸损伤模型细胞中miR-16的表达水平,并显著降低上述造模引起的细胞内SERT表达水平的升高。综上,笔者认为定志小丸可能通过上调miR-16表达水平,释放其对靶基因SERT的抑制作用,抑制5-HT的重摄取,进而升高脑内5-HT的水平,发挥抗抑郁作用。
This study is to investigate the effect of antidepressant medicine prescription Dingzhi Xiaowan（ DZ） on miR16 expression levels in vitro and in vivo,and to explore the mechanism of DZ elevated levels of 5-HT from the perspective of post transcriptional regulation. Firstly,a chronic unpredictable mild moderate stimulation（ CUMS） combined with solitary rising depression rat model was established,the behavior changes were detected after different doses of DZ（ 600,300,150 mg·kg（-1）） given for 3 weeks,high performance liquid chromatography（ HPLC） was used to detect 5-HT level in hippocampal,PCR method was used to observe the effect of DZ on the expression of SERT mRNA and miR16 in hippocampus of CUMS rat. The effects of DZ（ 10,100,200,500 mg·L（-1）） on the expression of miR16 and SERT mRNA in the cell model induced by miR16 silencing and corticosterone or glutamate injury were observed in primary cultured hippocampal neurons of rats in vitro. It was found that 300 mg·kg（-1） and 600 mg·kg（-1） DZ could significantly improve the behavioral score of CUMS rats,increase the level of 5-HT in hippocampus,and increase the expression of miR16 and decrease the expression of SERT in the hippocampus of rats. At the same time,in primary cultured hippocampal neurons,100,200,500 mg·L（-1） of DZ could significantly increase the expression level of miR16 in miR16 silencing and corticosterone or glutamate injury cell model,and decrease the expression level of SERT significantly. So DZ could inhibit the reuptake of 5-HT by inhibiting the expression of SERT by up regulating the expression level of miR16,and finally increase the level of 5-HT in the brain and exert antidepressant effect.
作者
朱维煜
冯霞
王瑾
路玉盼
董宪喆
刘屏
ZHU Wei-yu;FENG Xia;WANG Jin;LU Yu-pan;DONG Xian-zhe;LIU Ping(Department of Clinical Pharmacology,General Hospital of PLA,Beijing 100853,China;Outpatient Department of the 61st Research Institute of the PLA General Staff Headquarters,Beijing 100041;Department of Graduate,Shanxi University of Traditional Chinese Medicine,Jinzhong 030600,China)
出处
《中国中药杂志》
CAS
CSCD
北大核心
2018年第17期3513-3518,共6页
China Journal of Chinese Materia Medica
基金
国家自然科学基金项目(81302909,81673877)
