Host-defence caerin 1.1 and 1.9 peptides suppress glioblastoma U87 and U118 cell proliferation through the modulation of mitochondrial respiration and induce the downregulation of CHI3L1.

Glioblastoma, the most aggressive form of brain cancer, poses a significant global health challenge with a considerable mortality rate. With the predicted increase in glioblastoma incidence, there is an urgent need for more effective treatment strategies. In this study, we explore the potential of caerin 1.1 and 1.9, host defence peptides derived from an Australian tree frog, in inhibiting glioblastoma U87 and U118 cell growth. Our findings demonstrate the inhibitory impact of caerin 1.1 and 1.9 on cell growth through CCK8 assays. Additionally, these peptides effectively curtail the migration of glioblastoma cells in a cell scratch assay, exhibiting varying inhibitory effects among different cell lines. Notably, the peptides hinder the G0/S phase replication in both U87 and U118 cells, pointing to their impact on the cell cycle. Furthermore, caerin 1.1 and 1.9 show the ability to enter the cytoplasm of glioblastoma cells, influencing the morphology of mitochondria. Proteomics experiments reveal intriguing insights, with a decrease in CHI3L1 expression and an increase in PZP and JUNB expression after peptide treatment. These proteins play roles in cell energy metabolism and inflammatory response, suggesting a multifaceted impact on glioblastoma cells. In conclusion, our study underscores the substantial anticancer potential of caerin 1.1 and 1.9 against glioblastoma cells. These findings propose the peptides as promising candidates for further exploration in the realm of glioblastoma management, offering new avenues for developing effective treatment strategies.

Effects of three Huanglian-derived polysaccharides on the gut microbiome and fecal metabolome of high-fat diet/streptozocin-induced type 2 diabetes mice.

Plant-derived polysaccharides are important components for biological functions. The objective of this study is to study the mechanisms by which polysaccharides from three Huanglian (Rhizome Coptidis, HL) of Coptis chinensis, C. deltoidea, and Coptis teeta affect type 2 diabetes mellitus (T2DM) by analyzing the gut microbiome and their metabolites. A long-term high-fat diet (HFD) combined with streptozocin (STZ) induction was used to construct the T2DM mice model. The histopathology of liver, pancreas, and colon, biochemical indexes related to mice were determined to assess the ameliorative effects of these three HL polysaccharides (HLPs) on T2DM. The results indicated that oral HLPs improved hyperglycemia, insulin resistance, blood lipid levels, and ß-cell function. Further, HLPs elevated the growth of advantageous beneficial bacteria within the gut microbiota and raised the concentrations of short-chain fatty acids (SCFAs), particularly butyric acid. Metabolic analyses showed that HLPs ameliorated the effects of T2DM on microbial-derived metabolites and related metabolic pathways, especially the biosynthetic pathways of phenylalanine, tyrosine, and tryptophan. In the combined analysis, many associations of T2DM-related biochemical indicators with gut microbes and their metabolites were extracted, which suggested the important role of gut microbiome and fecal metabolome in the amelioration of type 2 diabetes mellitus by HLPs.

Microbiome-Metabolome Analysis Insight into the Effects of the Extract of Phyllanthus emblica L. on High-Fat Diet-Induced Hyperlipidemia.

