In this paper,metabolomics and network pharmacology were used to investigate the bioactive components of Harrisonia perforata and their possible mechanisms of action. Metabolites in the flowers,fruits,branches,leaves and stalks of H. perforata were analyzed by ultra-high performance liquid chromatography-quadrupole-time-of-flight mass spectrometry. Meanwhile,multiple statistical analysis methods including principal component analysis( PCA) and orthogonal partial least squares discriminant analysis( OPLS-DA)were applied to screen and identify differential compounds. With metabolomics method,9 differential compounds were preliminarily identified from leaves and other non-traditional medicinal parts. Subsequently,these compounds were explored by using network pharmacology. With gastrointestinal absorption and drug-likeness as limiting conditions,they were imported into the Swiss ADME,from which 7 compounds with potential medicinal activity were obtained. Then,their targets were predicted by PharmMapper,with Human Protein Targets Only and Normalized Fit Score>0. 9 set as limiting conditions,and 60 standardized potential targets were identified with Uniprot. KEGG( Kyoto encyclopedia of genes and genomes) pathway data was obtained using metascape and the " potential active ingredients-target-pathway" network was constructed with Cytoscape 3. 7. 2. The enrichment analysis of KEGG demonstrated that the 60 targets were enriched in 78 signaling pathways( min overlap: 3,P value cutoff: 0. 01,min enrichment: 1. 5),many of which are related to anti-bacteria,anti-inflammation and anti-virus,such as IL-17 signaling pathway,RIG-I-like receptor signaling pathway and NOD-like receptor signaling pathway. Finally,depending on the clinical activity of H. perforata,the relevant signaling pathways were analyzed through experimental data and literature. Dehydroconiferyl alcohol was reported to have the anti-inflammatory effect and perforamone D to possess the antimycobacterial activity. The KEGG pathway enrichment analysis showed that dehydroconiferyl alcohol could act on the Alzheimer's disease( AD) signaling pathway by targeting CDK5 R1 and BACE1. ACh E inhibitor is the most promising drug to treat AD,while dehydroconiferyl alcohol has been proved to inhibit ACh E according to literature. The experimental results revealed that the extract of leaves of H. perforata can effectively inhibit the growth of Staphylococcus aureus. These are consistent with the enrichment analysis results of KEGG. This study explored the bioactive components and pharmacodynamics of the leaves of the H. perforata,laying a theoretical foundation for its in-depth development and rational application.
Amyloid Precursor Protein Secretases ; Aspartic Acid Endopeptidases ; Drugs, Chinese Herbal/pharmacology* ; Humans ; Metabolomics ; Simaroubaceae

OBJECTIVETo study the changes of excitatory and inhibitory amino acid content in hippocampus of epileptic rats treated with volatile oil of A. tatarinowii, and explore the possible antiepiletic mechanism.

METHODThe volatile oil was extracted through Supercritical-CO2 Fluid Extraction (SFE-CO2), and epileptic models were built up by kainic acid (KA) lateral ventricle injection. The content of amino acid in hippocampus of epileptic rats treated with volatile oil was calculated.

RESULTThe content of GABA increased and Glu decreased prominently (P < 0.05) after volatile oil 35 mg x kg(-1) intraperitoneal injection.

CONCLUSIONThe volatile oil of A. tatarinowii can modulate the balance of excitatory and inhibitory amino acid in epileptic rats, thereby exerting its antiepileptic effect.

Acorus ; chemistry ; Animals ; Anticonvulsants ; pharmacology ; Aspartic Acid ; metabolism ; Epilepsy ; chemically induced ; metabolism ; Glutamic Acid ; metabolism ; Hippocampus ; metabolism ; Kainic Acid ; Male ; Oils, Volatile ; isolation & purification ; pharmacology ; Plants, Medicinal ; chemistry ; Rats ; Rats, Sprague-Dawley ; gamma-Aminobutyric Acid ; metabolism

