Ethnopharmacological relevance: Parkinson's disease (PD) is a rapidly progressing neurological disorder. Currently, Medication for PD has numerous limitations. Baichanting Compound (BCT) is a Chinese herbal prescription, a Combination of Acanthopanax senticosus (Rupr. and Maxim.) Harms, Paeonia lactiflora Pall and Uncaria rhynchophylla (Miq.) Miq. ex Havil, that was developed to treat PD and holds a national patent (ZL, 201110260536.3).Aim of the study: To clarify the therapeutic effect of BCT on PD and explore its possible mechanism based on metabolomics and metagenomics.Materials and methods: C57BL/6 mice were used as a control group, and alpha-syn transgenic C57BL/6 mice were randomly assigned to the PD (without treatment) or BCT (with BCT treatment) group. UPLC-MS was performed to detect dopamine levels in brain tissue, while ELISA was used to determine inflammatory factors such as IL-113, IL-6, TNF-alpha, IFN-gamma and NO, and oxidative stress indicators such as malondialdehyde, superoxide dismutase and glutathione peroxidase enzyme activity. Fecal metabolomics was used to detect fecal metabolic profiles, screen differential metabolic markers, and predict metabolic pathways by KEGG enrichment analysis. Metagenomics was used to determine the intestinal microbial composition, and KO enrichment analysis was performed to predict the potential function of different gut microbiota. Finally, Spearman correlation analysis was used to find the possible relationships among intestinal flora, metabolic markers, inflammatory factors, oxidative stress and dopamine levels.Results: BCT increased the superoxide dismutase activity of alpha-Syn transgenic C57BL/6 mice (P < 0.01), decreased the levels of TNF-alpha, IFN-gamma, IL-113, IL-6, NO and malondialdehyde (P < 0.01, 0.05), and increased the release of dopamine (P < 0.01). Metabolomics results show that BCT could regulate Acetatifactor, Marvinbryantia, Faeca-litalea, Anaeromassilibacillus, Anaerobium, Pseudobutyrivibrio and Lachnotalea and Acetatifactor_muris, Marvin-bryantia_formatexigens, Lachnotalea_sp_AF33_28, Faecalitalea_sp_Marseille_P3755 and Anaerobium_acetethylicum, Gemmiger_sp_An120 abundance to restore intestinal flora function, and reverse fecal metabolism trend, restoring the content of alpha-D-glucose, cytidine, L-glutamate, L-glutamine, N-acetyl-L-glutamate, raffinose and uracil. In addition, it regulates arginine biosynthesis, D-glutamine and D-glutamate, pyrimidine, galactose and alanine, aspartate and glutamate metabolic pathways.Conclusion: BCT may regulate the composition of the gut microbiota to reverse fecal metabolism in PD mice to protect the substantia nigra and striatum from oxidative stress and inflammatory factors and ultimately play an anti-PD role.

The incidence rate of Parkinson's disease (PD) is increasing yearly. Neuronal apoptosis caused by abnormal protein phosphorylation is closely related to the pathogenesis of Parkinson's disease. At present, few PD-specific apoptosis pathways have been revealed. To investigate the effect of Baichanting (BCT) on apoptosis from the perspective of protein phosphorylation, α-syn transgenic mice were selected to observe the behavioral changes of the mice, and the apoptosis of substantia nigra cells were detected by the HE method and TUNEL method. Network pharmacology combined with phosphorylation proteomics was used to find relevant targets for BCT treatment of PD and was further verified by PRM and western blotting. BCT improved the morphology of neurons in the substantia nigra and reduced neuronal apoptosis. The main enriched pathways in the network pharmacology results were apoptosis, the p53 signaling pathway and autophagy. Western blot results showed that BCT significantly regulated the protein expression levels of BAX, Caspase-3, LC3B, P53 and mTOR and upregulated autophagy to alleviate apoptosis. Using phosphorylated proteomics and PRM validation, we found that Pak5, Grin2b, Scn1a, BcaN, L1cam and Braf are closely correlated with the targets of the web-based pharmacological screen and may be involved in p53/mTOR-mediated autophagy and apoptosis pathways. BCT can inhibit the activation of the p53/mTOR signaling pathway, thereby enhancing the autophagy function of cells, and reducing the apoptosis of neurons which is the main mechanism of its neuroprotective effect.

