Chinese Traditional and Herbal Drugs

Shouhui Tongbian Capsule alleviate post-stroke depression by modulating gut microbiota, inhibiting TLR4/MyD88/NF-κB signaling pathway and restoring neurotransmitter homeostasis

Chinese Traditional and Herbal Drugs. 2026, 57(7): 2583-2601.

Objective To explore the therapeutic effect and mechanism of Shouhui Tongbian Capsule (首荟通便胶囊, SHTB) on post-stroke depression (PSD) using a multi-omics strategy. Methods A mouse model of PSD was established by photochemical induction of stroke combined with chronic unpredictable mild stress (CUMS). Depressive-like behaviors were evaluated by behavioral tests, and neuronal and myelin sheath structures were assessed by histological analysis. A comprehensive approach integrating 16S rDNA sequencing of gut microbiota, metabolomics of feces and brain tissues, proteomics, neurotransmitter detection and Western blotting was employed to systematically elucidate the potential mechanism. Additionally, gut microbiota was depleted by antibiotics to verify whether the effect of SHTB is dependent on intestinal microbes. Results SHTB significantly improved depressive-like behaviors and neuronal morphology in PSD mice. 16S rDNA sequencing analysis showed that SHTB restored the balance of gut microbiota, characterized by an increase in beneficial genera (e.g., Enterococcus) and a decrease in pro-inflammatory genera (e.g., Oscillibacter, Erysipelotrichaceae). Metabolomic results indicated that SHTB could correct fatty acid metabolism disorders, particularly linoleic acid and arachidonic acid pathways, and reduce the levels of neuroinflammation-related metabolites. Proteomics and neurotransmitter detection further revealed that SHTB could restore the levels of serotonin (5-HT), γ-aminobutyric acid (GABA), dopamine (DA) and norepinephrine (NE) (P < 0.05), reduce the abnormal accumulation of glutamate (P < 0.05), and inhibit the excessive activation of TLR4/MyD88/NF-κB inflammatory pathway (P < 0.05). After depletion of gut microbiota by antibiotics, the above effects were significantly attenuated, indicating that the efficacy of SHTB was gut microbiota-dependent. Conclusion SHTB could restore the gut microbiota, repair the intestinal barrier, regulate fatty acid metabolism, inhibit TLR4/MyD88/NF-κB pathway-mediated peripheral and central inflammatory responses, and restore the balance of key neurotransmitters through a gut microbiota-dependent mechanism. This multi-dimensional improvement in the gut-brain axis function contributes to its anti-PSD and neuroprotective effects.

Mechanism of Zuogui Jiangtang Jieyu Formula in improving synaptic loss of hippocampal neurons via regulating microglial TREM2/C1q signaling in diabetes-related depression rats

Chinese Traditional and Herbal Drugs. 2026, 57(7): 2602-2613.

