Metabolites Identification and Mechanism Prediction of Neobavaisoflavone In Vitro and In Vivo of Rats through UHPLC-Q-Exactive Plus Orbitrap MS Integrated Network Pharmacology

Neobavaisoflavone is an important isoflavone component isolated from Psoraleae Fructus. It is used extensively worldwide because of its antibacterial, antioxidant, anti-inflammatory, anticancer, and anti-osteoporotic activities. However, there is no systematic and comprehensive research on the metabolism of neobavaisoflavone in vivo and in vitro. The study aimed to analyze the metabolic characteristics and mechanism of neobavaisoflavone for the first time. Firstly, biological samples were pretreated by the solid-phase extraction (SPE) method, methanol precipitation, and acetonitrile precipitation. Secondly, the samples were analyzed on UHPLC-Q-Exactive Plus Orbitrap MS. Thirdly, metabolites were tentatively identified based on retention time, parallel reaction monitoring strategy, diagnostic product ions, and neutral loss fragments. A total of 72 metabolites of neobavaisoflavone were tentatively identified, including 28 in plasma, 43 in urine, 18 in feces, six in the liver, and four in the liver microsome. The results suggested that neobavaisoflavone mainly underwent glucuronidation, sulfation, hydroxylation, methylation, cyclization, hydration, and their composite reactions in vivo and in vitro. In addition, nine active components with high bioavailability and 191 corresponding targets were predicted by the Swiss Drug Design database. The 1806 items of GO and 183 KEGG signaling pathways were enriched. These results showed that metabolites expanded the potential effects of neobavaisoflavone. The present study would provide the scientific basis for the further exploitation and application of neobavaisoflavone.     

Development of a GC/Q-ToF-MS Method Coupled with Headspace Solid-Phase Microextraction to Evaluate the In Vitro Metabolism of β-Caryophyllene

Sample preparation remains both a challenging and time-consuming process in the field of bioanalytical chemistry. Many traditional techniques often require multi-step processes, which can introduce additional errors to the analytical method. Given the complexity of many biological matrices, thorough analyte extraction presents a major challenge to researchers. In the present study, a headspace solid-phase microextraction (HS-SPME) coupled with a GC/Q-ToF-MS method, was developed to quantify in vitro metabolism of β-caryophyllene by both human liver microsome (HLM) and S9 liver fractions. Validation of the method was demonstrated both in terms of linearity (R² = 0.9948) and sensitivity with a limit of detection of 3 ng/mL and a limit of quantitation of 10 ng/mL. In addition, the method also demonstrated both inter- and intra-day precision with the relative standard deviation (RSD) being less than 10% with four concentrations ranging from 50–500 ng/mL. Since this method requires no solvents and minimal sample preparation, it provides a rapid and economical alternative to traditional extraction techniques. The method also eliminates the need to remove salts or buffers, which are commonly present in biological matrices. Although this method was developed to quantify in vitro metabolism of one analyte, it could easily be adapted to detect or quantify numerous volatiles and/or semi-volatiles found in biological matrices.     

Crosstalk between Depression and Breast Cancer via Hepatic Epoxide Metabolism: A Central Comorbidity Mechanism

Breast cancer (BC) is a serious global challenge, and depression is one of the risk factors and comorbidities of BC. Recently, the research on the comorbidity of BC and depression has focused on the dysfunction of the hypothalamic–pituitary–adrenal axis and the persistent stimulation of the inflammatory response. However, the further mechanisms for comorbidity remain unclear. Epoxide metabolism has been shown to have a regulatory function in the comorbid mechanism with scattered reports. Hence, this article reviews the role of epoxide metabolism in depression and BC. The comprehensive review discloses the imbalance in epoxide metabolism and its downstream effect shared by BC and depression, including overexpression of inflammation, upregulation of toxic diols, and disturbed lipid metabolism. These downstream effects are mainly involved in the construction of the breast malignancy microenvironment through liver regulation. This finding provides new clues on the mechanism of BC and depression comorbidity, suggesting in particular a potential relationship between the liver and BC, and provides potential evidence of comorbidity for subsequent studies on the pathological mechanism.     

Chemical Synthesis and Biological Activities of Amaryllidaceae Alkaloid Norbelladine Derivatives and Precursors

Amaryllidaceae alkaloids (AAs) are a structurally diverse family of alkaloids recognized for their many therapeutic properties, such as antiviral, anti-cholinesterase, and anticancer properties. Norbelladine and its derivatives, whose biological properties are poorly studied, are key intermediates required for the biosynthesis of all ~650 reported AAs. To gain insight into their therapeutic potential, we synthesized a series of O-methylated norbelladine-type alkaloids and evaluated their cytotoxic effects on two types of cancer cell lines, their antiviral effects against the dengue virus (DENV) and the human immunodeficiency virus 1 (HIV-1), and their anti-Alzheimer’s disease (anti-cholinesterase and -prolyl oligopeptidase) properties. In monocytic leukemia cells, norcraugsodine was highly cytotoxic (CC₅₀ = 27.0 μM), while norbelladine was the most cytotoxic to hepatocarcinoma cells (CC₅₀ = 72.6 μM). HIV-1 infection was impaired only at cytotoxic concentrations of the compounds. The 3,4-dihydroxybenzaldehyde (selectivity index (SI) = 7.2), 3′,4′-O-dimethylnorbelladine (SI = 4.8), 4′-O-methylnorbelladine (SI > 4.9), 3′-O-methylnorbelladine (SI > 4.5), and norcraugsodine (SI = 3.2) reduced the number of DENV-infected cells with EC₅₀ values ranging from 24.1 to 44.9 μM. The O-methylation of norcraugsodine abolished its anti-DENV potential. Norbelladine and its O-methylated forms also displayed butyrylcholinesterase-inhibition properties (IC₅₀ values ranging from 26.1 to 91.6 μM). Altogether, the results provided hints of the structure–activity relationship of norbelladine-type alkaloids, which is important knowledge for the development of new inhibitors of DENV and butyrylcholinesterase.     