High-polarity iridoids from Radix Scrophulariae (R. Scrophulariae) offer a range of benefits, including anti-inflammatory, antioxidant, antitumour, antibacterial, antiviral, and antiallergic effects. Although previous studies have indicated the potential of R. Scrophulariae for hyperthyroidism prevention and treatment, the specific active compounds involved and their mechanisms of action are not fully understood. This study explored the effects of high-polarity iridoid glycosides from R. Scrophulariae on hyperthyroidism induced in rats by levothyroxine sodium. The experimental design included a control group, a hyperthyroidism model group, and a group treated with iridoid glycosides. Serum triiodothyronine (T3) and thyroxine (T4) levels were quantified using an enzyme-linked immunosorbent assay (ELISA). Transcriptomic and proteomic analyses were applied to liver samples to identify differentially expressed genes and proteins. These analyses were complemented by trend analysis and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analysis. The effectiveness of key factors was further examined through molecular biology techniques. ELISA results indicated a notable increase in T3 and T4 in the hyperthyroid rats, which was significantly mitigated by treatment with iridoid glycosides. Transcriptomic analysis revealed 6 upregulated and 6 downregulated genes in the model group, showing marked improvement following treatment. Proteomic analysis revealed changes in 30 upregulated and 50 downregulated proteins, with improvements observed upon treatment. The PI3K-Akt signalling pathway was investigated through KEGG enrichment analysis. Molecular biology methods verified the upregulation of Spp1, Thbs1, PI3K, and Akt in the model group, which was reversed in the treatment group. This study revealed that highly polar iridoids from R. Scrophulariae can modulate the Spp1 gene and Thbs1 protein via the PI3K-Akt signalling pathway, suggesting a therapeutic benefit for hyperthyroidism and providing a basis for drug development targeting this condition.

Background/aims: Atherosclerosis (AS) is the common pathological basis of a variety of cardiovascular diseases (CVD), and has become the main cause of human death worldwide, and the incidence is increasing and younger trend. Ginsenoside Rb1 (Rb1), an important monomer component of the traditional Chinese herb ginseng, known for its ability to improve blood lipid disorders and anti-inflammatory. In addition, Rb1 was proved to be an effective treatment for AS. However, the effect of Rb1 on AS remains to be elucidated. The aim of this study was to investigate the mechanisms of Rb1 in ameliorating AS induced by high-fat diet (HFD).Materials and methodsIn this study, we developed an experimental AS model in Sprague-Dawley rats by feeding HFD with intraperitoneal injection of vitamin D3. The potential therapeutic mechanism of Rb1 in AS rats was investigated by detecting the expression of inflammatory factors, microbiome 16S rRNA gene sequencing, short-chain fatty acids (SCFAs) targeted metabolomics and untargeted metabolomics.ResultsRb1 could effectively alleviate the symptoms of AS and suppress the overexpression of inflammation-related factors. Meanwhile, Rb1 altered gut microbial composition and concentration of SCFAs characterized by Bacteroidetes, Actinobacteria, Lactobacillus, Prevotella, Oscillospira enrichment and Desulfovibrio depletion, accompanied by increased production of acetic acid and propionic acid. Moreover, untargeted metabolomics showed that Rb1 considerably improved faecal metabolite profiles, particularly arachidonic acid metabolism and primary bile acid biosynthesis.ConclusionRb1 ameliorated the HFD-induced AS, and the mechanism is related to improving intestinal metabolic homeostasis and inhibiting systemic inflammation by regulating gut microbiota.

ObjectiveThis study integrates network pharmacology and metabolomics techniques to explore the potential regulatory mechanisms of Res on gouty arthritis (GA).MethodsNetwork pharmacology was used to predict the mechanism of Res in regulating GA, and methods such as HE staining, ELISA, immunohistochemistry, Real-time PCR, Western blot, and molecular docking were used to verify the role of NF-κB, MAPK, and JAK/STAT inflammatory signaling pathways in the MSU-induced GA rat model. In addition, non-targeted metabolomics techniques were combined to further investigate the mechanism of Res in treating GA.ResultsThe results of network pharmacology showed that Res may exert its therapeutic effects through the NF-κB signaling pathway. Animal experiments demonstrated that in the MSU-induced GA rat model, pathological damage, serum biochemical indicators, and levels of inflammatory factors were significantly increased, and the NF-κB signaling pathway was activated. The intervention of Res significantly reduced pathological damage, serum biochemical indicators, levels of inflammatory factors, and the activation of NF-κB, MAPK, and JAK/STAT signaling pathways in the model rats. Metabolomics results showed that Res could improve the metabolic trajectory deviations in serum and joint fluid of GA model rats. Through related metabolic pathway analysis, the most affected metabolic pathways were found to be Sphingolipid metabolism, Glycerophospholipid metabolism, Phenylalanine, tyrosine and tryptophan biosynthesis, Pantothenate and CoA, Citrate cycle (TCA cycle), and Arachidonic acid metabolism.ConclusionResveratrol can regulate the biosynthetic pathways of arachidonic acid, phenylalanine, tyrosine, and tryptophan, pantothenic acid and CoA biosynthesis pathways, TCA cycle, and other metabolic pathways, thereby regulating the NF-κB, MAPK, and JAK/STAT3 signaling pathways, and inhibiting the acute inflammatory response during GA attacks, showing characteristics of multi-pathway and multi-target action.

