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This study aimed to investigate the mechanism of Jiu Wei Bu Xue Oral Liquid on insomnia rats combining the methods of network pharmacology, molecular docking and experimental verification. UPLC-Q-TOF-MS/MS method and TCMIP, TCMSP databases were used to collect the ingredients and targets of Jiu Wei Bu Xue Oral Liquid. Protein-protein interactions and network analysis were performed to screen the key network targets and putative active ingredients of Jiu Wei Bu Xue Oral Liquid in treatment of insomnia, and then following by biological function and KEGG pathway analysis. Then binding ability for key network targets and putative active ingredients were predicted with molecular docking. The prediction targets were validated in para-chlorophenylalanine (PCPA) induced insomnia rats with administration of Jiu Wei Bu Xue Oral Liquid (2, 4, 8 mL·kg-1) for 7 days. Pentobarbital sodium induced sleeping test were performed to evaluate the synergistic sleep-aiding effect of Jiu Wei Bu Xue Oral Liquid. Then glutamic acid (Glu), γ-aminobutyrate (GABA) content and glutamate decarboxylase 1 (GAD67) activity in hypothalamus or hippocampus were evaluated, and the expressions of GAD67, γ-aminobutyric acid receptor subunit α1 (GABRA1) and γ-aminobutyric acid receptor subunit β2 (GABRB2) in hippocampus were detected by qRT-PCR and Western blot methods. Animal experiments were approved by the Institutional Committee on Animal Care of Guangxi Institute of Chinese Medicine & Pharmaceutical Science (the number of permission: 2022060802). Results showed that 16 key network targets and 16 putative active ingredients were obtained by analyzing the herbs-ingredients-targets network of Jiu Wei Bu Xue Oral Liquid in treatment of insomnia. Network pharmacology and molecular docking all indicated these active ingredients, for example atractylenolide Ⅲ, showed better binding ability with GABRA1 and GABRB2. Animal study indicated that, compared to PCPA-induced insomnia model, Jiu Wei Bu Xue Oral Liquid remarkably shortened the sleeping latency and increased the sleeping duration, increased GAD67 activity and the production of GABA in hippocampus of insomnia rats, as well as the expressions of GAD67, GABRA1 and GABRB2, while decreased Glu content in hypothalamus, leading to decreasing of Glu/GABA ratio and recovery of Glu-GABA balance. These results indicated that Jiu Wei Bu Xue Oral Liquid improved insomnia symptoms and helped maintain the Glu-GABA balance within hypothalamus and hippocampus, and reduced the excitatory neurotoxicity within brain. The mechanism may due to the elevation of GAD67 expression and enzyme activity, and the enhancement of type-A GABA receptor (GABAAR)-mediated neurons inhibition.
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ObjectiveTo investigate the effect and mechanism of total flavones of Spatholobi Caulis （TFSC） against depression in rats. MethodThe fifty KM mice were randomly divided into the normal group and high-， medium-， and low-dose （1， 0.5， 0.25 g·kg-1） TFSC groups and gavaged with the corresponding drugs for 12 successive days. One hour after the last administration， the immobility time in forced swimming test and tail suspension test was recorded. The SD rats were randomly divided into the normal group， model group， fluoxetine （5 mg·kg-1） group， and high- and low-dose （1， 0.25 g·kg-1） TFSC groups. Following the exposure of rats to two different kinds of stimuli daily for inducing chronic unpredictable stress， they were administered with the corresponding drugs for 21 d. After the experiment， the levels of serum neurotransmitters and inflammatory factors in rats were detected by enzyme-linked immunosorbent assay （ELISA）. The changes in hippocampal neurons of rats were observed by hematoxylin-eosin （HE） and Nissl staining. The mRNA expression levels of nuclear factor-κB （NF-κB） and tumor necrosis factor-α （TNF-α） in the hippocampus of rats were detected by real-time fluorescence quantitative polymerase chain reaction （Real-time PCR）， and the protein expression levels of cAMP-response element binding protein （CREB）， phosphorylated CREB （p-CREB）， and brain-derived neurotrophic factor （BDNF） in hippocampal tissues by Western blot. ResultCompared with the normal group， TFSC significantly shortened the immobility time of mice in tail suspension and swimming tests （P<0.05）. Compared with the normal group， the model group exhibited reduced sucrose intake and wilderness activity （P<0.01）， decreased 5-HT， DA， NE （P<0.05， P<0.01）， MAO， IL-6， TNF-α （P<0.05， P<0.01）， damaged neurons， increased mRNA levels of TNF-α and NF-κB （P<0.01）， and down-regulated BDNF and CREB protein expression （P<0.05）. Compared with the model group， TFSC significantly enhanced sucrose intake and wilderness activity of rats （P<0.05）， increased the serum 5-HT， DA and NE （P<0.05， P<0.01）， and decreased the serum MAO， IL-6， and TNF-α （P<0.05， P<0.01） as well as NF-κB and TNF-α mRNA expression （P<0.01）， up-regulated the protein expression levels of BDNF and CREB （P<0.01）， and improved the pathological symptoms of hippocampus. ConclusionTFSC improved the hippocampal neurons of rats via CREB/BDNF signaling pathway and reduced depressive pathological damage， thus relieving depression.