The purpose of this study was to determine whether esterification of dehydroepiandrosterone with aspartate (DHEA-aspartate) could reduce peroxisomal proliferation induced by DHEA itself, without loss of antiosteoporotic activity. Female Sprague-Dawley rats were ovariectomized, then DHEA or DHEA-aspartate was administered intraperitoneally at 0.34 mmol/kg BW 3 times a week for 8 weeks. DHEA-aspartate treatment in ovariectomized rats significantly increased trabeculae area in tibia as much as DHEA treatment. Urinary Ca excretion was not significantly increased by DHEA or DHEA-aspartate treatment in ovariectomized rats, while it was significantly increased by ovariectomy. Osteocalcin concentration and alkaline phosphatase activity in serum and cross linked N-telopeptide type I collagen level in urine were not significantly different between DHEA-aspartate and DHEA treated groups. DHEA-aspartate treatment significantly reduced liver weight and hepatic palmitoyl-coA oxidase activity compared to DHEA treatment. DHEA-aspartate treatment maintained a nearly normal morphology of peroxisomes, while DHEA treatment increased the number and size of peroxisomes in the liver. According to these results, it is concluded that DHEA-aspartate ester has an inhibitory effect on bone loss in ovariectomized rats with a marked reduction of hepatomegaly and peroxisomal proliferation compared to DHEA.
Adjuvants, Immunologic/pharmacology* ; Adjuvants, Immunologic/metabolism ; Adjuvants, Immunologic/chemistry ; Animal ; Aspartic Acid/pharmacology* ; Aspartic Acid/metabolism ; Aspartic Acid/chemistry ; Biological Markers ; Calcium/urine ; Calcium/blood ; Disease Models, Animal ; Esterification ; Fatty Acid Desaturases/metabolism ; Female ; Injections, Intraperitoneal ; Lipoproteins, HDL Cholesterol/blood ; Lipoproteins, LDL Cholesterol/blood ; Liver/enzymology ; Liver/drug effects ; Organ Weight ; Osteoporosis/pathology ; Osteoporosis/metabolism* ; Osteoporosis/drug therapy* ; Ovariectomy* ; Peroxisomes/metabolism* ; Prasterone/pharmacology* ; Prasterone/metabolism ; Prasterone/chemistry ; Rats ; Rats, Sprague-Dawley ; Tibia/pathology ; Tibia/metabolism ; Triglycerides/blood

OBJECTIVETo research the content changes of excitatory neurotransmitter and inhibitory neurotransmitter in corpus striatum of rats after single-used borneol and combining it with diazepam in hope of comprehending the activity of borneol on central nervous system and to observe whether borneol could increase the penetration of other drugs into the brain.

METHODThe content of four amino acids neurotransmitters in corpus striatum of rats were sampled by brain microdialysis technology at different time after administration and were determined by RP-HPLC which involved pre-column derivation with orthophthaladehyde (OPA), using phosphate gradient elution and fluorescence detection to detect the content of excitatory neurotransmitter aspartate (Asp), glutamate (Glu) and inhibitory neurotransmitter glycine (Gly), gamma-aminobutyric acid (GABA) in standards and samples and carry on statistical analysis.

RESULTThe content of both Gly and GABA in corpus striatum of rats with borneol increased significantly, compared with diazepam group (P < 0.05), while Asp and Glu showed no significant difference.

CONCLUSIONBorneol can improve permeability of diazepam through BBB.

Animals ; Aspartic Acid ; analysis ; Blood-Brain Barrier ; Bornanes ; administration & dosage ; pharmacology ; Corpus Striatum ; chemistry ; drug effects ; Diazepam ; administration & dosage ; pharmacology ; Glutamic Acid ; analysis ; Glycine ; analysis ; Male ; Neurotransmitter Agents ; analysis ; Rats ; Rats, Sprague-Dawley ; gamma-Aminobutyric Acid ; analysis

BACKGOUND: This study was purposed to evaluate the effects of Mg2+ pretreatment on cerebral ischemic injury in cats. METHODS: Global cerebral ischemia was induced by ligation of both innominate arteries following ligation of inferior vena cava under lowered mean blood pressure for 20 minutes followed by 3 hrs of reperfusion. Ten cats were divided into 2 groups: Group 1 (n=5) is the control group, for group 2 (n=5) (Mg2+ group), the animals were pretreated with 90 mg/kg of Mg2+ intravenously before subjected to ischemia. RESULTS: Phosphocreatine/inorganic phosphate (Pcr/Pi) and pH decreased after ischemia and did not recovered during reperfusion. And there were no significant differences between the two groups. The ratios of lactate/N-acetyl aspartate (Lac/NAA) and lactate/creatine (Lac/Cr) increased after ischemia and not recovered during reperfusion. But the ratios were higher for the group 2 than the group 1 during reperfusion (p<0.05). For the Mg2+ group, blood pressure during reperfusion was lower than the control group. CONCLUSIONS: Mg2+ intravenous pretreatment had no protective effect on this global cerebral ischemia animal model. Even it deteriorated brain energy metabolism by lowering blood pressure.
Animals ; Aspartic Acid ; Blood Pressure ; Brachiocephalic Trunk ; Brain Ischemia ; Brain* ; Cats* ; Energy Metabolism* ; Hydrogen-Ion Concentration ; Ischemia ; Ligation ; Metabolism ; Models, Animal ; Pharmacology ; Reperfusion ; Vena Cava, Inferior

OBJECTIVETo explore the effect of Panax notoginseng saponin (PNS) on the activity and content of beta-secretase in the brain of senescence accelerated mouse-prone 8 (SAMP8) mice with Alzheimer's disease.