Parkinson's disease (PD) is the second most common neurodegenerative disease, with an increasing prevalence as the population ages, posing a serious threat to human health, but the pathogenesis remains uncertain. Acanthopanax senticosus (Rupr. et Maxim.) Harms (ASH) (aqueous ethanol extract), a Chinese herbal medicine, provides obvious and noticeable therapeutic effects on PD. To further investigate the ASH's mechanism of action in treating PD, the structural and functional gut microbiota, as well as intestinal metabolite before and after ASH intervention in the PD mice model, were examined utilizing metagenomics and fecal metabolomics analysis. α-syn transgenic mice were randomly divided into a model and ASH groups, with C57BL/6 mice as a control. The ASH group was gavaged with ASH (45.5 mg/kg/d for 20d). The time of pole climbing and autonomous activity were used to assess motor ability. The gut microbiota's structure, composition, and function were evaluated using Illumina sequencing. Fecal metabolites were identified using UHPLC-MS/MS to construct intestinal metabolites. The findings of this experiment demonstrate that ASH may reduce the climbing time of PD model mice while increasing the number of autonomous movements. The results of metagenomics analysis revealed that ASH could up-regulated Firmicutes and down-regulated Actinobacteria at the phylum level, while Clostridium was up-regulated and Akkermansia was down-regulated at the genus level; it could also recall 49 species from the phylum Firmicutes, Actinobacteria, and Tenericutes. Simultaneously, metabolomics analysis revealed that alpha-Linolenic acid metabolism might be a key metabolic pathway for ASH to impact in PD. Furthermore, metagenomics function analysis and metabolic pathway enrichment analysis revealed that ASH might influence unsaturated fatty acid synthesis and purine metabolism pathways. These metabolic pathways are connected to ALA, Palmitic acid, Adenine, and 16 species of Firmicutes, Actinobacteria, and Tenericutes. Finally, these results indicate that ASH may alleviate the movement disorder of the PD model, which may be connected to the regulation of gut microbiota structure and function as well as the modulation of metabolic disorders by ASH.

Gouty arthritis (GA) is an inflammatory disorder that is associated with elevated serum levels of uric acid. Total saponins from Dioscorea nipponica Makino (TSDN) are a natural component that ameliorates inflammation while also decreasing uric acid levels. The aim of the present study was to unravel the mechanism of TSDN in gouty rats in regard to regulation of the formation of neutrophil extracellular traps (NETs) via the PI3K/AKT/mTOR axis. A total of 40 Wistar rats were divided into 4 groups: normal, model, TSDN and rapamycin groups. Reverse-transcription-quantitative PCR (RT-qPCR) and western blot analysis were used to assess the mRNA and protein expression levels of the PI3K/AKT/mTOR axis. The formation of NETs was detected by immunohistochemical and immunofluorescent methods. ELISA was used to measure the levels of IL-1β and TNF-α. RT-qPCR and western blotting demonstrated that TSDN compromised the mRNA and protein expression levels of activated protein kinase (AMPK) and mTOR, as well as the mRNA expression levels of AKT and PTEN. Furthermore, it increased the protein expression levels of phosphorylated (p-) PI3K, p-AKT and p-AMPK. Immunohistochemical and immunofluorescent analyses revealed that TSDN decreased the protein expression levels of neutrophil elastase, proteinase 3, cathepsin G, lactoferrin and myeloperoxidase, as well as the number of citrullinated histone 3+ cells. TSDN also reduced the release of IL-1β and TNF-α. Overall, the anti-inflammatory action of TSDN in gouty rats may be realized by suppressing the formation of NETs by regulating the PI3K/AKT/mTOR axis.

