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湖北中医药大学 基础医学院,武汉 430065
黄晓宇,在读硕士,从事中医药防治睡眠疾病研究,E-mail:huangxy1021@stmail.hbtcm.edu.cn
黄攀攀,博士,副研究员,硕士生导师,从事中医药防治睡眠疾病研究,E-mail:panpanhuang@hbtcm.edu.cn
收稿日期:2020-04-08,
网络出版日期:2020-09-27,
纸质出版日期:2020-11-20
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黄晓宇,谢光璟,黄攀攀.天王补心丹加减干预睡眠剥夺大鼠能量代谢机制的生物信息学分析[J].中国实验方剂学杂志,2020,26(22):172-180.
HUANG Xiao-yu,XIE Guang-jing,HUANG Pan-pan.Bioinformatics Analysis on Mechanism of Modified Tianwang Buxindan Intervention on Energy Metabolism in Sleep Deprived Rats[J].Chinese Journal of Experimental Traditional Medical Formulae,2020,26(22):172-180.
黄晓宇,谢光璟,黄攀攀.天王补心丹加减干预睡眠剥夺大鼠能量代谢机制的生物信息学分析[J].中国实验方剂学杂志,2020,26(22):172-180. DOI: 10.13422/j.cnki.syfjx.20202239.
HUANG Xiao-yu,XIE Guang-jing,HUANG Pan-pan.Bioinformatics Analysis on Mechanism of Modified Tianwang Buxindan Intervention on Energy Metabolism in Sleep Deprived Rats[J].Chinese Journal of Experimental Traditional Medical Formulae,2020,26(22):172-180. DOI: 10.13422/j.cnki.syfjx.20202239.
目的
2
使用基因芯片测序技术测定睡眠剥夺大鼠模型被天王补心丹干预前后,能量代谢相关差异基因功能表达谱的变化,为睡眠剥夺的防治提供可能思路及理论依据。
方法
2
采用多平台水环境法对大鼠进行睡眠剥夺造模后随机分为天王补心丹组和模型组,天王补心丹组按照20 g·kg
-1
的剂量灌服天王补心丹水煎剂,模型组给予等体积纯水,连续灌胃14 d。取肝、心、下丘脑为样本,高通量测序获取差异基因,采用基因本体(GO)数据库与京都基因与基因组百科全书数据库(KEGG)通路富集分析,并构建与lncRNA的共表达网络,应用实时荧光定量聚合酶链式反应(Real-time PCR)检测其中3个差异性显著的能量代谢关键基因神经肽Y(NPY),双特异性磷酸酶1/丝裂原活化蛋白激酶磷酸酶1(DUSP1/MKP-1),
α
-
L
-艾杜糖醛酸酶(IDUA)的mRNA表达水平。
结果
2
得到321个差异基因,其中表达上调231个,下调90个,主要促进脂代谢、糖代谢与蛋白代谢进程;参与纤维蛋白原、维生素B
6
与星形胶质细胞源性神经营养因子(MANF)的合成表达;涉及丝裂原活化蛋白激酶(MAPK),p53基因(p53),环磷酸腺苷(cAMP)等信号通路。与模型组比较,天王补心丹组IDUA表达水平明显增加(
P
<
0.05),NPY和DUSP1表达水平显著降低(
P
<
0.01)。
结论
2
天王补心丹从多方面干预睡眠剥夺大鼠的能量代谢机制,通过下调NPY,DUSP1的mRNA表达水平,可能激活参与p38 MAPK信号通路,影响脂质代谢。
Objective
2
To detect the changes of functional expression profile of energy metabolism related differential genes in sleep deprived rats before and after intervention by Tianwang Buxindan by microarray sequencing technology, so as to provide possible ideas and theoretical basis for the prevention and treatment of sleep deprivation.
Method
2
The rats were randomLy divided into two groups: the Tianwang Buxindan group and the model group. The Tianwang Buxindan group was given the decoction of Tianwang Buxindan at the dose of 20 g·kg
-1
, and the model group was given the pure water of equal volume for 14 days. Taking liver, heart and hypothalamus as samples, high-throughput sequencing was used to obtain differential genes. Gene Ontology(Go)classification and kyoto Encyclopedia of Genes and Genomes (KEGG)pathway enrichment analysis were used to construct a co expression network with lncrna. Real-time fluorescence quantitative polymerase chain reaction(Real-time PCR)was used to detect the mRNA expression levels of neuropeptide Y(NPY), bispecific phosphatase 1/mitogen-activated protein kinase phosphatase-1(DUSP1/MKP-1)and alpha-
L
-iduronidase(IDUA), three key genes with significant differences in energy metabolism.
Result
2
The 321 differentially expressed genes were obtained, 231 of which were up-regulated and 90 down regulated, which mainly promoted the process of lipid metabolism, glucose metabolism and protein metabolism, participated in the synthesis and expression of fibrinogen, vitamin B6 and mesencephalic astrocyte-derived neurotrophic factor(MANF), and involved mitogen activated protein kinases(MAPK), p53 gene(p53), cyclic adenosine monophosphate(cAMP) and other signal pathways. Compared with the model group, the expression of IDUA significantly increased in the Tianwang Buxindan group (
P
<
0.05), but decreased significantly in NPY and DUSP1(
P
<
0.01).
Conclusion
2
Tianwang Buxindan can interfere with the energy metabolism mechanism of sleep deprived rats in many ways. By down regulating the mRNA expression level of NPY and DUSP1 genes, it may activate the p38 MAPK signal pathway and affect the lipid metabolism.
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