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Objective:To investigate the effects and mechanism of Gecko extract for treatment of depression in rats. Method:The depression rats were induced by intraperitoneal injection of reserpine （0.5 mg·kg<sup>-1</sup>）. The successfully modeled rats were randomly divided into model group， fluoxetine group （1.8 mg·kg<sup>-1</sup>）， high dose and low dose groups of Gecko extract （12， 6 g·kg<sup>-1</sup>）. The rats were given corresponding dose of drugs once a day for 10 days. After administration， the levels of neurotransmitters and inflammatory factors in serum and prefrontal cortex of rats were detected by enzyme-linked immunosorbent assay （ELISA）. The cell changes in hippocampal tissues were observed by hematoxylin-eosin （HE） staining. The mRNA levels of interleukin-6 （IL-6）， nuclear factor-<italic>κ</italic>B （NF-<italic>κ</italic>B）， and tumor necrosis factor （TNF-<italic>α</italic>） in the hippocampus of rats were detected by Real-time fluorescence quantitative polymerase chain reaction （Real-time PCR）. The protein levels of Toll-like receptor 4 （TLR4） and NF-<italic>κ</italic>B in hippocampal tissues of rats were detected by Western blot. Result:Compared with the normal group， Gecko extract significantly shortened the immobility time of tail suspension and swimming in mice. Compared with model group， Gecko extract significantly reduced blepharoptosis and retention time in circles for the rats （<italic>P</italic><0.05）， increased the levels of 5-hydroxytryptamine （5-HT） and dopamine （DA） in serum （<italic>P</italic><0.05）， decreased the levels of Monoamine oxidase （MAO）， IL-6， and TNF-<italic>α</italic> in serum （<italic>P</italic><0.05） and prefrontal cortex （<italic>P</italic><0.05）， decreased the mRNA levels of inflammatory cytokines IL-6， NF-<italic>κ</italic>B and TNF-<italic>α</italic> and the protein expressions of TLR4 and NF-<italic>κ</italic>B in the hippocampus of rats （<italic>P</italic><0.05，<italic> P</italic><0.01）， and improved the pathological symptoms of the hippocampus. Conclusion:Gecko extract can significantly alleviate the pathological damage of depression and improve the symptoms of depression， and its mechanism may be due to inhibiting TLR4/NF-<italic>κ</italic>B signaling pathway and reducing the expression of NF-<italic>κ</italic>B， IL-6 and other inflammatory factors in the hippocampus of rats.