Abstract Objective:This study investigated the regulatory mechanism of the NLRP3/GSDMD pathway in GA pyroptosis using network pharmacology and in vitro and in vivo experiments. Methods:In this study, network pharmacology was used to predict the mechanism of pyroptosis in Gouty arthritis, and HE, ELISA, Real-time PCR, Western blot, immunoprecipitation, and flow cytometry were used to further investigate the mechanism of NLRP3/GSDMD signal axis in GA rat model induced by MSU and THP-1 cells induced by MSU. Results:When gout arthritis pyroptosis occurred, the NOD-like receptor pathway was activated, as determined by network pharmacology. In the rat model of GA induced by MSU, the swelling degree, pathological damage, serum biochemical indices, and inflammatory factors of the joints were significantly increased, and the expressions of key pyroptosis proteins such as NLRP3, GSDMD, ASC, Caspase-1, and IL-1β were significantly increased in ankle joints, indicating the existence of a NOD-like receptor pathway mediating pyroptosis during the acute attack of GA. The results of NLRP3 and GSDMD inhibitor intervention showed that inhibitor intervention significantly inhibited the expression of pyroptosis protein. At the same time, it was found that the activation of NLRP3 inflammatory bodies was inhibited, and the expressions of GSDMD, ASC, Caspase-1 and IL-1β protein were down-regulated, but the expression of NLRP3 protein was not affected by inhibiting the activation of GSDMD and down-regulating the expression of IL-1β. The results of the cell experiment revealed that the inhibitor group prevented MSU-induced Nod-like receptor-mediated pyroptosis in THP-1. Co-precipitation results also revealed a close interaction between NLRP3 and GSDMD. Conclusion:Inhibiting the NLRP3/GSDMD signaling axis can further reduce the occurrence of pyroptosis and thus alleviate the inflammatory response in acute Gouty arthritis attacks. Through a Caspase-1-dependent mechanism, NLRP3 regulates the process of pyroptosis in the occurrence of Gouty arthritis.

Gouty arthritis (GA) cause great harm to patients. Cellular pyroptosis, a mode of programmed cell death associated with inflammatory response, is closely related to GA. Both cysteamine aspartate-1-dependent and non-dependent pathways are involved in the progression of GA. During GA development, high blood uric acid levels leads to excessive biologically-inspired NOD-like receptor thermal protein domain associated protein 3 (NLRP3) inflammasome activation to drive caspase-1 activation for promoting the maturation of interleukin-1β precursors, and caspase-1 activation disrupts the amino terminus in gasdermin D-N (GSDMD-N) and carboxy-terminal gasdermin-C structural domains, causing pores in the membrane and thus inducing the onset of scorch death. Therefore, modulating the onset of scorch death may become an important target for drug intervention in diseases. Chinese medicine is substantially biologically inspired and used synergistically to treat GA through multiple pathways and targets, which may regulate the relevant pathways through cellular pyroptosis quality. This study focuses on the interpretable regulatory mechanism of cellular pyroptosis bionic in GA and the role of Chinese medicine on GA, which provides a new scientific basis and strategy for targeting cellular pyroptosis bionic as the prevention and treatment quality of GA.

Huangqi Chifeng Decoction (HQCFT), a traditional Chinese medicine preparation, has long been used to treat cardiovascular and cerebrovascular diseases. However, the mechanism of the beneficial effect of HQCFT on atherosclerosis remains to be explored. In this work, to investigate the effects of HQCFT on bile acid (BA) metabolism and the gut microbiome in atherosclerosis, ApoE−/− mice were fed a with high-fat diet for 16 weeks to establish the AS model. HQCFT(1.95 g kg−1 and 3.9 g kg−1 per day) was administered intragastrically for 8 weeks to investigate the regulatory effects of HQCFT on gut microbiota and bile acid metabolism and to inhibit the occurrence and development of AS induced by a high-fat diet. Histopathology, liver function and blood lipids were used to assess whether HQCFT can reduce plaque area, regulate lipid levels and alleviate liver steatosis in AS mice. In addition, 16S rDNA sequencing was used to screen the gut microbiota structure, and ultrahigh-performance liquid chromatography-tandem mass spectrometry (UPLC‒MS/MS) was used to determine the bile acid profile. The mRNA and protein expression levels of bile acid metabolism were detected by RT‒PCR and WB to find the potential correlation. Results: HQCFT can regulate gut microbiota disorders, which was achieved by increasing gut microbiota diversity and altering Proteobacteria, Desulfobacterota, Deferribacteres, Rodentibacter, Parasutterella, and Mucispirillum interference abundance to improve AS-induced gut microbiota. HQCFT can also adjust the content of bile acids (TCA, LCA, DCA, TDCA, TLCA, UDCA, etc.), regulate bile acid metabolism, relieve liver fat accumulation, and inhibit the process of AS. In addition, HQCFT can restore the abnormal metabolism of bile acid caused by AS by regulating the expression of farnesoid X receptor (FXR), liver X receptor α (LXRα), ABCA1, ABCG1 and CYP7A1. Conclusion: HQCFT may play a part in the prevention of atherosclerosis by inhibiting the FXR/LXRα axis, increasing the expression of CYP7A1 in the liver, and regulating the interaction between the gut microbiota and bile acid metabolism.

