Identification and Simultaneous Determination of the Main Toxical Pyrrolizidine Alkaloids in a Compound Prescription of Traditional Chinese Medicine: Qianbai Biyan Tablet

Qianbai biyan tablet (QT) is a compound prescription of traditional Chinese medicine which is used to treat nasal congestion, rhinitis, and nasosinusitis, with Senecio scandens as its main plant material. Several pyrrolizidine alkaloids (PAs) were reported in Senecio scandens and others of Senecio species. Although Senecio scandens is assigned as the legal plant material of QT, whether replaced use of it by other Senecio plants can bring toxicity is unknown because of the lack of quantitative data about toxic PAs between different Senecio species. In the present study, adonifoline, senkirkine, and another PA presumed as emiline have been identified in QT; however, there was no senecionine detected in all tablets. PA contents in QTs varied in different companies and different batches. Adonifoline existed only in Senecio scandens, and senecionine was detected in all eight Senecio plants investigated in the present study. Data showed that replaced use of Senecio scandens with a low level of senecionine by other Senecio plants such as Senecio vulgaris containing a high level of senecionine is advertised to be forbidden. Data of the present study may be used as a reference to make new drug quality regularity and recommendation guideline for the safety of QT

Fatty acid metabolites of Dendrobium nobile were positively correlated with representative endophytic fungi at altitude

IntroductionAltitude, as a comprehensive ecological factor, regulates the growth and development of plants and microbial distribution. Dendrobium nobile (D. nobile) planted in habitats at different elevations in Chishui city, also shows metabolic differences and endophytes diversity. What is the triangular relationship between altitude, endophytes, and metabolites?MethodsIn this study, the diversity and species of endophytic fungi were tested by ITS sequencing and metabolic differences in plants were tested by UPLC–ESI–MS/MS. Elevation regulated the colonization of plant endophytic fungal species and fatty acid metabolites in D. nobile.ResultsThe results indicate that and high altitude was better for the accumulation of fatty acid metabolites. Therefore, the high-altitude characteristic endophytic floras were screened, and the correlation with fatty acid metabolites of plants was built. The colonization of T. rubrigenum, P. Incertae sedis unclassified, Phoma. cf. nebulosa JZG 2008 and Basidiomycota unclassified showed a significantly positive correlation with fatty acid metabolites, especially 18-carbon-chain fatty acids, such as (6Z,9Z,12Z)-octadeca-6,9,12-trienoic acid, 3,7,11,15-tetramethyl-12-oxohexadeca-2,4-dienoic acid and Octadec-9-en-12-ynoic acid. What is more fascinating is these fatty acids are the essential substrates of plant hormones.DiscussionConsequently, it was speculated that the D. nobile- colonizing endophytic fungi stimulated or upregulated the synthesis of fatty acid metabolites and even some plant hormones, thus affecting the metabolism and development of D. nobile

Apelin-13 Administration Protects Against LPS-Induced Acute Lung Injury by Inhibiting NF-κB Pathway and NLRP3 Inflammasome Activation

Background/Aims: Acute lung injury (ALI) is induced by a variety of external and internal factors and leads to acute progressive respiratory failure. Previous studies have shown that apelin-13 can decrease the acute lung injury induced by LPS, but the specific mechanism is unclear. Therefore, a mouse lung injury model and a cell model were designed to explore the mechanism of how apelin-13 alleviates the acute lung injury caused by LPS. Methods: The effect of apelin-13 on LPS-induced structural damage was determined by H&E staining and by the wet/dry weight ratio. The related inflammatory factors in BALF were examined by ELISA. The apoptotic pathway and the NF-κB and NLRP3 inflammasome pathways were evaluated by using Western blotting and immunofluorescence staining. Results: LPS induced the structural damage and the production of inflammatory cytokines in the lung tissues of mice. These deleterious effects were attenuated by apelin-13 administration. The protective effects of apelin-13 were associated with decreased reactive oxygen species (ROS) formation and the inhibition of the activation of the NF-κB and NLRP3 inflammasome pathways in mice and in Raw264.7 cells. Conclusion: Taken together, these data suggest that apelin-13 administration ameliorates LPS-induced acute lung injury by suppressing ROS formation, as well as by inhibiting the NF-κB pathway and the activation of the NLRP3 inflammasome in the lungs

The role of organic anion transport protein 1a4 in drug delivery and diseases: a review

