Berberine is a plant alkaloid that can be extracted from many Chinese herbs. It has been reported that berberine could protect mice from ulcerative colitis, but the mechanism remains unclear. The current study’s aim was to determine the potential mechanism by which berberine exhibits its anti-inflammatory function. Mice with colitis induced by dextran sulfate sodium (DSS) were administered with berberine at 50 mg/kg by gavage. Berberine significantly increased the proportion of regulatory T cells (Treg cells). The targeted metabolomics analysis was then performed to find that glutamine and glutamate metabolism played an important role in the process of regulating immune response. mTORC1 pathway was reported to closely relate with glutamine metabolism. As a result, the relative expression levels of downstream effector genes of mTORC were further determined, and data obtained showed that berberine could significantly increase the relative expression levels of S6K1 and 4EBP1. In addition, rapamycin was used to inhibit mTORC1 signaling, and it was found that colon length, disease associated index (DAI), and proportion of Treg cells of mice in the rapamycin-DSS group were not different from those of mice in the rapamycin/berberine-DSS group. Together, these results suggest that berberine exhibits significant protective effects against DSS colitis by activating the mTORC1 pathway to increase the proportion of Treg cells.

Introduction. As a novel global epidemic, corona virus disease 2019 (COVID-19) caused by SARS-CoV-2 brought great suffering and disaster to mankind. Recently, although significant progress has been made in vaccines against SARS-CoV-2, there are still no drugs for treating COVID-19. It is well known that traditional Chinese medicine (TCM) has achieved excellent efficacy in the treatment of COVID-19 in China. As a treasure-house of natural drugs, Chinese herbs offer a promising prospect for discovering anti-COVID-19 drugs. Hypothesis/Gap Statement. We proposed that Rhei Radix et Rhizome−Schisandrae Sphenantherae Fructus (RS) may have potential value in the treatment of COVID-19 patients by regulating immune response, protecting the cardiovascular system, inhibiting the production of inflammatory factors, and blocking virus invasion and replication processes. Aim. We aimed to explore the feasibility and molecular mechanisms of RS against COVID-19, to provide a reference for basic research and clinical applications. Methodology. Through literature mining, it is found that a Chinese herbal pair, RS, has potential anti-COVID-19 activity. In this study, we analysed the feasibility of RS against COVID-19 by high-throughput molecular docking and molecular dynamics simulations. Furthermore, we predicted the molecular mechanisms of RS against COVID-19 based on network pharmacology. Results. We proved the feasibility of RS anti-COVID-19 by literature mining, virtual docking and molecular dynamics simulations, and found that angiotensin converting enzyme 2 (ACE2) and 3C-like protease (3 CL pro) were also two critical targets for RS against COVID-19. In addition, we predicted the molecular mechanisms of RS in the treatment of COVID-19, and identified 29 main ingredients, 21 potential targets and 16 signalling pathways. Rhein, eupatin, (-)-catechin, aloe-emodin may be important active ingredients in RS. ALB, ESR1, EGFR, HMOX1, CTSL, and RHOA may be important targets against COVID-19. Platelet activation, renin secretion, ras signalling pathway, chemokine signalling pathway, and human cytomegalovirus infection may be important signalling pathways against COVID-19. Conclusion. RS plays a key role in the treatment of COVID-19, which may be closely related to immune regulation, cardiovascular protection, anti-inflammation, virus invasion and replication processes.