Objective To reveal the mechanism of Zuogui Jiangtang Jieyu Formula (左归降糖解郁方, ZGF) to improve synaptic loss of hippocampal neuron based on triggering receptor expressed on myeloid cells 2 (TREM2)/complement component 1q (C1q) signaling-mediated microglial cell synaptic pruning in diabetes-related depression (DD) rats. Methods The model of DD rats was established and rats were randomly divided into control group, model group, TREM2 inhibitor (IN1, 1.5 μg/kg) group, TREM2 agonist (AL002, 2 μg/kg) group, positive drug (metformin 0.18 g/kg+ fluoxetine 1.8 mg/kg) group and ZGF (10.26 g/kg) group. Primary microglia and hippocampal neurons from SD rats were cultured separately. An in vitro co-culture cell model simulating the DD environment was constructed by treating with 150 mmol/L high glucose combined with 200 μmol/L corticosterone, control group, model group, TREM2 inhibitor (IN1, 2 μmol/L) group, TREM2 agonist (AL002, 5 μmol/L) group, positive drug (10% positive drug-containing serum) group and ZGF (10% ZGF-containing serum) group were set up. Depression-like behavior was evaluated by open field, forced swimming and sucrose preference test. Synaptic loss of hippocampal neuron was observed by Golgi staining. Morphology and structure of microglia and hippocampal neuron was observed by cell imaging analysis. Nissl’s staining was used to observe synaptic loss of hippocampal neuron. The protein expressions of TREM2 and C1q in rat hippocampal microglia or in vitro microglia, and ostsynaptic density protein 95 (PSD95) in rat hippocampal neurons or in vitro hippocampal neurons were detected by immunofluorescence. The expressions of TREM2 protein in hippocampal tissue or in vitro microglia were detected by Western blotting. Results The animal experiment results showed that ZGF significantly increased the total activity distance and sugar water preference rate of rats in open field experiment (P < 0.01), reduced forced swimming immobility time (P < 0.01), up-regulated TREM2 expression in hippocampal microglia (P < 0.01), and inhibited C1q/PSD95 signal mediated excessive pruning of microglial synapses, thereby improving synaptic loss in rat hippocampal neurons (P < 0.05, 0.01). The results of cell experiments showed that the serum containing ZGF could significantly up-regulate TREM2 expression and inhibit C1q/PSD95 signal mediated synaptic pruning in microglia, thereby reversing synaptic loss in hippocampal neurons (P < 0.05, 0.01). Conclusion ZGF effectively mitigate the synaptic loss of hippocampal neuron and thereby alleviate depressive behavior in rats, the mechanism of which might be related to the regulation of excessive synaptic pruning of microglia mediated by TREM2/C1q signaling.

Mechanism of Ligustri Lucidi Fructus with its wine-processed product on attenuating Tripterygium wilfordii polycoride-induced reproductive toxicity based on Nrf2-mediated testicular antioxidant defense

Chinese Traditional and Herbal Drugs. 2026, 57(7): 2614-2621.

Objective To explore the differential attenuation effects of Nüzhenzi (Ligustri Lucidi Fructus) and its wine-processed product on Tripterygium wilfordii polycoride (TWP)-induced reproductive toxicity, and to investigate its attenuation mechanism based on nuclear factor erythroid 2-related factor 2 (Nrf2) mediated testicular antioxidant defense. Methods ICR mice were ig TWP (40 mg/kg) for 28 d, and intervention was also given with Ligustri Lucidi Fructus or its wine-processed products. Reagent kits were used to detect the levels of testosterone in serum and oxidative stress-related indicators in testicular tissue. Hematoxylin-eosin (HE) staining was used to observe pathological changes in testicular tissue. Western blotting and qRT-PCR were used to detect the protein and mRNA expressions of Nrf2, heme oxygenase-1 (HO-1) and nicotinamide adenine dinucleotide phosphate quinone dehydrogenase 1 (NQO1) in testicular tissue. Results TWP caused a loose structure of seminiferous tubules in testes of mice, a decrease in level of spermatogenic cells, a significant reduction in number of mature sperm, a significant decrease in testosterone levels (P < 0.01), and a significant increase in the level of malondialdehyde (MDA), a lipid peroxidation product in testes (P < 0.01). Ligustri Lucidi Fructus (1, 2 g/kg) could significantly inhibit the abnormal indicators mentioned above (P < 0.05, 0.01), and Ligustri Lucidi Fructus (2 g/kg) could also significantly enhance the activity of antioxidant enzymes (P < 0.05, 0.01). Ligustri Lucidi Fructus wine-processed products (1, 2 g/kg) could significantly inhibit the abnormal indicators mentioned above (P < 0.01), and Ligustri Lucidi Fructus wine-processed products (2 g/kg) could more comprehensively reverse oxidative stress imbalance (P < 0.01). At the same dose, the detoxification effect of Ligustri Lucidi Fructus wine-processed products was significantly better than that of their raw products (P < 0.05, 0.01). Mechanism analysis revealed that TWP significantly down-regulated the expressions of Nrf2 and its downstream molecules HO-1 and NQO1 in testicular tissue of mice (P < 0.01), while Ligustri Lucidi Fructus and its wine-processed products (1, 2 g/kg) significantly up-regulated the expressions of Nrf2 and its downstream molecules HO-1 and NQO1 (P < 0.05, 0.01), with Ligustri Lucidi Fructus wine-processed products showing a significantly better effect than Ligustri Lucidi Fructus (P < 0.01). Conclusion Ligustri Lucidi Fructus and its wine-processed products could alleviate the testicular reproductive toxicity caused by TWP, and the detoxification effect of wine-processed products is significantly better than that of raw products. The mechanism may be related to enhancing Nrf2 mediated testicular antioxidant defense.

