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ObjectiveTo investigate the potential mechanism of Xiao Chaihutang （XCHT） in the treatment of Alzheimer's disease （AD） based on network pharmacology and bioinformatics. MethodThe active components of XCHT and corresponding targets were retrieved from Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform （TCMSP）， and the differentially expressed genes related to AD were searched from Gene Expression Omnibus （GEO）. Thereby， the common targets of XCHT and AD were yielded， followed by Gene Ontology （GO） term enrichment and Kyoto Encyclopedia of Genes and Genomes （KEGG） pathway enrichment analysis of the common targets. The component-target network and protein–protein interaction （PPI） network were constructed. Furthermore， amyloid β-protein （Aβ）1-40 was used to induce AD in PC12 cells and then the AD cells were intervened with XCHT. Afterward， cell viability was detected by Cell Counting Kit-8 （CCK-8） assay and cell morphology was observed based on 4'，6-diamidino-2-phenylindole （DAPI） staining. Cell membrane potential was determined and apoptosis was detected by flow cytometry， and cellular immunofluorescence detects the expression of B-cell lymphoma-2 （Bcl-2）/Bcl-2-related X protein （Bax）. Moreover， immunofluorescence assay was performed. ResultA total of 190 active components and 41 anti-AD targets of XCHT were screened out. The key components included mairin， quercetin， berberine， protoporphyrin， 24-ethylcholest-4-en-3-one， and β-D-ribofuranoside， and the core targets were sigma non-opioid intracellular receptor 1 （SIGMAR1）， checkpoint kinase 1 （CHEK1）， protein tyrosine phosphatase non-receptor type 6 （PTPN6）， protein kinase C（PRKCH）， inhibitor of nuclear factor kappa B kinase subunit beta （IKBKB）， cathepsin D （CTSD）， cysteine aspartate protease-3 （Caspase-3）， Bax， and Bcl-2-like protein 1 （Bcl-2L1）. The anti-AD targets of XCHT were involved in 302 GO terms （P < 0.05）， particularly the regulation of neuronal cell apoptosis， and 73 KEGG pathways （P<0.05）. The major pathways and biological processes included the apoptosis pathway， virus infection pathway， lipid and atherosclerosis pathway， and cancer-related pathways. In the in vitro experiment， the model group demonstrated the decrease in cell survival rate （P<0.05）， increase in apoptosis rate （P<0.05）， and down-regulation of mitochondrial membrane potential and Bcl-2/Bax ratio compared with the blank control. Compared with the model group， XCFT group showed the increase in cell survival rate （P<0.05）， decrease in apoptosis rate （P<0.05）， and up-regulation of mitochondrial membrane potential and Bcl-2/Bax ratio. ConclusionBased on network pharmacology， this study reveals the multi-component， multi-target， and multi-pathway characteristics of XCHT in the treatment of AD， laying a foundation for further research on the material basis and mechanism of this prescription.
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Objective::To predict the action targets of anti-lung cancer active ingredients of Xiao Chaihutang, in order to explore the " multi-components, multi-targets and multi-pathways" mechanism using network pharmacology. Method::The active ingredients of Xiao Chaihutang that obtained through Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform(TCMSP), traditional Chinese medicine integrative database for herb molecular mechanism analysis(TCMID) and literature were used to predict the targets by the reversed pharmacophore matching method.To screen out optimization targets, we chose elbow point analysis by using self-developed software TCMKD1.0, and screened out lung cancer-related targets by searching databases, such as Therapeutic Target Database (TTD), Online Mendelian Inheritance in Man(OMIM) and GeneCards, and reviewing literatures.Then components-target network, protein-protein interaction network and targets-pathways network were constructed.The pathway information was acquired with STRING.The Cytoscape 3.6 software was used to construct the ingredients-targets-pathways network of Xiao Chaihutang. Result::The 162 active components in Xiao Chaihutang were obtained, involving 71 anti-lung cancer targets and 11 related pathways.Through topological network analysis, 96 important components, such as quercetin, ginsenosideRh2, formononetin and β-sitosterol were obtained, 28 key targets, such as epidermal growth factor receptor (EGFR), vascular endothelial growth factor (KDR), cysteine protease-3 (Caspase-3), mitogen-activated protein kinase1(MAPK1), hepatocyte growth factor (MET) were received, and 61 core pathways, such as non-small cell lung cancer, small cell lung cancer, ErbB signaling pathway, VEGF signaling pathway were acquired. Conclusion::The result suggests that the active components of Xiao Chaihutang against lung cancer may include quercetin, ginsenoside Rh2, 6-shogaol, formononetin, β-sitosterol.And the mechanism may be related to ErbB signaling pathway, MAPK signaling pathway and VEGF signaling pathway.This research provides a scientific basis for further elucidation of the anti-lung cancer pharmacological mechanism of Xiao Chaihutang.