Artesunate is a widely used drug in clinical treatment of malaria. The aim of this study was to investigate the therapeutic mechanism of artesunate on malaria using an integrated strategy of network pharmacology and serum metabolomics. The mice models of malaria were established using 2 × 10⁷ red blood cells infected with Plasmodium berghei ANKA injection. Giemsa and hematoxylin-eosin (HE) staining were used to evaluate the efficacy of artesunate on malaria. Next, network pharmacology analysis was applied to identify target genes. Then, a metabolomics strategy has been developed to find the possible significant serum metabolites and metabolic pathways induced by artesunate. Additionally, two parts of the results were integrated to confirm each other. Giemsa and HE staining results showed that artesunate significantly inhibited the proliferation of Plasmodium and reduced liver and spleen inflammation. Based on metabolomics, 18 differential endogenous metabolites were identified as potential biomarkers related to the artesunate for treating malaria. These metabolites were mainly involved in the relevant pathways of biosynthesis of unsaturated fatty acids; aminoacyl-tRNA biosynthesis; valine, leucine, and isoleucine biosynthesis; and phenylalanine, tyrosine, and tryptophan biosynthesis. The results of the network pharmacology analysis showed 125 potential target genes related to the treatment of malaria with artesunate. The functional enrichment was mainly associated with lipid and atherosclerosis; pathways of prostate cancer and proteoglycans in cancer; and PI3K-Akt, apoptosis, NF-κB, Th17 cell, and AGE-RAGE signaling pathways. These findings were partly consistent with the findings of the metabolism. Our results further suggested that artesunate could correct the inflammatory response caused by malaria through Th17 cell and NF-κB pathways. Meanwhile, our work revealed that cholesterol needed by Plasmodium berghei came directly from serum. Cholesterol and palmitic acid may be essential in the growth and reproduction of Plasmodium berghei. In summary, artesunate may have an effect on anti-malarial properties through multiple targets.

Mulberroside A is a polyhydroxylated stilbene active component of Morus alba L. Studies have shown that it has antitussive, antiasthmatic, tyrosinase and antioxidation activities. However, little is known about the metabolism of it in vitro and in vivo. In our study, an integrated strategy on the basis of UHPLC-Q-Exactive Plus Orbitrap MS and network pharmacology was established to comprehensively research the metabolic characteristic of mulberroside A for the first time. Plasma, urine, feces and liver tissues of rats in the blank group and drug group were collected after intragastric administration of mulberroside A at a dose of 150 mg/kg, and rat liver microsomes were cultured for in vitro metabolism experiment. The biological samples were processed by different methods and analyzed in positive and negative ion modes using UHPLC-Q-Exactive Plus Orbitrap MS. A total of 72 metabolites were finally identified based on the accurate molecular mass, retention time, MS/MS spectra and related literatures combined with the Compound Discoverer 3.1. The metabolic pathways were mainly hydrolysis, glucuronidation, hydrogenation, sulfation, hydroxylation, methylation and their composite reactions. In addition, a network pharmacology method was used to predict the mechanism of action of mulberroside A and its metabolites. In the end, 7 metabolites with high gastrointestinal absorption and drug-likeness and 167 targets were screened by Swiss ADME and Swiss Target Prediction. 1702 items of GO analysis and 158 related signaling pathways of KEGG were enriched using Metascape. This study established a novel integrated strategy based on UHPLC-Q-Exactive Plus Orbitrap MS and network pharmacology, which could systematically analyze the metabolism behavior of mulberroside A in vivo and in vitro of rats and provide basis for the further research of mulberroside A.

Yinqiao powder, with significant anti‐inflammatory and antiviral effects, is a classical formula for the treatment of febrile diseases in China. During the SARS period in 2003, Yinqiao powder showed a good antipyretic effect. It also plays a major role in the treatment for COVID‐19 in China. Although there are many studies on the chemical compositions and pharmacological effects of Yinqiao powder, there are few studies on the quality standard system of it. In our study, a systematic quality evaluation method of Yinqiao powder combining HPLC fingerprint with quantitative analysis of multi‐components by single marker (QAMS) based on network pharmacology and UPLC‐Q‐Exactive‐Orbitrap‐MS was established for the first time. In the UPLC‐Q‐Exactive‐Orbitrap‐MS experiment, a total of 53 compounds were identified in the extract solution of Yinqiao powder. In addition, 33 blood components were characterized, 23 of which were prototypes. The results of network pharmacology analysis showed that Yinqiao powder may inhibit inflammatory responses by suppressing IL‐6, CXCL2, TNFα, NF‐κB, etc., in the treatment of COVID‐19. The HPLC fingerprint analysis of Yinqiao powder was conducted at 237 nm and 29 characteristic peaks were matched, 11 of which were identified. Forsythoside A was selected as the internal standard reference and double‐wavelength (237 nm and 327 nm) was established in QAMS experiment. The repeatability was well under different conditions, and the results measured by QAMS were consisted with that of the external standard method (ESM), indicating that the QAMS method was reliable and accurate. The quality evaluation method of Yinqiao powder would be helpful to evaluate the intrinsic quality of Yinqiao powder more comprehensively, which is conducive to improve the quality standard of Yinqiao powder and provide a beneficial guarantee for the clinical treatment of COVID‐19.