Ethnopharmacological relevance: Integrated Chinese herbal medicine (CHM) and Western Medicine (WM) treat-ments have been used for primary hypertension (PHTN) patients in China. Currently, there are many randomized control trials (RCTs) published regarding the effect of CHM and WM on PHTN, which indicated that combining Chinese with WM was effective and safe for PHTN when compared with WM alone, but the quality of evidence was insufficient, and there is no clear information and summary are available for these RCTs assessing the effectiveness of CHM with WM versus WM in patients with PHTN.Objectives: This systematic study and meta-analysis aimed to evaluate the effectiveness and safety of CHM combined with WM in comparison with WM in reducing systolic and diastolic blood pressure for patients with PHTN. Methods: The information of this study was searched from electronic databases (PubMed, COCHRANE, EMBASE, Ovid, CNKI, VIP, Wanfang, and CBM). The markedly effective and effective terms were according to Guiding Principles for Clinical Research of New Chinese Medicines. Two investigators independently reviewed each trial. The Cochrane risk of bias assessment tool was used for quality assessment, and RevMan 5.4 was used for meta -analysis.Results: In this study, a total of 29 studies that included 2623 patients were recorded. The study results displayed that the clinical effectiveness in the treatment of hypertension patients from the integrated medicines was considerably higher than that with WM alone, clinical effective (RR 1.23, 95% CI [1.17, 1.30], P < 0.00001), and markedly effective (ME) in the patients (RR 1.66, 95% CI [1.52, 1.80], and P < 0.00001). Random effect in SBP (MD 7.91 mmHg,[6.00, 983], P < 0.00001) and DBP (MD 5.46 mmHg, [3.88, 6.43], P < 0.00001), a subgroup analysis was carried out based on the type of intervention, duration of treatment, and CHM formulas that showed significance. Furthermore, no severe side effects were reported, and no patients stopped treatment or withdrawal due to any severe adverse events. Conclusion: Compared to WM alone, the therapeutic effectiveness of CHM combined with WM is significantly improved in the treatment of hypertension. Additionally, CHM with WM may safely and efficiently lower systolic blood pressure (SBP) and diastolic blood pressure (DBP) in individuals with PHTN. However, rigorous ran-domized controlled trials with a large sample, high quality, long duration of treatment, and follow-up are rec-ommended to strengthen this clinical evidence.

Background: The deposition of MSU crystals within joints and surrounding soft tissues can potentially elicit the onset of GA. While there have been investigations regarding the inflammatory role of microRNA in gout disease, research on the impact of PMN-Exo on the inflammation of the GA joint is scarce.Objective: The present study aims to isolate and extract PMN-Exo for subsequent in situ infusion testing. The objective was to investigate the impact of PMN-Exo on GA joint injury.Method: The microRNA sequencing was performed on the joint tissue and PMN-Exo to screen DE-miRNAs and predict their respective target gene and function.Results: TEM, WB, and NTA results show that the extracted PMN Exo conforms to the characteristics of exosomes. The reinfusion experiment revealed that PMN-Exo could facilitate an increase in inflammatory factor concentration (IL-1β, IL-8, and TNF-α) and ultimately contribute to GA joint injury. The ankle tissue sequencing in this experiment revealed the co-expression of 18 DE-miRNAs. They were primarily concentrated on regulating cell proliferation or migration and regulating signaling pathways such as mTOR, Ras, and Wnt. Similarly, the PMN-Exo indicated the presence of 254 DE-mRNAs in control vs model-PMN-Exo group. They mainly regulated actin cytoskeletal tissue, Rap1, oxytocin, and BCR signaling pathways. These findings offer insights into potential regulatory mechanisms involved in PMN-Exo-mediated GA joint injury. After integrating and analyzing, six core miRNAs were identified, which play a crucial role in regulating actin cytoskeleton, Rap1, and Ras signaling pathways. These core genes included rno-miR-204-3p, rno-miR-1896, rno-miR-30c-1-3p, rno-miR-92a-2-5p, rno-miR-6328, and rno-miR-667-3p.Conclusion: It can be asserted that PMN-Exo can enhance the damage to joints caused by Gouty arthritis and augment the associated inflammatory response. Based on the results of high-throughput sequencing and data integration analysis, 6 core miRNAs may play a role in the process of PMN-Exo promoting joint injury by regulating the activity of actin cytoskeleton and the Rap1 and Ras signal pathways.