Exploring causal association between gut microbiota and myocardial infarction from perspective of “gut-heart axis” based on theory of “heart-small intestine relationship” and predicting intervention with traditional Chinese medicine

Chinese Traditional and Herbal Drugs. 2026, 57(7): 2622-2639.

Objective Based on the theory of “heart-small intestine relationship” and the “gut-heart axis” hypothesis, this study applies Mendelian randomization (MR) and bioinformatics approaches to investigate gut microbiota with genetic causal effects on myocardial infarction (MI), explore underlying mechanisms, and predict potential interventional traditional Chinese medicines. Methods Genome-wide association study (GWAS) and gene expression data for gut microbiota and MI were obtained from the MiBioGen, IEU GWAS and GEO databases. MR analysis was primarily performed using the inverse variance weighted (IVW) method. Potential mechanisms were explored through gene ontology (GO) and Kyoto encyclopedia of genes and genomes (KEGG) enrichment analyses. Core genes were screened via machine learning algorithms. Potential traditional Chinese medicines were predicted by integrating the Coremine database, and their properties (four qi), flavors (five tastes), meridian tropism, and functions were statistically analyzed. Results MR analysis revealed six gut microbiota taxa with genetically causal associations with MI: Butyricicoccus (a protective factor, OR < 1, P < 0.05), Lachnoclostridium, Ruminococcaceae UCG014, Odoribacter, Oxalobacter, and Sellimonas (risk factors, OR > 1, P < 0.05).The core genes were mainly enriched in signaling pathways including the ephrin receptor signaling pathway, calcium signaling pathway, and mitogen activated protein kinases (MAPK) signaling pathway. A total of 596 potential traditional Chinese medicines were predicted. Their properties (four qi) were predominantly warm, followed by cold and neutral; their flavors (five tastes) were primarily sweet, with bitter and pungent also being notable. The main therapeutic functions identified were clearing heat and detoxifying. Representative medicines included Renshen (Ginseng Radix et Rhizoma), Sanqi (Notoginseng Radix et Rhizoma), and Huanglian (Coptidis Rhizoma). Related classical prescriptions encompassed Xiaochaihu Decoction (小柴胡汤) and Taohong Siwu Decoction (桃红四物汤). Conclusion This study reveals the genetic causal relationship and molecular mechanisms through which gut microbiota mediates the occurrence of MI. It enriches the modern biological connotation of the “heart-small intestine relationship” theory. The predicted traditional Chinese medicines provide new strategies for the traditional Chinese medicine-based prevention and treatment of MI and offer a reference for further investigation into the “gut-heart axis”.

Analysis of core mechanisms of ischemic stroke based on machine learning and experimental verification and prediction of traditional Chinese medicine prevention and treatment strategies

Chinese Traditional and Herbal Drugs. 2026, 57(7): 2640-2654.

