基于lncRNA/mRNA表达谱探讨百事乐胶囊调控慢性应激抑郁症模型大鼠海马损伤的机制  

Exploration of the Mechanism of Baishile Capsules Regulating Hippocampal Injury in Chronic Stress Depression Model Rats Based on lncRNA/mRNA Expression Profile

作　　者：刘彤彤 侯光菡 蔺晓源[3] 杨蕙[3] 刘检[3] 张熙 王枭冶 王宇红[1] 葛金文[1,5] 孟盼 LIU Tongtong;HOU Guanghan;LIN Xiaoyuan;YANG Hui;LIU Jian;ZHANG Xi;WANG Xiaoye;WANG Yuhong;CE Jinwen;MENG Pan(Science and Technology Innovation Center,Hunan University of Chinese Medicine,Changsha 410208 Hunan,China;The Fourth Hospital of Changsha,Changsha 410006 Hunan,China;Medical Experiment Center,The First Affiliated Hospital of Hunan University of Chinese Medicine,Changsha 410007 Hunan,China;Brain Hospital of Hunan Province,Changsha 410007 Hunan,China;Hunan Academy of Chinese Medicine,Changsha 410006 Hunan,China)

机构地区：[1]湖南中医药大学科技创新中心,湖南长沙410208 [2]长沙市第四医院,湖南长沙410006 [3]湖南中医药大学第一附属医院医学实验中心,湖南长沙410007 [4]湖南省脑科医院,湖南长沙410007 [5]湖南省中医药研究院,湖南长沙410006

出　　处：《中药新药与临床药理》2023年第3期312-321,共10页Traditional Chinese Drug Research and Clinical Pharmacology

基　　金：湖南省自然科学基金优秀青年项目(2020JJ3027);湖南省科技创新人才计划优秀博士后创新人才项目(2020RC2060);长沙市杰出创新青年培养计划项目(kq2009018);长沙市自然科学基金项目(kq2208471)。

摘　　要：目的基于长链非编码RNA(lncRNA)/信使RNA(mRNA)表达谱探讨百事乐胶囊(贯叶金丝桃、人参、姜黄)调控慢性应激抑郁症模型大鼠海马损伤的可能机制。方法将SD大鼠随机分为空白组、模型组及百事乐胶囊高、低剂量组(2.88、0.72 g·kg^(-1)),每组8只;采用28 d慢性温和不可预测性应激联合孤养的方式构建抑郁症大鼠模型,造模同时灌胃给药,每日1次。采用开野试验和新奇摄食试验评价大鼠模型的抑郁样行为;采用ELISA法检测血清中肿瘤坏死因子α(TNF-α)、白细胞介素6(IL-6)的水平;采用高通量芯片测序筛选出组间差异的海马组织lncRNA/mRNA表达谱,预测差异表达的lncRNA靶基因,并取lncRNA、mRNA的差异共集基因;对差异共集基因进行GO功能富集分析和KEGG通路富集分析;采用Western Blot法验证差异性蛋白胰岛素生长因子2(Igf2)、核糖体蛋白L36(Rpl36)的表达情况。结果(1)与空白组比较,模型组大鼠4 min内的水平、垂直活动量均显著降低(P<0.01),摄食潜伏期显著延长(P<0.01),血清TNF-α、IL-6水平显著升高(P<0.01)。与模型组比较,百事乐胶囊高剂量组大鼠的水平、垂直活动量明显增加(P<0.05,P<0.01),摄食潜伏期显著缩短(P<0.01),血清TNF-α、IL-6水平明显降低(P<0.05,P<0.01)。(2)模型组与空白组比较,差异共集基因有152个,其中上调100个,下调52个;百事乐胶囊高剂量组与模型组比较,差异共集基因有150个,其中上调88个,下调62个;百事乐胶囊低剂量组与模型组比较,差异共集基因有119个,其中上调58个,下调61个。Igf2、Rpl36为百事乐胶囊高剂量组的主要逆向调节基因。百事乐胶囊主要调控与生物学过程(突触化学传递)和细胞组分(细胞连接)密切相关的差异共集基因的表达,可能通过影响MAPK信号转导通路、胰岛素信号改善抑郁症模型大鼠的海马损伤。(3)与空白组比较,模型组大鼠海马组织Igf2、Rpl36蛋白表达显著下调(P<0.01)Objective To explore the possible mechanisms of Baishile Capsules (consists of Hyperici Perforati Herba,Ginseng Radix et Rhizoma,Curcumae Longae Rhizoma) regulating hippocampal injury in chronic stress depression model rats based on long non-coding RNA (lncRNA)/messenger RNA (mRNA) expression profiles.Methods The SD rats were randomly divided into blank group,model group and Baishile Capsules high-and lowdose groups (2.88 and 0.72 g·kg^(-1)),with 8 rats in each group.A 28-day chronic mild unpredictable stress combined with solitary rearing was used to construct a rat model of depression,and the model was constructed and administered by gavage once daily.The depression-like behavior of the rat model was evaluated using the open field test and the novelty-suppressed food test;the levels of tumor necrosis factor α (TNF-α)and interleukin 6 (IL-6)in serum were measured by ELISA;the lncRNA/mRNA expression profiles of hippocampal tissue were screened for differences between groups using high-throughput microarray sequencing to predict differentially expressed lncRNA target genes,and the lncRNA,mRNA co-collected genes;GO functional enrichment analysis and KEGG pathway enrichment analysis were performed on the differentially co-collected genes;Western Blot was used to verify the expressions of differentially expressed proteins insulin growth factor 2 (Igf2)and ribosomal protein L36 (Rpl36).Results (1)Compared with the blank group,the horizontal and vertical activity levels within 4 minutes in model group were significantly decreased (P<0.01),the feeding latency was significantly prolonged (P<0.01),and the serum levels of TNF-α and IL-6 were significantly increased (P<0.01).Compared with the model group,the levels of horizontal and vertical activity of rats in high-dose group were significantly increased (P<0.05,P<0.01),the feeding latency was significantly shortened (P<0.01),and the levels of serum TNF-α and IL-6 were significantly decreased (P<0.05,P<0.01).(2) There were 152 genes were differentially co-collected in

关 键 词：百事乐胶囊 抑郁症 长链非编码RNA/信使RNA表达谱 海马损伤 胰岛素生长因子2 核糖体蛋白L36 大鼠 

分 类 号：R285.5[医药卫生—中药学]