Abstract OATP1A4 is an important member of the family of organic anion transporting polypeptides (OATPs), which is generally thought to mediate cellular uptake of endogenous and exogenous substances, such as bile acids, drugs and environmental toxins. Recent studies have found that Oatp1a4 plays an important role in drug passage through the blood-brain barrier and is expected to be an important target for drugs used to treat central diseases. Oatp1a4 has also been associated with various diseases such as cholestasis. differences in Oatp1a4 across age and sex have also become an area of concern for precision drug administration. Therefore, this paper presents a systematic review of Oatp1a4 expression in drug transport and various physiopathological states

Rosiglitazone induces hepatocyte injury by increasing DCA accumulation through OATP1A4 inhibiting in mice

Rosiglitazone (RL), a second-generation thiazolidinedione (TZD), is an oral antidiabetic agent that is reportedly associated with hepatotoxicity. However, the molecular mechanism of Rosiglitazone induced hepatotoxicity has not been reported yet. To minimize clinical risk, in the present study, toxicity was evaluated initially in C57BL/6 mice to confirm the hepatotoxicity of Rosiglitazone. Subsequently, the transcription of genes, the up-regulated expression of proteins and changes of metabolite spectrum in bile acid system in accordance with system pharmacology and multi-omics profiling strategy were investigated to reveal its toxic mechanism. Results show an obvious Rosiglitazone induced hepatocyte injury in our experiment. Changes in bile acid profiles revealed that Rosiglitazone administration increased the concentration of deoxycholic acid (DCA) in the liver, but decreased those of other bile acids. Transcriptomic and proteomic data showed that while the expression of efflux transporters ABCC3, ABCC4 and ABCB11 was upregulated following Rosiglitazone treatment, the expression of two hepatic uptake transporters, OATP1A1 and OATP1A4, which were negatively associated with DCA accumulation, were significantly downregulated. These results indicated that cholestasis, especially the accumulation of DCA in the liver, is the primary factors responsible for Rosiglitazone induced hepatocyte injury. The mechanism of Rosiglitazone induced DCA accumulation in the liver may be the result of stimulated intestinal permeability and increased biosynthesis and uptake of DCA and weakening of DCA amidation to reduce DCA efflux due to the inhibition of OATP1A1 and OATP1A4 expression in the liver

Differential effects of medium- and long-term high-fat diets on the expression of genes or proteins related to nonalcoholic fatty liver disease in mice

Abstract Non-alcoholic fatty liver disease (NAFLD) is now considered to be the most common liver disease worldwide, caused by fat deposition in hepatocytes. High-fat diet is considered to be a major lifestyle factor predisposing to NAFLD. However, the effect of different cycles of high-fat diets on changes in NAFLD-related gene and protein expression is unclear. In this study, NAFLD mouse models were established by feeding C57BL/6 male mice a high-fat diet for 16 and 38 weeks. The transcriptome and proteome of mouse liver were profiled by RNA sequencing and high-resolution mass spectrometry, respectively. The results show that accumulation of liver lipids was observed at 38 weeks of treatment on a high-fat diet. At the same time, the expression profiles of 1329 genes and 802 proteins involved in NAFLD were changed, with a total of 234 genes and 37 proteins significantly changing by more than twice. These differentially expressed genes and proteins were significantly enriched in fatty acid metabolism and organic acid biosynthesis. 18 genes and their corresponding protein overlaps were identified using Venn diagrams, and most of them were regulated by high-fat diet in an aging-dependent manner. All in all, our study is valuable for understanding the high-fat diet on the developmental process of NAFLD

Construction of in vitro liver-on-a-chip models and application progress

Abstract The liver is the largest internal organ of the human body. It has a complex structure and function and plays a vital role in drug metabolism. In recent decades, extensive research has aimed to develop in vitro models that can simulate liver function to demonstrate changes in the physiological and pathological environment of the liver. Animal models and in vitro cell models are common, but the data obtained from animal models lack relevance when applied to humans, while cell models have limited predictive ability for metabolism and toxicity in humans. Recent advancements in tissue engineering, biomaterials, chip technology, and 3D bioprinting have provided opportunities for further research in in vitro models. Among them, liver-on-a-Chip (LOC) technology has made significant achievements in reproducing the in vivo behavior, physiological microenvironment, and metabolism of cells and organs. In this review, we discuss the development of LOC and its research progress in liver diseases, hepatotoxicity tests, and drug screening, as well as chip combinations. First, we review the structure and the physiological function of the liver. Then, we introduce the LOC technology, including general concepts, preparation materials, and methods. Finally, we review the application of LOC in disease modeling, hepatotoxicity tests, drug screening, and chip combinations, as well as the future challenges and directions of LOC

Application of single-cell RNA sequencing technology in liver diseases: a narrative review.