Objective. Using network pharmacology and gut microbiota sequencing to investigate the probable mechanism of Huangqin decoction in the treatment of Diabetic enteritis (DE). Methods. The mechanism of Huangqin decoction on DE was studied by combining network pharmacology and gut microbiota sequencing analysis. The core components and possible targets of Huangqin decoction were analyzed by network pharmacology. The effect of Huangqin decoction on microorganisms was investigated by gut microbiota sequencing. Results. The results of gut microbiota sequencing analysis showed the abundance of TM7, Tenericutes, Chloroflexi, Cyanobacteria, Acidobacteria, WS6, [Prevotella], Helicobacter, Prevotella, Lactococcus, and Anaeroplasma in the Huangqin decoction group had a significant downward. Using a network pharmacology-related database, 141 main active components of Huangqin decoction were identified, as well as 256 corresponding component targets and 1777 corresponding disease targets; the disease targets and component targets were mapped, and topological analysis was used to determine the potential of Huangqin decoction in the treatment of DE. There were 156 targets, of which the top 20 genes were selected for GO and KEGG. The KEGG results showed that 134 pathways were enriched, which was partially consistent with the metabolic pathways of gut microbiota sequencing analysis. Conclusion. The results show that Huangqin decoction can inhibit the expression of inflammatory factors and related inflammatory pathways in intestinal epithelial cells, thereby regulating the structure of intestinal flora. Using picurst2 for functional prediction and metabolic pathway statistics, seven metabolic pathways were obtained consistent with gut microbiota sequencing, and the NOD-like receptor signaling pathway may be its potential molecular mechanism. These results help to understand the mechanism of Huangqin decoction on DE and provide the theoretical basis for further study of Huangqin decoction.

Primary hypertension is understood as a disease with diverse etiology, a complicated pathological mechanism, and progressive changes. Gedan Jiangya Decoction (GJD), with the patent publication number CN114246896A, was designed to treat primary hypertension. It contains six botanical drugs; however, the underlying mechanism is uncertain. We utilized network pharmacology to predict the active components, targets, and signaling pathways of GJD in the treatment of primary hypertension. We also investigated the potential molecular mechanism using molecular docking and animal experiments. The Traditional Chinese Medicine System Pharmacology Database and Analysis Platform (TCMSP), the Protein Database (UniProt), and a literature review were used to identify the active components and related targets of GJD's pharmacological effects. The GeneCards, Online Mendelian Inheritance in Man (OMIM), Therapeutic Target Database (TTD), and DrugBank databases were utilized to identify hypertension-related targets. Based on a Venn diagram of designed intersection targets, 214 intersection targets were obtained and 35 key targets for the treatment of hypertension were determined using the STRING data platform and Cytoscape software. The Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analysis of key targets revealed that the relevant molecular action pathways of GJD in the treatment of hypertension include the Toll-like receptor, MAPK, PI3K-Akt, and renin-angiotensin signaling pathways. A GJD active ingredient-key target-pathway connection diagram was created using Cytoscape software, and 11 essential active components were selected. Molecular docking was then used to verify the binding activity of key targets and key active ingredients in GJD to treat primary hypertension. The results of this study indicate that AGTR1, AKT1 with puerarin, EDNRA with tanshinone IIA, MAPK14 with daidzein, MAPK8 with ursolic acid, and CHRM2 with cryptotanshinone had high binding activity to the targets with active components, whereas AGTR1 was selected as target genes verified by our experiment. HPLC was utilized to identify the five active ingredients. Experiments in high-salt rats demonstrated that GJD might decrease the expression of AGTR1 in the kidney and thoracic aorta while increasing the expression of eNOS by preventing the activation of the renin-angiotensin pathway, thereby reducing lowering systolic and diastolic blood pressure.