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Depression is a kind of mental disease with main symptoms of low mood and lack of pleasure, which seriously endangers human health. An appropriate depressive animal model is of great significance for the study of depression and new antidepressant drugs, while the suitable selection and matching of experimental animals, modeling methods and evaluation indexes are critical to eva-luate the scientificity and effectiveness of the depressive animal model. The study advance of depressive animal models in the aspects of experimental animal selection, modeling principle and method, characteristics, evaluation indexes and their application in traditional Chinese medicine are summarized through the systematic review of relevant literatures in PubMed, CNKI and other databases. The depressive animal modeling methods utilized in recent studies include stress, glucocorticoid induction, reserpine induction, lipopolysaccharide induction, surgical modeling, gene knockout, joint application modeling methods. Stress method is better to simulate the depressive symptoms of clinical patients, whereas there are some deficiencies, such as long modeling time and large cost. The depressive animal models induced by glucocorticoid, reserpine and lipopolysaccharide have the advantages of short modeling time and good controllability, but with a poor reliability. The pathogenesis of surgical modeling is highly matched with that of clinical depressive patients, whereas it has the defect of long postoperative recovery period. Gene knockout models can be used to study the precise role of specific genes in depression. However, its applicability may be restricted in studies on depression. The joint application modeling method can improve its reliability and accuracy, and attracts more and more attention. This paper provides a reference for the selection of animal models in future studies of pathological mechanism of depression, and screening and evaluation of antidepressant drugs.
Subject(s)
Animals , Humans , Antidepressive Agents , Therapeutic Uses , Depression , Disease Models, Animal , Medicine, Chinese Traditional , Mental Disorders , Drug Therapy , Reproducibility of ResultsABSTRACT
Objective: To observe the effect of an active fraction of Polyrhachis vicina (AFPV) on systemic lupus erythematosus (SLE) and its possible mechanism based on animal and cell models. Method: Totally 60 SD rats were randomly divided into normal control group, model group, prednisone acetate group (5 mg·kg-1), and high, medium and low-dose AFPV groups (400, 200, 100 mg·kg-1). SLE model was replicated with bovine serum albumin-Freund's complete (incomplete) adjuvant. Arthus reaction was observed to study the effect of AFPV on the diameter of back skin redness in rats with SLE. The expressions of anti-double-stranded DNA (dsDNA) antibody, complements 3 (C3), complement 4 (C4), immunoglobulin M (IgM), interleukin-1 (IL-1), interleukin-6 (IL-6), interleukin-31 (IL-31) and interleukin-33 (IL-33) in serum were detected by enzyme-linked immunosorbent assay. CD4+T cells were isolated from the spleens of MRL/lpr and C57BL/6J mice at the age of 16 to 18 weeks by immunomagnetic beads method. The expressions of miR-200a and miR-155 and the levels of zinc-finger-enhancer binding protein 1(ZEB1) and suppressor of cytokine signaling1(SOCS1) in CD4+T cells were observed to explore the effect of AFPV on SLE and its possible mechanism. Result: Compared with the normal group, the diameter of back skin swelling in the model group was significantly increased (PPPPPPP+T cells of MRL/lpr lupus mice. Compared with the model group, the expression of microRNA-200a increased significantly, the expression of microRNA-155 decreased significantly (PPConclusion: AFPV has therapeutic effect on rats with SLE, its mechanism may be related to the regulation of miR-200a/ZEB1 and miR-155/SOCS1.
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The main ingredient of extractable petroleum ether of Polyrhachis vicina Roger (EPPR) is octadecene unsaturated fatty acids. Mounting evidence supports that N-3 polyunsaturated fatty acids can attenuate neuroinflammation, reduce oxidative stress, then protect neurons. In order to explore the effect of EPPR on the inflammatory response of depressed rats, the model of depression was established by chronic unpredictable mild stress (CUMS). Sucrose preference test, forced swimming test were employed to investigate the anti-depressive effect of EPPR in rat. The activation of glial cells and astrocytes in the prefrontal cortex of depressed rats was observed by immunofluorescence. The levels of inflammatory factors were measured by Quantitative Real-time PCR. NF-κB was detected by immunoblotting. EPPR could significantly improve the depressive behavior of rats, decrease NF-κB translocation to the compartment of nucleus, down-regulate the pro-inflammatory cytokines IL-1β, TNF-α and indoleamine 2,3-dioxygenase (IDO) gene expression levels, inhibit the activation of microglia and astrocytes in depressed rats. These results suggest that EPPR could notably ameliorate inflammation induced by chronic stress, and the protective effect might be linked to the regulation of NF-κB p65.