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Objective:To study the effect of modified Xiao Chaihutang on the expressions of excitatory amino acid transporters（EAATs） and vesicle glutamate transporters（VGLUTs）in hippocampus of rats with chronic depression， in order to explore the anti-depressant mechanism of modified Xiao Chaihutang based on glutamate transport. Method:A total of 120 SD rats were randomly divided into normal group， model group， and low， middle and high-dose modified Xiaochaihutang groups （6.5， 13， 26 g·kg-1） and riluzole group， with 20 rats in each group．Except normal group， the depression model of rats was prepared through Chronic restraint stress（CRS）. The normal group and the model group were intragastrically （ig） given normal saline. The modified Xiao Chaihutang groups were intragastrically given corresponding herbal drugs （6.5， 13， 26 g·kg-1）， and the Riluzole group was given Riluzole 20 mg·kg-1 through intraoeritoneal injection for 21 days， once a day. Then the depressive behaviors of rats were observed by forced swimming test （FST） and tail suspension test （TST）. The level of glutamic acid （Glu） in rats hippocampus was determined by high performance liquid chromatography （HPLC）. The mRNA expressions of EAAT1， EAAT2 and EAAT3 in hippocampus were detected by Real-time fluorescence quantitative polymerase chain reaction （Real-time PCR）method. Western blot was used to detect the protein expressions of EAAT1， EAAT2， EAAT3， VGLUT1 and VGLUT2 in rat hippocampus tissue. Nissl staining was used to observe the morphology of hippocampal neurons in rats. Immunohistochemical（IHC）S-P method were used to detect the location expressions of EAAT1， EAAT2 and NeuN proteins in rat hippocampal CA1 region tissue. Result:The immobility times in FST and TST were increased significantly（P<0.01）， the mRNA and protein expressions of EAAT1，EAAT2，EAAT3 were decreased significantly （P<0.01）， and as well as the expressions of VGLUT1 and NeuN were decreased significantly（P<0.01）， while the level of Glutamate and the expression of VGLUT2 were increased significantly（P<0.01） in model group， compared with normal group. Compared with model group，the immobility times in FST and TST were decreased significantly（P<0.05， P<0.01）， mRNA and protein expressions of EAAT1，EAAT2，EAAT3 were increased significantly（P<0.01）， and expressions of VGLUT1 and NeuN were increased significantly（P<0.01）. However， the level of Glutamate and the expression of VGLUT2 were decreased significantly（P<0.01）， and the damage of hippocampal neurons in rats was mild in middle and high-dose modified Xiao Chaihutang groups. Conclusion:Modified Xiao Chaihutang has an anti-depressive effect. Its mechanism may be related to its up-regulation of expressions of EAAT1， EAAT2， EAAT3 genes and VGLUT1 protein in the hippocampus of depression model rats.
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Objective:To observe the expression of tumor necrosis factor receptor-associated death domain (TARDD), nuclear transcription factor-κB inhibiting protein α(IκBα)IκB kinase-α (IKKα) and nuclear transcription factor (NF)-κB p65 protein in the NF-κB signaling pathway of synovial tissues of complete Freund's adjuvant (CFA) rats after treatment with Xiao Chaihutang (XCHT). Method:In animal experiments, SPF health adult female Wistar rats were used to prepare the CFA animal model of rats with rheumatoid arthritis with Freund's complete adjuvant and cattle Ⅱ collagen type. According to the random number table, the rats were randomly divided into the normal group, the model group, the low-dose XCHT group, the medium-dose XCHT group, the high-dose XCHT group, and the Tripterygium glucosides group. The drugs were given at 7 d after the model was built. Both normal group and model group were given water for injection,and low-dose XCHT group(5.94 g·kg-1),medium-dose XCHT group(11.88 g·kg-1),high-dose XCHT group(23.76 g·kg-1),Tripterygium glucosides group(0.006 3 g·kg-1) were given corresponding drugs by gavage for three times a day, 2 mL/time. The histopathology of rat ankle joint was observed, and the protein expressions of TARDD,IKKα,IκBα,NF-κB p65 in the NF-κB signaling pathway in synovial tissue of CFA rats were detected by Western blot. Result:With the increase of the dosage of XCHT, the histopathological score of the right posterior ankle joint of the experimental rats was increased. And in the protein expressions of TARDD,IKKα,IκBα,NF-κB p65 in NF-κB signaling pathway in Synovial Tissue of CFA rats, compared with the model group, the statistical results of the low-dose XCHT group showed decreased protein expressions (PPPα, IκB α, NF-κB p65 in the NF-κB signaling pathway were significantly increased (PPα, IκBα, NF-κB p65 key protein expressions in the NF-κB signaling pathway and protein expressions in low-dose XCHT group were obviously lower (PPConclusion:This study shows that as the dose of Xiao Chaihutang increases, it could effectively improve synovitis, and suppress the expressions of key proteins in the inflammatory signaling pathway of NF-κB, thereby preventing inflammation and suppressing bone erosion.