Gedan Jiangya decoction (GJD) (aqueous ethanol extract), a traditional Chinese medicine formula which contain six botanical drugs (Uncaria rhynchophylla (Miq.) Miq., Salvia miltiorrhiza Bunge, Pueraria lobata (Willd.) Ohwi, Eucommia ulmoides Oliv., Prunella vulgaris L., and Achyranthes bidentata Blume) was designed to treat hypertension; however, the underlying mechanism of action is unclear. This study aimed to determine the mechanisms of action of GJD in the treatment of hypertension in spontaneously hypertensive rats (SHR). Male SHRs were randomly divided into five groups: GJD doses were low (1.36 g/kg/d), medium (2.72 g/kg/d), and high (5.44 g/kg/d), captopril (13.5 mg/kg/d), and SHR groups, with Wistar-Kyoto rats (WKY) serving as the control. Every rat was gavaged once a day. The ALC-NIBP, a noninvasive blood pressure device, measured systolic (SBP) and diastolic (DBP) blood pressures. Six weeks following treatment, all rats were anesthetized. The blood samples were obtained from the abdominal aorta and then serum isolated to assess endothelin-1 and angiotensin II, interleukin-1beta, interleukin-6, and TNF-alpha. The left ventricular and thoracic aortas were taken for HE staining, immunohistochemistry, RT-qPCR, and western blot examination. Following GJD therapy, SBP and DBP were significantly lowered, as were serum levels of endothelin-1 and angiotensin II. The thickness of the left ventricular and thoracic aorta walls reduced, as did type I collagen, type III collagen, and alpha-SMA expression in the left ventricular and aortic tissues. The GJD treatment significantly reduced serum levels of the inflammatory markers interleukin-1beta, interleukin-6, and TNF-alpha. Furthermore, interleukin-1 beta, interleukin-6, TNF-alpha, TAK1, and NF-κB/p65 levels were significantly reduced in left ventricular and aortic tissues, whereas IkB-alpha levels were significantly elevated. GJD has a dose-dependent effect on all parameters. In conclusion, GJD has been shown to lower blood pressure, improve cardiovascular remodeling, and reduce inflammation via regulating NF-κB in SHRs.

Objective. The aims of this study were to investigate the impact of TAK-242 on the Toll-like receptor 4 (TLR4)/myeloid differentiation factor 88 (MyD88)/nuclear transcription factor-κB (NF-κB) signal transduction pathway in rats with hepatic fibrosis (HF) using the liver gut axis and to investigate the molecular mechanism of its intervention on HF. Methods. SPF grade SD male rats were randomly allocated to the control, model, and TAK-242 groups. For 8 weeks, the model and TAK-242 groups received 3 mL·kg-1 (the initial dose 5 mL·kg-1) intraperitoneal injections of 40% CCL4 olive oil solution. TAK-242 (5 mg·kg-1) was administered once a day for 5 days after modeling. The pathological alterations of liver and small intestine tissues in each group were observed using H&E and Masson staining. ELISA was used to measure serum levels of alanine aminotransferase (ALT), aspartate aminotransferase (AST), direct bilirubin (DBIL), total bilirubin (TBIL), interleukin-1β (IL-1β), interleukin-6 (IL-6), and tumor necrosis factor alpha (TNF-α). RT-qPCR was utilized to identify the mRNA expression level of IL-1β, IL-6, TNF-α, TLR4, MyD88, and NF-κB in rat liver and small intestine tissues. The protein level of IL-1β, IL-6, TNF-α, TLR4, MyD88, and NF-κB protein in rat liver and small intestine tissues was determined utilizing Western blot and IHC. Results. TAK-242 significantly reduced AST, ALT, TBIL, and DBIL expression in HF rats’ serum ( P < 0.01 ) and alleviated liver tissue injury. Hematoxylin-eosin (H&E) and Masson staining revealed inflammatory cell infiltration and fibrous proliferation in the liver and small intestine tissue in the model group and partial cell swelling in the TAK-242 group, which indicated a considerable improvement compared to the model group. RT-qPCR, Western blot, and IHC data indicated that TAK-242 reduced the IL-1β, IL-6, TNF-α, TLR4, MyD88, and NF-κB expression in the liver and small intestine tissues of HF rats. Conclusion. TAK-242 might downregulate the TLR4/MyD88/NF-κB signal pathway through the liver-gut axis, suppress the inflammatory response, and eventually alleviate HF in rats.

Neutrophil extracellular traps(NETs) are networks of extracellular fibers primarily composed of DNA, histones, granular proteins, and cytoplasmic proteins and released to the outside of cells by neutrophils under the stimulation of bacteria, fungi, viruses, parasites, etc. NETs are generated in two forms, suicidal NETs and vital NETs, according to different stimuli. NETs have both anti-inflammatory and pro-inflammatory effects. On the one hand, they can play the anti-microbial role to resist inflammation by capturing, fixing, and killing invading pathogens, which is a special way for neutrophils to exert host defenses. On the other hand, in case of excessive formation or insufficient elimination, they can cause tissue damage directly, and also promote the release of inflammatory factors by recruiting other pro-inflammatory cells or proteins to further expand the inflammatory response, which is related to the pathologies of many diseases. In autoimmune diseases, NETs as important sources of autoantigens, can act as danger-associated molecular patterns( DAMPs) and activate the nucleotide-binding oligomerization domain leucine-rich repeats containing pyrin domain 3(NLRP3) inflammasome and complement system, thereby breaking self-tolerance and accelerating autoimmune inflammation. In addition, NETs can also activate other immune cells(such as B cells, antigen-presenting cells, and T cells) and regulate the acquired immune response. The present study reviewed the correlation of NETs with diseases such as systemic lupus erythematosus(SLE), rheumatoid arthritis(RA), and gouty arthritis(GA) to reveal the effect of dynamic balance between formation and clearance of NETs in autoimmune diseases and provide a theoretical basis for the investigation of underlying mechanisms and targeted therapies of traditional Chinese medicine.