The fruit of Phyllanthus emblica L. (FEPE) has a long history of use in Asian folk medicine. The main bioactive compounds in FEPE are polyphenols, known for their potent antioxidant, anti-inflammatory, and hypolipidemic activities. The present study aimed to investigate the intervention effect of FEPE (100 and 200 mg/kg) on hyperlipidemia for 8 weeks and preliminarily explored the potential mechanism by microbiome-metabolome analysis. The results showed that a high-dose FEPE (200 mg/kg) effectively alleviated dyslipidaemic symptoms and body weight gain in hyperlipidemic mice induced by a high-fat diet (HFD). Microbiome analysis showed that FEPE altered the structure of the intestinal microbiota, which included an increase in specific probiotics (such as Akkermansia, Anaerovorax, and Bacteroides) and a decrease in harmful bacteria (including A2, Acetitomaculum, Candidatus_Arthromitus, Lachnospiraceae_NK4A136_group, Lachnospiraceae_NK4B4_group, Rikenella, and Streptococcus), as well as a reduction in the level of short-chain fatty acids (SCFAs). In addition, significant changes in the hepatic metabolome were observed, and eight key metabolites associated with betaine metabolism, lysine degradation, methionine metabolism, and fatty acid metabolism pathways were primarily filtered. The correlated analysis identified several key "microbiota-metabolite" axes in the treatment of hyperlipidemia by FEPE extract. In conclusion, the present study is expected to provide a basis for treating hyperlipidemia with FEPE from the perspective of the microbiome-liver metabolome axis.

Relatively high light inhibits reserves degradation in the Coptis chinensis rhizome during the leaf expansion by changing the source-sink relationship.

The early spring is a seasonal high-light "window" for new leaf growth and photosynthetic carbon capture by the shade-tolerant evergreen understory plants. However, it remains unclear how light regulates the source-sink relationship between rhizome (RO), mature leaf (ML), and immature leaf (IL) during Coptis chinensis leaf expansion. Understanding this relationship is essential to reducing RO reserve degradation and ultimately promote RO biomass accumulation. The plants grew in an artificial climate chamber with low (50 µmol m-2 s-1) and relatively high (200 µmol m-2 s-1) light intensity treatments. Leaf fluorescence, foliar phosphorus (P) fractions, soluble sugars, starch, total P, and alkaloid concentrations in ILs, MLs, and RO were measured, and 13C labeling was used to indicate the direction of photosynthetic carbon flow between organs. The plants grown under high light intensity had higher levels of starch in RO and higher RO biomass at the end of the year compared to those grown under low light intensity. The photosystem II (PSII) operating efficiency [Y(II)], relative electron transport rate (rETR), and photochemical quenching (qP), as well as sucrose and glucose, in ILs and MLs under relatively high light, was higher than those under low light. The glucose and starch concentrations in ILs at 35 d was significantly higher than that at 15 d when plants were under 200 µmol m-2 s-1, while they were not significantly changed and remained low at 50 µmol m-2 s-1. The 13C was detected in the RO when plants were grown at 200 µmol m-2 s-1, regardless of ILs and MLs 13C labeling, while no 13C was detected in the RO when plants were under 50 µmol m-2 s-1. Additionally, the proportion of photosynthetic transport from ILs to MLs was significantly higher than that from MLs to ILs under the 50-µmol m-2 s-1 limit. Total P concentration in ILs was lower under relatively high light, but there was no difference in nucleic acid P concentration in ILs under the two light intensity treatments. The alkaloid concentration in RO was lower under 200 µmol m-2 s-1 than that under 50 µmol m-2 s-1. We propose that relatively high light reduces the need for carbohydrates and P stored in the RO to support IL growth by (1) accelerating the sink-to-source transition in ILs, which inhibits the use of reserves in the RO; (2) using energy from MLs to support IL growth, thereby reducing RO reserve consumption, and (3) reducing the demand for P by investing less in the development of photosynthetic machinery. Furthermore, under low light, MLs serve as a sink and rely on other organs for support, directly or indirectly exacerbating the reserves lost in the RO.

Comprehensive analysis of the complete mitochondrial genomes of three Coptis species (C. chinensis, C. deltoidea and C. omeiensis): the important medicinal plants in China.