Objective To screen key genes of ischemic stroke (IS) and analyze their mechanisms by integrating multi-omics and machine learning methods, and predict their potential targets for traditional Chinese medicine prevention and treatment. Methods IS transcriptome data from GEO database was integrated, and candidate genes were screened through differential expression, weighted gene co-expression network analysis (WGCNA) and protein interaction network analysis. A variety of machine learning algorithms including logistic least absolute shrinkage and selection operator (Lasso), random forest, etc. were used to construct a diagnostic model, and the optimal gene set is determined through cross-verification. An middle cerebral artery occlusion (MCAO) model was constructed and further verified by Bederson scoring, HE staining and real-time quantitative polymerase chain reaction (RT-qPCR). Immune infiltration was analyzed using CIBERSORTx and potential traditional Chinese medicines were reverse-matched based on Coremine Medical database. Results Eight core genes (ARG1, CLEC4E, CLEC5A, FCAR, FCGR1A, IRAK3, MCEMP1, TLR5) were identified, and their diagnostic models performed well in both the training and verification cohorts [area under curve (AUC) > 0.7]. Animal experiments had confirmed that the expression of these genes was significantly up-regulated in the cortical tissue of IS model rats, and was closely related to the infiltration level of immune cells such as M0-type macrophages and neutrophils. Based on the above targets, 59 potential traditional Chinese medicines were predicted. Most of the four qi were cold, warm and calm, and most of the five are bitter and sweet. The meridian tropism was mainly concentrated in the liver, kidney and spleen meridians. The medicinal properties and meridian tropism were consistent with the pathogenesis of IS “liver and kidney yin deficiency, and stasis and heat accumulation”. Conclusion By integrating bioinformatics, machine learning and experimental validation, this study systematically identified and validated eight key genes involved in the remodelling of the post-stroke immune microenvironment, which may serve as potential diagnostic biomarkers for IS. Predictive analysis of traditional Chinese medicine suggests that the pharmacological properties—including taste, nature and meridian tropism—of drugs targeting these genes are consistent with the pathogenesis of IS, thereby providing a theoretical basis for the prevention and treatment of IS using traditional Chinese medicine from the perspective of immune regulation.

Analysis of knowledge graph, global patents and trend prospect of Eleutherococcus plants

WEI Xinyue, LI Siya, HUA Haibing, HE Runran, XI Feifei, WANG Jin, ZHANG Yufeng

Chinese Traditional and Herbal Drugs. 2026, 57(7): 2655-2673.

Objective To conduct a bibliometric and visual analysis using CiteSpace and VOSviewer on the current research, status of plants in the genus Eleutherococcus and predict future research trends. Additionally, by analyzing the patent landscape of Eleutherococcus plants, this study aims to evaluate the current status and development trends of its technological transformation, thereby providing a decision-making basis for subsequent research, development, and industrialization. Methods Literature searches were performed in four databases: China National Knowledge Infrastructure (CNKI), Chinese Sci-Tech Journal Full-text Database (VIP), Wanfang Data Knowledge Service Platform (Wanfang), and Web of Science (WOS), using keywords such as “Eleutherococcus” The retrieved literature was imported into NoteExpress software for screening and deduplication. The exported data were then visualized using CiteSpace and VOSviewer to analyze the distribution of the number of published papers, author and institutional collaboration networks, as well as keyword clustering and co-occurrence networks. The search function of the Incopat patent database was utilized to analyze and summarize global patent trends, geographical distribution, applicant rankings, and applicant type composition related to Eleutherococcus plants. Results A total of 2 081 Chinese articles and 462 English articles meeting the criteria were included. Over the past decade, the number of Chinese publications on Eleutherococcus generally showed a fluctuating decline before stabilizing, while English publications exhibited a fluctuating upward trend. China was the leading country in terms of publication volume. Author collaboration networks were primarily based on intra-team cooperation, with relatively insufficient inter-team collaboration. Through the Incopat patent database, 18 992 patent records related to Eleutherococcus plants were retrieved. Among these, 9 942 patents (52.35%) were domestic applications in China, accounting for more than half of the global patents in this field. Conclusion Domestic research on Eleutherococcus in China is currently in a phase of fluctuating decline, while international research is on the rise. Studies predominantly focus on Acanthopanax senticosus and A. sessiliflorus, with research hotspots trending toward active components, oxidative stress, modern pharmacology, and resource conservation. The global distribution of patents related to Eleutherococcus plants shows significant disparities. Although China has a relatively high number of patent applications, it faces challenges such as reduced application activity in recent years and insufficient innovation.