Objective: This review aimed to summarize the application of single-cell transcriptome sequencing technology in liver diseases. Background: The increasing application of single-cell ribonucleic acid (RNA) sequencing (scRNA-seq) in life science and biomedical research has greatly improved our understanding of cellular heterogeneity in immunology, oncology, and developmental biology. scRNA-seq has proven to be a powerful tool for identifying and classifying cell subsets, characterizing rare or small cell subsets and tracking cell differentiation along the dynamic cell stages. Globally, liver disease has high rates of morbidity and mortality, and its exact pathological mechanism remains unclear, current treatment options are limited to clearance of the underlying cause or liver transplantation, which cannot overwhelm and cure liver diseases. scRNA-seq provides many novel insights for healthy and diseased livers. Methods: In this review, we searched for related articles in the PubMed database and summarized the advances of scRNA-seq in revealing the molecular mechanisms of liver development, regeneration, and disease. We also discussed the challenges and future application potential of scRNA-seq, which is expected to enhance the ability to explore the field of liver research and accelerate the clinical application of liver precision medicine. Conclusions: With the continuous improvement of scRNA-seq technology, scRNA-seq is expected to unlock new avenues for liver biology exploration, liver disease diagnosis, and personalized treatment, which will pave the way for breakthrough innovation in personalized medicine

Establishment of fingerprints and determination of various ingredients of yanlishuang pills by GC-MS

Abstract Yanlishuang Pills is a kind of traditional Chinese medicine used to treat pharyngitis widely. In this study, we used gas chromatography tandem mass spectrometry (GC-MS) to establish a method for the fingerprint and quantitative analysis of the four major components of Yanlishuang Pills, which can provide a more reliable method for its quality control. We used the software “Chromatographic Fingerprint Similarity Evaluation System for Traditional Chinese Medicine”, version A, 2004, to obtain fingerprint using the averaging method with a time width of 0.1. The peak with the largest peak area was used as the reference peak to determine the shared peaks and generate the common pattern. Then the main components of the Yanlishuang Pills were identified and their contents were determined in GC-MS SIM mode using internal standard method.The fingerprint established by GC-MS were reproducible, and a total of 18 common peaks were identified in the fingerprint of 13 batches of samples, and the similarity of the fingerprint of each batch of samples was above 0.99. The concentrations of camphor, menthone, borneol and menthol of the four main ingredients of the Yanlishuang Pills were linearly well within the range of 25.13-150.78 μg/mL (r = 0.9995), 28.77-172.62 μg/mL (r = 0.9991), 299.70-1798.20 μg/mL (r = 0.9997), 121.98-731.88 μg/mL (r = 0.9997), and the average recoveries were 102.02% (RSD of 1.3%), 96.10% (RSD of 1.0%), 102.71% (RSD of 1.3%), 102.58% (RSD of 1.1%), respectively, with good precision, reproducibility, and stability within 16 h. The camphor content of the 13 batches of samples was 5.6025-8.3662 mg/g, menthone content was 4.7871-5.8936 mg/g, borneol content was 88.0034-133.0969 mg/g and menthol was 40.2017-61.9466 mg/g. The fingerprints of the Yanlishuang Pills established by GC-MS were characterized by a common pattern, and the simultaneous determination of camphor, menthone, borneol and menthol in the Yanlishuang Pills was rapid, simple and accurate. In conclusion, the determination of the content of multiple ingredients combined with fingerprinting can provide a more comprehensive control of the quality of Yanlishuang Pills

Identification of Human UDP-Glucuronosyltransferase Involved in Gypensapogenin C Glucuronidation and Species Differences

Gypensapogenin C (GPC) is one of the important aglycones of Gynostemma pentaphyllum (GP), which is structurally glucuronidated and is highly likely to bind to UGT enzymes in vivo. Due to the important role of glucuronidation in the metabolism of GPC, the UDP-glucuronosyltransferase metabolic pathway of GPC in human and other species’ liver microsomes is investigated in this study. In the present study, metabolites were detected using high-performance liquid chromatography–tandem mass spectrometry (LC–MS/MS). The results show that GPC could generate a metabolite through glucuronidation in the human liver microsomes (HLMs). Additionally, chemical inhibitors combined with recombinant human UGT enzymes clarified that UGT1A4 is the primary metabolic enzyme for GPC glucuronidation in HLMs according to the kinetic analysis of the enzyme. Metabolic differential analysis in seven other species indicated that rats exhibited the most similar metabolic rate to that of humans. In conclusion, UGT1A4 is a major enzyme responsible for the glucuronidation of GPC in HLMs, and rats may be an appropriate animal model to evaluate the GPC metabolism