METHODSTotally 32 SAMP8 mice were randomly divided into the normal control group, the high dose PNS group (200 mg/kg), the low dose group (100 mg/kg), and the huperzine A group (0.3 mg/kg), 8 in each group. Equal volume of double distilled water was given to those in the normal control group. All medication was given by gastrogavage, once daily for two successive months. The activity of BACE1 was assayed by direct immunofluorescent method (DIF). The content of BACE1 protein was detected by Western blot.

RESULTSThe relative fluorescence units (RFU/microg) was 2.008 +/- 0.031 in the high dose PNS group, 2.221 +/- 0.029 in the low dose PNS group, and 2.267 +/- 0.076 in the huperzine A group, all lower than that in the normal control group (2.403 +/- 0.058; all P < 0.01). The content of BACE1 protein was 0.900 +/- 0.028 in the high dose PNS group, 1.000 +/- 0.032 in the low dose PNS group, and 0.837 +/- 0.080 in the huperzine A group, all lower than that in the normal control group (2.210 +/- 0.074, all P < 0.01).

CONCLUSIONPNS higher than 100 mg/kg could decrease the activity of BACE1 and down-regulate the content of BACE1 protein in the brain of SAMP8 mice.

Aging ; Alzheimer Disease ; metabolism ; Amyloid Precursor Protein Secretases ; metabolism ; Animals ; Aspartic Acid Endopeptidases ; metabolism ; Brain ; metabolism ; Disease Models, Animal ; Male ; Mice ; Panax notoginseng ; RNA, Messenger ; genetics ; Saponins ; pharmacology

OBJECTIVETo investigate the neuroprotective effect of estradiol on the release of excitatory amino acid (EAAs) mediator, and the activity of ATPase in cerebro-cortical synaptosome membrane of rats exposed to deltamethrin.

METHODSUsing HPLC to detect EAAs release, and colorimeter method to measure the activities of Na(+)-K(+)-ATPase, Mg(2+)-ATPase, Ca(2+)-ATPase, Ca(2+)-Mg(2+)-ATPase in the cerebro-cortical synaptosomes of ovariectomized rats exposed to deltamethrin (2 x 10(-5)mol/L), and treated with different doses of 17beta estradiol (10(-5), 10(-8), 10(-11) mol/L). Meanwhile, the estrogen receptor (ER) antagonist, tamoxifen, was used to investigate the effect on estradiol.

RESULTSThe release of Asp and Glu from the cerebro-cortical synaptosomes was significantly increased by 2 x 10(-5)mol/L deltamethrin exposure at the depolarizing state evoked by 50 mmol/L KCl, while 10(-8), 10(-11) mol/L 17beta estradiol could partly inhibit the effect of deltamethrin on the release of Asp (28.42%, 24.36%, respectively), Glu (21.52%, 14.57%, respectively). The activities of 4 kinds of ATPase were inhibited by 2 x 10(-4) mol/L deltamethrin, and these effects could be blocked by 10(-5) mol/L estradiol, while the activity of Ca(2+)-ATPase was increased by 10(-8), 10(-11) mol/L of estradiol. However, no obvious antagonistic effect of tamoxifen on the function of estradial on EAAs release or the activities of ATPase was found.

CONCLUSIONEstradiol showed certain neuroprotective effect on the release of EAAs and the inhibition on ATPase induced by deltamethrin. The effect of estradiol on synaptosomes may indicate the nongenetic mechanism of estradiol.

Animals ; Aspartic Acid ; secretion ; Cerebral Cortex ; drug effects ; metabolism ; Estradiol ; pharmacology ; Glutamic Acid ; secretion ; Insecticides ; toxicity ; Neurotoxicity Syndromes ; prevention & control ; Nitriles ; Pyrethrins ; toxicity ; Rats ; Rats, Wistar ; Synaptosomes ; drug effects ; metabolism

OBJECTIVETo investigate the effect of recombinant human hepatopoietin (rhHPO) and partial hepatectomy on rapidly induced expression of immediate early gene.

METHODSWe investigated the different gene expression within 1 hour after 2/3 partial hepatectomy by representational difference analysis and in primary cultured hepatocytes system.

RESULTSIn the expressed sequence tag (EST) library, we identified that most of these genes were immediate early gene, and found one new gene PC3 that might be associated to liver regeneration in the EST library. Moreover, PC3 gene was rapidly induced after 2/3 partial hepatectomy and the expressing peak was within 1~2 hours after operation. HPO can rapidly induce the expression of these genes (c-fos, LRF-1, and PC3, etc.) in primarily cultured rat hepatocyte, which might be one of HPO molecular mechanism on stimulating hepatocyte proliferation.