Radix Scrophulariae (Chinese name: Xuanshen), a traditional Chinese herb, is used for the treatment of hyperthyroidism, and in this study, its mechanisms were evaluated by metabonomics and system pharmacology. To study the anti-hyperthyroidism effects of R. Scrophulariae, a male SD rat (180–220 g) model of hyperthyroidism induced by Euthyrox was used. Thirty rats were randomly distributed into three groups: the Model group, the R. Scrophulariae treatment group (RS group) and the healthy Control group. Using the UHPLC/Q-TOF-MS metabolomics approach, 44 metabolites were found to be profoundly altered in the model group, and the levels of these biomarkers were significantly decreased after treatment with R. Scrophulariae. Forty-four metabolites and 13 signaling pathways related to R. Scrophulariae, including the biosynthesis of unsaturated fatty acids, primary bile acid biosynthesis and sphingolipid metabolism, were explored, and linoleic acid metabolism and sphingolipid metabolism were identified as the most relevant metabolic pathways. In addition, the system pharmacology paradigm revealed that R. Scrophulariae contains 83 active ingredients and is related to 795 genes, and 804 disease genes are related to hyperthyroidism. The construction of the R. Scrophulariaceae-chemical composition-target-hyperthyroidism network identified a total of 112 intersection genes. The enriched gene targets were analyzed, and five pathways were found to be enriched. Among them pathways, the HIF signaling pathway had the highest enrichment score, which indicated that this pathway might be the main signaling pathway related to the treatment of hyperthyroidism by R. Scrophulariae.The integrated approach involving metabolomics and network pharmacology revealed that R. Scrophulariae might play a role in the treatment of hyperthyroidism by regulating the “IL6-APOA1-cholesterol” pathway and disturbing the HIF signaling pathway. The results demonstrate that the combination of metabolomics and network pharmacology could be used to reflect the effects of R. Scrophulariae on the biological network and metabolic state of hyperthyroidism and to evaluate the drug efficacy of R. Scrophulariaceae and its related mechanisms.

Diosbulbin-B (DB) is a promising therapeutic drug for cancer treatment; however, DB-induced hepatotoxicity seriously limits its clinical utilization. Based on this, the present study investigated whether the Angelica sinensis extract, angelica sinensis polysaccharide (ASP), was effective to attenuate DB-induced cytotoxicity in hepatocytes. The primary hepatocytes were isolated from rats and cultured in vitro, which were subsequently treated with high-dose DB (100 μM) and ASP (12 μg/ml) to establish the DB-induced hepatotoxicity models. MTT assay and flow cytometry (FCM) were performed to evaluate cell viability, and the results showed that high-dose DB-induced cell apoptosis and inhibition of proliferation were reversed by co-treating cells with ASP, which were supported by our Western Blot assay data that ASP upregulated Cyclin D1 and CDK2 to abrogate high-dose DB-induced cell cycle arrest. In addition, ASP exerted its regulating effects on cell autophagy, and we found that ASP increased LC3B-II/I ratio and Atg5, but decreased p62 to activate the autophagy flux. Of note, the MEK/ERK pathway could be activated by ASP in the DB-treated hepatocytes, and the protective effects of ASP on high-dose DB-induced hepatocyte death were abolished by co-treating cells with the autophagy inhibitor (3-methyladenine, 3-MA) and MEK/ERK selective inhibitor (SCH772984). Moreover, blockage of the MEK/ERK pathway suppressed cell autophagy in the hepatocytes co-treated with ASP and high-dose DB. Taken together, this in vitro study illustrated that ASP activated the MEK/ERK pathway mediated autophagy to suppress high-dose DB-induced hepatotoxicity.

<span id="ChDivSummary" name="ChDivSummary" class="abstract-text">巨噬细胞是人体重要的固有免疫细胞之一,有高度异质性和可塑性,在不同微环境刺激下可极化为M1型和M2型。巨噬细胞M1/M2表型与多种疾病的病理过程相关,但具体机制尚不明确。本文旨在对与其相关的疾病,如肿瘤、代谢性疾病、炎症性疾病进行描述,突出M1/M2极化稳态平衡在疾病中的重要作用,为临床治疗提供新靶点及方向。</span>