Scientometrics and visualization analysis of Gardeniae Fructus

Chinese Traditional and Herbal Drugs. 2026, 57(7): 2674-2690.

Objective This paper uses scientometrics methods to conduct a visual analysis of Chinese and English literature on the Zhizi (Gardeniae Fructus) to explore its research progress and future development trends. Methods Bibliometrix and VOSviewer software were used to conduct a scientometric analysis of research literature in the field of the Gardeniae Fructus in the China National Knowledge Infrastructure (CNKI), VIP, Wanfang, Web of Science, Scopus, PubMed databases up to 2025. From the perspectives of annual publication volume, countries, institutions, journals, keywords, and theme evolution, a visual analysis of the research on Gardeniae Fructus at home and abroad was conducted. Results A total of 5 049 relevant Chinese and 639 English literature were retrieved. The countries that conduct research in the field of Gardeniae Fructus are mainly China. The research forces show a significant regional and institutional concentration feature, and the core contributions are mostly from leading Chinese universities of traditional Chinese medicine. In recent years, the popularity of research on the Gardeniae Fructus has continued to grow, with both Chinese and English literature focusing on the pharmacological mechanisms of active ingredients such as geniposide, extraction process optimization, and quality control system construction. Chinese research focuses more on the application and industrial development of traditional formulas, while English research focuses more on molecular mechanisms and international collaboration. Scientometric analysis indicates that the pharmacological activity and toxicity of key active ingredients in the Gardeniae Fructus, as well as research methods such as network pharmacology, metabolomics, molecular docking, and response surface methodology, are currently the focus and hotspots of research. Conclusion This study analyzed and visualized the Chinese and English literature on the Gardeniae Fructus using scientometric methods, and identified the research priorities and hot topics in the pharmacological activities, toxic effects, and research techniques and methods of Gardeniae Fructus. This has certain reference significance for in-depth research on the Gardeniae Fructus and its active ingredients, as well as for the selection of future research topics.

Identification of GLR gene family in Panax notoginseng and their expression analysis under different hormonal treatments

LI Junliang, YANG Zihan, LYU Wenhui, LIU Guanze, YANG Shengchao

Chinese Traditional and Herbal Drugs. 2026, 57(7): 2691-2703.

Objective The glutamate receptor-like (GLR) gene family was identified at the whole genome level of Panax notoginseng and the expression analysis under different hormone treatments was conducted to provide a reference for in-depth study of its functions. Methods The PnGLR gene family was systematically identified using bioinformatics methods, followed by comprehensive analyses of physicochemical properties, phylogenetic relationships, conserved motifs, protein structures, cis-regulatory elements, and expression patterns. Additionally, their expression responses were examined under exogenous hormone treatments, including methyl jasmonate (MeJA), salicylic acid (SA), abscisic acid (ABA), and L-glutamic acid (L-Glu), as well as upon infection with Cylindrocarpon destructans. Results Genome-wide analysis identified 12 PnGLR genes in P. notoginseng, which were unevenly distributed across six chromosomes and grouped into four clades. Members of the PnGLR gene family encoded 693 to 1372 amino acids and contained 2 to 5 transmembrane domains, and were all predicted to localize to the plasma membrane. The cis-acting element prediction results showed that the promoter region of PnGLR contained response elements related to light, plant hormones, and abiotic/biotic stress. Real-time quantitative polymerase chain reaction (RT-qPCR) results demonstrated that PnGLR2, PnGLR4, PnGLR5, PnGLR8, PnGLR9, and PnGLR11 were significantly upregulated under exogenous hormone treatments and C. destructans infection. Exogenous L-Glu treatment with 1 and 10 mmol/L could significantly induce the upregulation of gene expression of PnGLR5, PnGLR6, PnGLR7, PnGLR9, and PnGLR10. Conclusion A total of 12 members of the PnGLR gene family were identified at the whole-genome level, and their expression patterns under exogenous hormone, L-Glu treatment and C. destructans infection were different, this laid a theoretical foundation for further exploring the potential functions of the PnGLR genes.