Coptis plants (Ranunculaceae) contain high levels of isoquinoline alkaloids and have a long history of medicinal use. Coptis species are of great value in pharmaceutical industries and scientific research. Mitochondria are considered as one of the central units for receiving stress signals and arranging immediate responses. Comprehensive characterizations of plant mitogenomes are imperative for revealing the relationship between mitochondria, elucidating biological functions of mitochondria and understanding the environmental adaptation mechanisms of plants. Here, the mitochondrial genomes of C. chinensis, C. deltoidea and C. omeiensis were assembled through the Nanopore and Illumina sequencing platform for the first time. The genome organization, gene number, RNA editing sites, repeat sequences, gene migration from chloroplast to mitochondria were compared. The mitogenomes of C. chinensis, C. deltoidea and C. omeiensis have six, two, two circular-mapping molecules with the total length of 1,425,403 bp, 1,520,338 bp and 1,152,812 bp, respectively. The complete mitogenomes harbors 68-86 predicted functional genes including 39-51 PCGs, 26-35 tRNAs and 2-5 rRNAs. C. deltoidea mitogenome host the most abundant repeat sequences, while C. chinensis mitogenome has the largest number of transferred fragments from its chloroplasts. The large repeat sequences and foreign sequences in the mitochondrial genomes of Coptis species were related to substantial rearrangements, changes in relative position of genes and multiple copy genes. Further comparative analysis illustrated that the PCGs under selected pressure in mitochondrial genomes of the three Coptis species mainly belong to the mitochondrial complex I (NADH dehydrogenase). Heat stress adversely affected the mitochondrial complex I and V, antioxidant enzyme system, ROS accumulation and ATP production of the three Coptis species. The activation of antioxidant enzymes, increase of T-AOC and maintenance of low ROS accumulation in C. chinensis under heat stress were suggested as the factors for its thermal acclimation and normal growth at lower altitudes. This study provides comprehensive information on the Coptis mitogenomes and is of great importance to elucidate the mitochondrial functions, understand the different thermal acclimation mechanisms of Coptis plants, and breed heat-tolerant varieties.

Characterization of the anti-AChE potential and alkaloids in Rhizoma Coptidis from different Coptis species combined with spectrum-effect relationship and molecular docking.

Coptis species are the main source of Rhizoma Coptidis (RC) drugs, which have always been used to treat Alzheimer's disease in the clinical experience of ancient China. However, many species of this genus have been largely underutilized until now. With this fact, this research has been designed to investigate for the first time the anti-acetylcholinesterase (AChE) property of different extracts for RC drugs from four Coptis species (C. chinensis, C. deltoidea, C. teeta and C. omeiensis) and to quantify the main alkaloids. Petroleum ether, ethyl acetate and n-butanol fractions of RC drugs were sequentially collected using an accelerated solvent extraction technique. Spectrum-effect relationship and molecular docking were applied to analyse the relationships between alkaloids and AChE inhibitory activity. The N-butanol extract was proven to be the main active fraction, and C. teeta may be the best source of RC drugs for Alzheimer's disease treatment, with significantly lower IC 20, IC 50 and IC 80 values for AChE inhibition. The UPLC/QqQ-MS quantitative analysis showed that the accumulations of 10 alkaloids in RC drugs from different sources greatly varied. Three data processing methods (Random forest, Boruta and Pearson correlation) comprehensively analysed the spectrum-effect relationship and revealed that columbamine, berberine and palmatine were the most important AChE inhibitors that could be used as quality markers to select RC drugs for Alzheimer's disease treatment. In addition, the dominant compounds were successfully docked against AChE to verify the binding affinity and interactions with the active site. The present study can contribute to the reasonable development and utilization of RC drugs from different sources, especially to provide certain evidence for their application in the treatment of Alzheimer's disease.

An integrated chemical characterization based on FT-NIR, GC-MS and LC-MS for the comparative metabolite profiling of wild and cultivated agarwood.

