摘要
目的:基于高通量测序技术(RNA-Seq)探讨毛蕊花糖苷抗抑郁作用机制,探讨可能作用靶点及信号通路。方法:将40只C57BL/6小鼠随机分为空白组、模型组、氟西汀组及毛蕊花糖苷组,每组10只。除空白组外,其他3组均采用慢性不可预知温和应激(CUMS)结合孤养方式构建抑郁模型,于造模第2周分别给予生理盐水、氟西汀(20 mg·kg^(-1))、毛蕊花糖苷(60 mg·kg^(-1))灌胃,边造模边给药持续3周。通过行为学检测评估小鼠抑郁样状态;酶联免疫吸附测定法(ELISA)检测小鼠血清中主要神经递质及炎性因子含量;小鼠伏隔核及结肠进行mRNA高通量测序,筛选差异表达基因,进行通路富集分析,实时荧光定量聚合酶链式反应(Reat-time PCR)检测伏隔核组织中主要差异基因谷氨酸脱羧酶1(Gad1)、溶质载体家族32成员1(Slc32a1,又名VGAT)、脑源性神经营养因子(BDNF)的mRNA表达。结果:与空白组比较,模型组焦虑、抑郁样行为增加,学习记忆能力显著降低(P<0.01);血清中神经递质含量明显降低(P<0.05);促炎性因子含量明显升高(P<0.05);伏隔核组织中Gad、Slc32a1(VGAT)的mRNA表达量明显下降(P<0.05),BDNF的mRNA表达量明显升高(P<0.05)。与模型组比较,毛蕊花糖苷组小鼠焦虑抑郁样行为显著缓解(P<0.01);神经递质含量明显增加(P<0.05),促炎性因子明显降低(P<0.05);伏隔核组织中Gad、Slc32a1(VGAT)的mRNA表达量明显升高(P<0.05),BDNF的mRNA表达量显著下降(P<0.01)。高通量测序共筛选出伏隔核差异表达基因48个,结肠差异表达基因43个。伏隔核差异基因主要集中在神经活性配体-受体相互作用、γ氨基丁酸(GABA)能突触、突触小泡循环等通路;结肠差异基因主要富集在GABA能突触、突触小泡循环、环磷酸腺苷(cAMP)等信号通路。结论:毛蕊花糖苷具有明显抗抑郁作用。其抗抑郁功效可能与单胺类神经递质增加、促炎因子减少以及通过增加GABA恢复神经递质
Objetive:To investigate the mechanism of antidepressant effect of verbascoside based on high-throughput sequencing technology(RNA-Seq),and to explore the possible targets and signaling pathways.Methods: Forty C57BL/6 mice were randomly divided into control group,model group,fluoxetine group and verbascoside group,10 mice in each group. Except for the control group,all the other three groups were constructed with chronic unpredictable mild stimulation(CUMS)combined with solitary feeding for four weeks.Control group,fluoxetine group and verbascoside group were administered by gavage once daily for three weeks during the second week of modeling. The mice were assessed by sugar-water preference test,forced swimming test,open field test,elevated cross maze test,and water maze test. Enzyme linked immunosorbent assay(ELISA)was performed to detect the levels of major neurotransmitters and inflammatory factors in mice serum,and mRNA high-throughput sequencing was performed in the nucleus accumben and colon to screen differentially expressed genes and perform pathway enrichment analysis. Real-time polymerase chain reaction(Real-time PCR)was used to detect the mRNA expression of Gad,Slc32a1(VGAT)and brain-derived neurotrophic factor(BDNF)in nucleus accumbens. Result: Compared with the control group,the anxiety and depression-like behaviors in the model group increased,while the learning and memory ability decreased significantly. The content of neurotransmitter in serum decreased significantly. The content of pro-inflammatory factors increased significantly. The mRNA expression of Gad and Slc32a1(VGAT)in nucleus accumbens decreased significantly,while that of BDNF increased significantly. Compared with the model group,the anxiety and depression-like behavior of mice in verbascoside group was significantly relieved. The neurotransmitter content increased significantly,and the pro-inflammatory factors decreased significantly. The mRNA expression of Gad and Slc32a1(VGAT)in nucleus accumbens increased significantly,while that o
作者
薛晓燕
潘瑾
施会山
王墉
吴江
高志礼
毛前程
钟桦林
马婷
马柯
XUE Xiaoyan;PAN Jin;SHI Huishan;WANG Yong;WU Jiang;GAO Zhili;MAO Qiancheng;ZHONG Hualin;MA Ting;MA Ke(Shandong University of Traditional Chinese Medicine(TCM),Jinan 250355,China;Shandong University of TCM,Shandong Co-Innovation Center of Classic TCM Formula,Jinan 250355,China)
出处
《中国实验方剂学杂志》
CAS
CSCD
北大核心
2022年第14期148-157,共10页
Chinese Journal of Experimental Traditional Medical Formulae
基金
国家自然科学基金青年基金项目(81903948)。