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Objective: To observe the clinical efficacy of Hegan granule in the treatment of non-erosive reflux disease with liver stagnation and spleen deficiency syndrome and its effect on quality of life, 5-hydroxytryptamine (5-HT) and vasoactive intestinal peptide (VIP) level. Method: Patients with non-erosive reflux disease of liver stagnation and spleen deficiency were randomly divided into treatment group (32 cases) and control group (33 cases). The treatment group was orally given Hegan granules after meal, 10 g/bag, 1 bag/time, 2 times/day. The control group was orally given with omeprazole enteric-coated tablets twice daily, 20 mg/grain. Both groups were treated for 8 weeks. The total scores of traditional Chinese medicine (TCM) symptoms, SF-36 health scale scores, 5-HT and VIP levels were observed before and after treatment. Result: The medical efficacy of the two groups was 87.5%and 81.8%in the treatment group and the control group, respectively. The treatment group was superior to control group (PPPPConclusion: Hegan granule has a good clinical efficacy in treating patients with non-erosive reflux disease and liver stagnation and spleen deficiency. It can significantly alleviate the symptoms of TCM, improve the quality of life of patients, and reduce the sensitivity of esophageal viscera.
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Objective: To observe the therapeutic effect of Xiaochaihu granule on acute liver injury (ALI) induced by thioacetamide (TAA) in rats, and to explore the role of transcription factor (NF) -E2 related factor 2 (Nrf2) in the pathway of oxidative damage. Method: SD rats were randomly divided into control group, model group, Xiaochaihu granule low, middle and high dose groups (1, 2, 4 g·kg-1). 250 mg·kg-1 TAA was given to the rats by ip. administration for 2 days to prepare the liver injury model, and from the 3rd day, same amount of double distilled water or different doses of Xiaochaihu granule was given to corresponding groups by ig. administration for 2 weeks. 24 hours after the last administration, liver tissues were taken and stained with hematoxylin-eosin(HE) staining. The activities of alanine aminotransferase (ALT) and aspartate aminotransferase (AST) in serum were measured by colorimetry. The activities of total superoxide dismutase (T-SOD) and malondialdehyde (MDA) in liver tissues were measured by colorimetry. Real-time PCR was used to detect Nrf2 pathway related mRNA expression. Result:As compared with control group, ALT, AST and MDA in model group were significantly increased,while T-SOD was significantly decreased (PPPPPPPConclusion:Xiaochaihu granule may play a therapeutic role in TAA induced ALI in rats by up-regulating the expression of downstream molecules in Nrf2 signaling pathway.
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Objective: To observe the protective effect of Xiao Chaihutang and Wulingsan on residual renal function in patients with maintenance peritoneal dialysis (PD) and investigate its effect on peritoneal fibrosis and microinflammation.Method: The 65 patients with PD who were admitted to our hospital from June 2016 to June 2017 were enrolled in the study, and divided into control group (32 cases) and study group (33 cases) according to the random number table. The control group received routine treatment. The study group received routine treatment+Xiao Chaihutang and Wulingsan. The fasting venous blood was taken before treatment and 3 months after treatment to measure serum creatinine (SCr) and urea nitrogen (BUN). Urine was collected; 24 hour urine volume was recorded; 24 h urine protein (24 h UP) was measured by colorimetry; glomerular filtration rate (eGFR) was calculated; residual renal function (RRF) was expressed with residual renal creatinine clearance. Inflammatory factors were detected by using chemiluminescence, including interleukin-6 (IL-6), interleukin-8 (IL-8), and tumor necrosis factor-α (TNF-α) levels. Serum Janus Kinase (JAK) 2 as well as signal transducer and activator of transcription (STAT) 3 levels were determined by double antibody sandwich enzyme-linked immunosorbent assay (ELISA). E-cadherin and α-smooth muscle actin (α-SMA) levels were determined by Western blot.Result: Before treatment, there was no significant difference in the residual renal function between two groups. After treatment, the residual renal function of the study group was significantly better than that of control group (PPα in study group were lower than those in control group (PPPα-SMA protein was significantly increased after treatment (Pα-SMA protein after treatment.Conclusion: Xiao Chaihutang and Wulingsan can protect the residual renal function of PD patients, and the mechanism may be related to the improvement of peritoneal fibrosis and the reduction of micro-inflammation of the body, showing a high application value.