Agarwood is a well-known and precious traditional Chinese medicine, has been widely applied as drugs and spices for century. The large demand for this material has deeply stimulated the emergence of numerous cultivated products. However, it is difficult to distinguish wild agarwood from cultivated agarwood, and the chemical composition difference between them is not clear. In this study, an integrated method of Fourier transform near-infrared (FT-NIR), gas chromatography-mass spectrometry (GC-MS) and ultraperformance liquid chromatography Quadrupole-Exactive Orbitrap tandem mass spectrometry (UHPLC-Q-Exactive Orbitrap/MS) was developed to explore chemical variation between wild and cultivated agarwood in combination with multivariate statistical analysis. Twenty-four wild and cultivated agarwood samples were collected from different regions. FT-NIR profiles were used to obtain the holistic metabolic characterization in combination with principal component analysis (PCA). A total of seventy-six and seventy-nine metabolites, including volatile components and 2-(2-phenethyl) chromones derivatives, were successfully identified by GC-MS and UHPLC-Q-Exactive Orbitrap/MS, respectively. Thereafter, the orthogonal-partial least square method-discriminant analysis (OPLS-DA) and variable importance in the projection (VIP) were used to screen potential characteristic chemical components (VIP > 1) in wild and cultivated agarwood, respectively. Finally, eight key chemical markers were putatively identified by two techniques to distinguish agarwood from different origins, which can be found that sesquiterpenes, aromatics, terpenoids, 2-(2-phenylethyl) chromones of the flidersia type (FTPECs) and tetrahydro-2-(2-phenylethyl) chromones (THPECs) are the most important metabolites. Summary, this research presented a comprehensive metabolomic variation between wild and cultivated agarwood on the basis of a multi-technology platform, which laid a foundation for distinguishing the two ecotypes of agarwood and was conducive to the quality control of this resource.

Jatrorrhizine: A Review of Sources, Pharmacology, Pharmacokinetics and Toxicity.

Jatrorrhizine, an isoquinoline alkaloid, is a bioactive metabolite in common medicinal plants, such as Berberis vernae Schneid., Tinospora sagittata (Oliv.) Gagnep. and Coptis chinensis Franch. These plants have been used for centuries in traditional medicine for their wide-ranging pharmacological properties. This review emphasizes the latest and comprehensive information on the sources, pharmacology, pharmacokinetics and toxicity of jatrorrhizine. Studies on this alkaloid were collected from scientific internet databases, including the Web of Science, PubMed, ScienceDirect, Google Scholar, Elsevier, Springer, Wiley Online Library and Europe PMC and CNKI, using a combination of keywords involving "jatrorrhizine", "sources", "pharmacology," "pharmacokinetics," and "toxicology". Jatrorrhizine exhibits anti-diabetic, antimicrobial, antiprotozoal, anticancer, anti-obesity and hypolipidemic properties, along with central nervous system activities and other beneficial activity. Studies of jatrorrhizine have laid the foundation for its application to the treatment of various diseases, but some issues still exist. Further investigations might emphasize 1) specific curative mechanisms of jatrorrhizine and clinical utility, 2) application prospect in the treatment of metabolic disorders, 3) comprehensive investigations of the toxicity mechanisms and 4) interactions of jatrorrhizine with other pharmaceuticals and development of derivatives.

Integrated metabolomics and ligand fishing approaches to screen the hypoglycemic ingredients from four Coptis medicines.

Rhizoma Coptidis, which is mainly originated from the rhizomes of Coptis chinensis, C. deltoidea, C. omeiensis and C. teeta, has been proved to possess a superior anti-diabetic effect in clinic. However, the metabolic characterization and the hypoglycemic ingredients among these Coptis species remain unclear. In this study, we employed an integrated strategy to screen the bioactive ingredients based on metabolomics and ligand fishing approaches. First, the ultra high-performance liquid chromatography coupled to quadruple time-of-flight tandem mass spectrometry (UHPLC-Q-TOF-MS/MS) was used for qualitative identification of four Coptis rhizomes. After prescreening by α-glucosidase inhibition assay, an affinity ultrafiltration system was constructed to fish out hypoglycemic ingredients from the fractions with superior activity, and verified by molecular docking on a virtual platform. The distribution of major compounds suggested the four Coptis rhizomes possess similar metabolic profiles, mainly including alkaloids and phenylpropanoids. Besides, eight compounds (magnoflorine, groenlandicine, jatrorrhizine, epiberberine, columbamine, coptisine, palmatine and berberine) from the n-butanol fraction were specifically bound to α-glucosidase, and considered as hypoglycemic ingredients of Rhizoma Coptidis. Molecular docking revealed that the inhibitors bound to α-glucosidase mainly by hydrophobic interaction, hydrogen bond interaction and π-π interaction. Summary, this research leads a more systematic and comprehensive study on metabolic characterization and hypoglycemic ingredients of Rhizoma Coptidis, which can provide a theoretical basis for the further clinical application.