CONCLUSIONSrhHPO and partial hepatectomy can rapidly induce the expression of immediate early gene. PC3 gene is immediate early gene related to liver regeneration.

Animals ; Aspartic Acid Endopeptidases ; genetics ; Blotting, Northern ; Gene Expression Regulation ; drug effects ; Genes, Immediate-Early ; Hepatectomy ; Hepatocyte Growth Factor ; pharmacology ; Liver Regeneration ; genetics ; Proprotein Convertases ; RNA, Messenger ; biosynthesis ; Rats ; Rats, Wistar ; Recombinant Proteins ; pharmacology

OBJECTIVETo investigate the effects of exogenous hydrogen sulfide (H2S) on β-site APP cleaving enzyme 1 (BACE-1) and β-amyloid peptide (Aβ) metabolism in primary culture of neurons under high-glucose condition.

METHODSThe cortical neurons in primary culture under normal and high glucose (60 mmol/L) conditions for 24 h were exposed to 25, 50 and 100 µmol/L NaHS. Aβ1-42 concentration in the cell culture was measured by ELISA, and BACE-1 mRNA and protein levels were detected by fluorescent quantitative real-time PCR and Western blotting, respectively.

RESULTSCompared with the neurons cultured in normal glucose, the neurons exposed to high glucose showed significantly increased Aβ1-42 concentration and BACE-1 mRNA and protein expressions (P<0.05). Exposure to 25, 50 and 100 µmol/L NaHS significantly decreased Aβ1-42 concentration and BACE-1 mRNA and protein expressions in the high-glucose cell culture (P<0.05).

CONCLUSIONNeurons exposed to high glucose exhibit increased Aβ1-42 levels and BACE-1 mRNA and protein expressions, which can be concentration-dependently decreased by NaHS.

Amyloid Precursor Protein Secretases ; metabolism ; Amyloid beta-Peptides ; metabolism ; Animals ; Aspartic Acid Endopeptidases ; metabolism ; Cells, Cultured ; Culture Media ; chemistry ; Glucose ; chemistry ; Hydrogen Sulfide ; pharmacology ; Neurons ; drug effects ; metabolism ; Peptide Fragments ; metabolism ; Rats ; Rats, Sprague-Dawley

BACKGROUNDEpithelial-mesenchymal transition is a cellular process characterized by the loss of cell adhesion, inhibition of E-cadherin expression, and increased cell mobility. Cells without Napsin A are susceptible to transition. Further studies are required to investigate whether this transition can be reversed by restoration of Napsin A.

METHODSA Napsin A expression vector PLJM1-Napsin A plasmid was constructed and then transfected into the epithelial cell line A549 by lentivirus transfection to obtain A549-PLJM1-Napsin A cell line. Cell proliferation was assayed by 3-[4,5-dimethylthiazol-2-yl]-2,5 diphenyltetrazolium bromide and cell cycle was measured by flow cytometry. The E-cadherin, type I collagen, and focal adhesion kinase mRNA level was detected by reverse transcription-polymerase chain reaction. The Napsin A, E-cadherin, type I collagen, and focal adhesion kinase protein level in A549 cells was detected by Western blotting.

RESULTSTransforming growth factor-b1 induced epithelial-mesenchymal transition in A549 cells, as demonstrated by significant reduction of E-cadherin mRNA and protein levels (P < 0.01) as well as up-regulation of type I collagen (P < 0.01). Transfection of Napsin A in A549 cells can partially block the transforming growth factor-b1-regulated expression of E-cadherin and type I collagen (P < 0.01). In addition, transforming growth factor-b1-induced cell proliferation was inhibited by Napsin A (P < 0.01). Further study demonstrated that Napsin A caused G(0)/G(1) arrest and inhibited the expression of focal adhesion kinase (P < 0.01), a key protein in the integrin signaling pathway, in the in vitro epithelial-mesenchymal transition model.

CONCLUSIONSSustained Napsin A expression in A549 cells can inhibit the transforming growth factor-b1-induced epithelial-mesenchymal transition. This may be due to the Napsin A-mediated inhibition of focal adhesion kinase expression and integrin signaling pathway.

Aspartic Acid Endopeptidases ; genetics ; metabolism ; Cadherins ; genetics ; metabolism ; Cell Line ; Collagen Type I ; genetics ; metabolism ; Epithelial-Mesenchymal Transition ; drug effects ; genetics ; Focal Adhesion Protein-Tyrosine Kinases ; genetics ; metabolism ; Humans ; Transfection ; Transforming Growth Factor beta1 ; pharmacology