ABSTRACT
Objective:to explore the mechanism of modified Tianwang Buxindan in improving abnormal glucose and lipid metabolism in mice with chronic sleep deprivation from the signal pathway of orexin A/ orexin receptor 1（OX1R）. Method:The 50 6-week-old male C57BL/6 mice were randomly divided into blank group ， model group ， estazolam group and Tianwang Buxindan low and high dose groups ，for ten mice of each group. Except the blank group， rats were deprived of sleep for 8 weeks by the method of multi-platform water environment. In the last 4 weeks， Tianwang Buxindan （8.5，17 g·kg-1）and estazolam solution（9.1 mg·kg-1）were given to the stomach， and the blank group and the model group were fed with pure water of the same volume. The food intake and body weight of mice were measured twice a week， on the 49th day， blood samples were collected from the tail vein for glucose tolerance test （GTT），on the 52nd day for insulin tolerance test（ITT）， was used to detect the expression of total cholesterol （TCH）， triglyceride（TG）and free fatty acid（FFA）in serum， and enzyme-linked immunosorbent assay（ELISA）was used to detect the expression of orexin A in serum and hypothalamus. Real-time fluorescence quantitative polymerase chain reaction（Real-time PCR）and Western blot were used to detect the mRNA and protein expression of OX1R in hypothalamus. Result:After administration， the food intake of mice in each group was different， compared with the blank group， the body weight of model group was significantly reduced（P<0.05）， the glucose tolerance was significantly abnormal， and the TCH， TG， FFA values were significantly increased（P<0.01）. The expression of orexin A in serum and hypothalamus increased significantly（P<0.01）， and the mRNA and protein expression levels of OX1R in hypothalamus increased significantly（P<0.01）. Compared with the model group， the body weight of each group of Tianwang Buxindan was significantly increased（P<0.05）， with better glucose tolerance and insulin sensitivity， TCH， TG， FFA values were significantly reduced（P<0.05，P<0.01）， accompanied by serum and the expression of orexin A in the hypothalamus was significantly decreased（P<0.05，P<0.01）， the mRNA and protein expression levels of OX1R were significantly decreased（P<0.05，P<0.01）. Conclusion:Tianwang Buxindan can protect mice from abnormal glucose and lipid metabolism induced by chronic sleep deprivation， and its mechanism may be related to the down-regulation of orexin A/OX1R signal expression.
ABSTRACT
Objective: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: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: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: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.
ABSTRACT
Objective:By studying the effects of Tianwang Buxindan on sleep quality， cognitive function， inflammatory factors and immune-related gene expression in sleep deprivation model rats， explore the effect of Tianwang Buxindan on the learning and memory process under sleep deprivation and its anti-inflammatory effects possible mechanism. Method:The 40 male SPF rats were used to simulate the sleep deprivation model by multi-platform water environment method， and were randomly divided into model group， Tianwang Buxindan group （20 g·kg-1） and estazolam group （0.1 mg·kg-1）， set up a normal group， 10 in each group. A total of 4 weeks of sleep deprivation modeling was performed， and drug intervention was performed 2 weeks later. The model group and the blank group were given equal volumes of pure water. Electroencephalogram （EEG） evaluation of modeling and analysis of sleep structure and quality of rats， Morris water maze positioning navigation and space exploration experiment analysis of learning and memory ability of rats， application of enzyme-linked immunosorbent assay （ELISA） was used to detect the serum inflammatory factor interleukin-1β （IL-1β）， tumor necrosis factor-α （TNF-α）， monocyte chemokine-1 （MCP-1） expression， Real-time fluorescence quantitative polymerase chain reaction （Real-time PCR） was used to detect the mRNA levels of EB virus inducible gene 3 （EBI3）， extracellular signal-regulated kinase 5 （ERK5）， and p21 activated protein kinase 4 （PAK4）. Result:The sleep deprivation model was successfully built. Compared with blank group， the total sleep time， total duration of slow wave sleep and the duration of the first and second phases of slow wave sleep in the model group were significantly shortened （P<0.01）. The incubation period on the upper platform， the total swimming distance and the time to reach the original platform for the first time increased significantly， while the number of times to cross the platform and the target quadrant significantly decreased （P<0.05，P<0.01）. The expression levels of IL-1β，TNF-α and MCP-1 increased significantly （P<0.01）， the mRNA expression levels of EBI3， ERK5 and PAK4 in the hypothalamus of the model group decreased significantly （P<0.01）. Compared with the model group， the sleep quality of the rats in the Tianwang Buxindan group was significantly improved. The total sleep time， the total duration of slow wave sleep and the duration of the first phase of slow wave sleep were significantly increased （P<0.01）. The incubation period on the platform， the total swimming distance and the time to reach the original platform for the first time are shortened， the number of times to cross the platform and the target quadrant time are extended （P<0.05，P<0.01）， IL-1β， TNF-α， MCP-1 expression levels were significantly reduced （P<0.05，P<0.01）， mRNA expression levels of EBI3， ERK5 and PAK4 in rat hypothalamus were significantly increased （P<0.05，P<0.01）. Conclusion:Tianwang Buxindan can improve the sleep quality and learning and memory ability of sleep deprivation model rats， which may be related to the increase of the expression level of related inflammatory factors and its anti-inflammatory effect.
ABSTRACT
Objective:To explore the mechanism of modified Tianwang Buxindan on oxidative damage of Trx system in parachlorophenylalanine (PCPA) insomnia model rats. Method:Sixty male SD rats were randomly divided into blank group and model group. Insomnia model was prepared through intraperitoneal injection with PCPA (150 mg·kg-1). The discontinuous injection lasted for 7 d. After successful modeling, the rats were divided into model group (the same volume of normal saline), low, medium, high-dose Tianwang Buxindan groups (8.8, 17.6, 35.2 g·kg-1·d-1) and estazolam group (0.1mg·kg-1·d-1), with 10 in each group. Autonomous activity video was used to detect circadian activity rhythm. Transmission electron microscopy (TEM) was used to observe supra chiasmatic nucleus (SCN) morphology and Organelle integrity. Enzyme-linked immunosorbent assay (ELISA) was used to detect the expressions of superoxide dismutase (SOD), malondialdehyde (MDA) and glutathione peroxidase (GSH-Px) in serum. Expressions of Trx2, TrxR2 in SCN cells were detected by immunofluorescence (IF) and Western blot. Result:Compared with blank group, the activity rhythm of model group was irregular, the activity time increased (PPPPPPPPConclusion:The effect of modified Tianwang Buxindan on insomnia model rats is related to the regulation of Trx2 and TrxR2 protein expressions in Trx system.