A Systematic Review on Traditional Uses, Sources, Phytochemistry, Pharmacology, Pharmacokinetics, and Toxicity of Fritillariae Cirrhosae Bulbus.

Fritillariae Cirrhosae Bulbus (known as chuanbeimu in Chinese, FCB) is a famous folk medicine which has been widely used to relieve cough and eliminate phlegm for thousands of years in China. The medicine originates from dried bulbs of six species of Fritillaria which are distributed in the temperate zone of the Northern Hemisphere. Increasing attention has been paid to FCB because of its excellent medicinal value such as being antitussive, expectorant, analgesic, anticancer, anti-inflammatory, and antioxidative. During the past years, a large number of research studies have been conducted to investigate the phytochemistry, pharmacology, and pharmacokinetics of FCB. A range of compounds have been isolated and identified from FCB, including alkaloids, saponins, nucleosides, organic acids, terpenoids, and sterols. Among them, alkaloids as the main active ingredient have been illustrated to exert significant therapeutic effects on many diseases such as cancer, acute lung injury, chronic obstructive pulmonary disease, asthma, Parkinson's disease, and diabetes. Due to the excellent medical value and low toxicity, FCB has a huge market all over the world and triggers a growing enthusiasm among researchers. However, there is still a lack of systematic review. Hence, in this work, we reviewed the FCB-based articles published in Sci Finder, Web of Science, PubMed, Google Scholar, CNKI, and other databases in the recent years. The traditional uses, sources, phytochemistry, pharmacology, pharmacokinetics, and toxicity of FCB were discussed in the review, which aims to provide a reference for further development and utilization of FCB.

An integrated spectroscopic strategy to trace the geographical origins of emblic medicines: Application for the quality assessment of natural medicines.

Emblic medicine is a popular natural source in the world due to its outstanding healthcare and therapeutic functions. Our preliminary results indicated that the quality of emblic medicines might have an apparent regional variation. A rapid and effective geographical traceability system has not been designed yet. To trace the geographical origins so that their quality can be controlled, an integrated spectroscopic strategy including spectral pretreatment, outlier diagnosis, feature selection, data fusion, and machine learning algorithm was proposed. A featured data matrix (245 × 220) was successfully generated, and a carefully adjusted RF machine learning algorithm was utilized to develop the geographical traceability model. The results demonstrate that the proposed strategy is effective and can be generalized. Sensitivity (SEN), specificity (SPE) and accuracy (ACC) of 97.65%, 99.85% and 97.63% for the calibrated set, as well as 100.00% predictive efficiency, were obtained using this spectroscopic analysis strategy. Our study has created an integrated analysis process for multiple spectral data, which can achieve a rapid, nondestructive and green quality detection for emblic medicines originating from seventeen geographical origins.

Full-length transcriptome analysis of Coptis deltoidea and identification of putative genes involved in benzylisoquinoline alkaloids biosynthesis based on combined sequencing platforms.