Electrochemical Studies on Effects of Psychostimulants,Opioids and Alcohol Intake Towards Level of Hydrogen Peroxide in the Brain Striatum

Hydrogen peroxide (H₂O₂) was the main types of many peroxides produced in living mammalian cells that consumed oxygen. In the brain, the main source of H₂O₂ was the superoxide dismutase (SOD)‐catalyzed reaction in mitochondria. However, the level of H₂O₂ would be elevated through administration of control drugs and alcohol by dopamine metabolism of monoamine oxidase. In this study, a H₂O₂ microsensor was used to investigate the level of H₂O₂ in the brain striatum after administration of methamphetamine (MAP), morphine (MrP) or ethanol (Eth). The placement of microsensor in the brain was done at coordinates A/P 1.1 from bregma, M/L+2.6 and D/V‐1.5. A working potential of +0.05 V vs. Ag/AgCl was applied. The H₂O₂ concentration was measured direct from the current generated by its catalytic reaction at the electro active surface of the electrode. A significant increase of H₂O₂ level was observed after 7 successive injections of the controlled drugs or alcohol. The initial measurement of H₂O₂ is essential as excess dosage of H₂O₂ during treatment will contribute to the formation of neurotoxin oxygenated radicals. The H₂O₂ was the precursor of O_2? and OH radicals. Thus, this study provided a mean to monitor H₂O₂ level in the brain.     

Langendorff灌流心脏的31P NMR谱测定

测定了Langendorff灌流大鼠和兔心脏的³¹P NMR谱.观测到PCr、ATP、SP、P_i以及PME和GPC等含磷代谢物的共振峰,各谱峰之间实现了很好的分辨.在3~5min的累加时间内测得的图谱具有较好的S/N,可用于以上含磷代谢物的定量测定,以高能含磷化合物ATP和PCr相对峰强度以及冠脉流量的变化对灌流大鼠心脏代谢的稳定性进行了考察,结果表明,在本实验条件下大鼠心脏的能量储存至少可稳定2h,用所建方法以3min的时间间隔对同一大鼠心脏缺血21min及再灌注12min过程中心脏的³¹P NMR谱进行了连续跟踪测定,并初步观测了缺血及再灌注过程中心肌细胞内ATP、PCr与P_i之间的消长关系.     

Metabolism of xanthohumol and isoxanthohumol, prenylated flavonoids from hops (Humulus lupulus L.), by human liver microsomes

The female flowers of hops (Humulus lupulus L.) used to flavor beer contain the prenylated flavonoids xanthohumol (XN) and isoxanthohumol (IX). IX is moderately estrogenic in vitro and XN has pharmacological properties that might make it useful as a cancer chemopreventive agent. The metabolism of these dietary flavonoids was investigated in vitro using human liver microsomes. Hydroxylation of a prenyl methyl group was the primary route of oxidative metabolism forming either cis or trans hydroxylated metabolites of IX but only the trans isomer of XN. The double bond on the prenyl group of both compounds formed an epoxide which was opened by an intramolecular reaction with the neighboring hydroxyl group. The potent phytoestrogen 8-prenylnaringenin (8-PN) was detected as a demethylation product of IX. However, the analogous demethylation reaction was not observed for XN. Since XN can be converted to IX through acid-catalyzed cyclization in the stomach, XN might contribute to the in vivo levels of estrogenic 8-PN following consumption of hops extracts.     

Microcalorimetric study of virus infection; The effects of hyperthermia and 1b recombinant homo interferon on the infection process of BHK-21 cells by foot and mouth disease virus

The metabolic process of BHK-21 cell line infected by foot and mouth disease virus (FMDV) was determined by using LKB-2277 Bioactivity Monitor. The aim of the present study is to investigate the metabolic thermal power of the virus infection process of BHK-21 cells, the effects of combinational treatments of hyperthermia and 1b recombinant homo interferon on this process. In contrast to the metabolic thermal power of uninfected BHK-21 cells, the thermogenetic curves show that the energy metabolism mechanisms of BHK-21 cells were significantly changed by the virus infection process. The maximum thermal power decreased and the time needed to reach the maximal thermal power increased with the increasing concentration of interferon. The results also show that the infection process was thermosensitive. But no apparent synergetic effect of the combinational treatments of hyperthermia and interferon was observed. The present microcalorimetric results are in accordance with the cytomorphology observations.     

Reminiscence of 40-year research on nitrogen metabolism

This article summarizes my research over 40 years. The main theme of my work is nitrogen metabolism of amino acids, though later I focused on protein turnover in the cell. In the first years of my research work, I was busy dissecting the pathways involved in the metabolism of certain amino acids and their related enzymes. Then I became interested in the physiology and regulation of matabolism of these amino acids. For that, I used primary cultured hepatocytes, which contain many liver-specific enzymes. However, this play field was very rough around 1970 and hence I had to smooth them (differentiated) first. We discovered a specific growth factor (hepatocyte growth factor, HGF) in rat platelets. Exceptionally, I also worked on branched chain amino acids (valine, leucine and isoleucine). These amino acids are not efficiently metabolized in the liver, so I had to consider the physiology of extrahepatic tissues as well. Finally, I came across a huge protease complex, the proteasome. Whether these players, small amino acid metabolizing enzymes and the huge protease complex, danced well in harmony on my playground or not, I still do not know.