KEY MESSAGE: The study carry out comprehensive transcriptome analysis of C. deltoidea and exploration of BIAs biosynthesis and accumulation based on UHPLC-MS/MS and combined sequencing platforms. Coptis deltoidea is an important medicinal plant with a long history of medicinal use, which is rich in benzylisoquinoline alkaloids (BIAs). In this study, Ultra performance liquid chromatography-electrospray ionization tandem mass spectrometry (UHPLC-ESI-MS/MS) and combined sequencing platforms were performed for exploration of BIAs biosynthesis, accumulation and comprehensive transcriptome analysis of C. deltoidea. By metabolism profiling, the accumulation of ten BIAs was analyzed using UHPLC-MS/MS and different contents were observed in different organs. From transcriptome sequencing result, we applied single-molecule real-time (SMRT) sequencing to C. deltoidea and generated a total of 75,438 full-length transcripts. We proposed the candidate biosynthetic pathway of tyrosine, precursor of BIAs, and identified 64 full length-transcripts encoding enzymes putatively involved in BIAs biosynthesis. RNA-Seq data indicated that the majority of genes exhibited relatively high expression level in roots. Transport of BIAs was also important for their accumulation. Here, 9 ABC transporters and 2 MATE transporters highly homologous to known alkaloid transporters related with BIAs transport in roots and rhizomes were identified. These findings based on the combined sequencing platforms provide valuable genetic information for C. deltoidea and the results of transcriptome combined with metabolome analysis can help us better understand BIAs biosynthesis and transport in this medicinal plant. The information will be critical for further characterization of C. deltoidea transcriptome and molecular-assisted breeding for this medicinal plant with scarce resources.

A Multi-Level Strategy Based on Metabolic and Molecular Genetic Approaches for the Characterization of Different Coptis Medicines Using HPLC-UV and RAD-seq Techniques.

Coptis plants (Ranunculaceae) to have played an important role in the prevention and treatment human diseases in Chinese history. In this study, a multi-level strategy based on metabolic and molecular genetic methods was performed for the characterization of four Coptis herbs (C. chinensis, C. deltoidea, C. omeiensis and C. teeta) using high performance liquid chromatography-ultraviolet (HPLC-UV) and restriction site-associated DNA sequencing (RAD-seq) techniques. Protoberberine alkaloids including berberine, palmatine, coptisine, epiberberine, columbamine, jatrorrhizine, magnoflorine and groenlandicine in rhizomes were identified and determined based on the HPLC-UV method. Among them, berberine was demonstrated as the most abundant compound in these plants. RAD-seq was applied to discover single nucleotide polymorphisms (SNPs) data. A total of 44,747,016 reads were generated and 2,443,407 SNPs were identified in regarding to four plants. Additionally, with respect to complicated metabolic and SNP data, multivariable statistical methods of principal component analysis (PCA) and hierarchical cluster analysis (HCA) were successively applied to interpret the structure characteristics. The metabolic variation and genetic relationship among different Coptis plants were successfully illustrated based on data visualization. Summarily, this comprehensive strategy has been proven as a reliable and effective approach to characterize Coptis plants, which can provide additional information for their quality assessment.

Comprehensive quality assessment for Rhizoma Coptidis based on quantitative and qualitative metabolic profiles using high performance liquid chromatography, Fourier transform near-infrared and Fourier transform mid-infrared combined with multivariate statistical analysis.

Rhizoma Coptidis (RC) originated from the dried rhizomes of Coptis herbal species is a widely used traditional Chinese medicine in history. In this study, a comprehensive quality assessment for RC medicines from C. chinensis, C. deltoidea, C. omeiensis and C. teeta species was performed based on quantitative and qualitative metabolic profiles obtained from high performance liquid chromatography (HPLC), Fourier transform near-infrared (FT-NIR) and Fourier transform mid-infrared (FT-MIR) combined with multivariate statistical analysis. Eight alkaloids including magnoflorine, groenlandicine, jatrorrhizine, columbamine, epiberberine, coptisine, palmatine and berberine were simultaneously identified and determined. Epiberberine, berberine, magnoflorine and groenlandicine were identified as possible index components. FT-NIR and FT-MIR profiles presented the holistic metabolic characterization of RC medicines. Principal component analysis (PCA) and hierarchical cluster analysis (HCA) were successively performed to clearly illustrate the metabolic variation and taxonomic relationship among four RC medicines. Additionally, taking berberine as an example, spectral quantification potential was investigated by referring HPLC data, using a conventional partial least squares regression (PLSR) algorithm. Data fusion strategy exhibited a better prediction for this compound than a single technique. Summary, these techniques can complement each other and provide a comprehensive and effective quality assessment for RC originated from different Coptis plants.