Identification of R2R3-MYB transcription factor family of Bupleurum chinense and expression analysis under salt stress

Chinese Traditional and Herbal Drugs. 2026, 57(7): 2704-2717.

Objective To identify and analyze the expression of R2R3-MYB transcription factor family of Bupleurum chinense, and provide a theoretical basis for analyzing the secondary metabolic regulation mechanism of B. chinense under salt stress. Methods All R2R3-MYB genes of B. chinense were screened and identified by homologous alignment and hidden Markov model. Their physical and chemical properties, phylogenetic relationship, conserved domains, gene structure, cis-acting elements and chromosome distribution were systematically analyzed, and their expression patterns under 200 mmol/L salt stress were analyzed by transcriptome sequencing. Results A total of 116 R2R3-MYB transcription factors were identified in the B. chinense genome, all of which were hydrophilic nuclear localization proteins, with only two members containing transmembrane domains. Phylogenetic analysis showed that they clustered into 23 subfamilies, mainly distributed in K, P, W, and X subfamilies. The promoters contained four types of cis-acting elements, including light response, hormone response, stress response, and development regulation, among which light response elements accounted for the highest proportion. There were 42 pairs of gene duplication events, including three tandem duplications and 39 fragment duplications. The salt stress expression profile revealed that 21 genes responded significantly, and they were classified into early response type, mid-term continuous type, and late inhibition type according to the expression patterns. Conclusion This study conducted a genome-wide identification, evolutionary analysis and salt stress expression analysis of the R2R3-MYB gene family in B. chinense. It was initially determined that BcMYB59 and BcMYB116, due to their unique structural features and strong stress response, are excellent candidate genes for understanding the salt stress response mechanism and potential transmembrane signal perception function of B. chinense.

Gene cloning and expression analysis of cytochrome P450 family gene CaCYP2 from Centella asiatica

LIU Huanying, LEI Ming, XIAO Xiao, LI Linxuan, LI Cui, HUANG Yuan, HUO Juan, WAN Lingyun, PAN Limei, ZHANG Zhanjiang

Chinese Traditional and Herbal Drugs. 2026, 57(7): 2718-2729.

Objective To clone the cytochrome P450 gene (CaCYP2) involved in C-6β hydroxylation modification of triterpenoid saponins in Centella asiatica, systematically analyze its molecular characteristics, tissue expression pattern and subcellular localization, and achieve prokaryotic soluble expression, so as to provide experimental basis for clarifying its function in triterpenoid biosynthesis. Methods The full-length sequence of CaCYP2 was cloned from C. asiatica cDNA and subjected to bioinformatics analysis. Quantitative reverse transcription polymerase chain reaction (qRT-PCR) was performed to detect the gene specific expression in different tissues. Seamless cloning technology was employed to construct yeast two-hybrid vector, prokaryotic expression vector and subcellular localization vector for self-activation detection, recombinant protein expression and tobacco transient localization analysis, respectively. Results The CaCYP2 gene was successfully cloned. Its open reading frame (ORF) was 1 443 bp, encoding 480 amino acids, and it belonged to the CYP716 family, hence named CYP716E116. The predicted protein had a relative molecular mass of 54 430 and a theoretical isoelectric point of 8.98, showing hydrophilicity. It contained a typical CYP90-like conserved domain and one transmembrane domain. Subcellular localization results indicated that this protein was localized in the endoplasmic reticulum. qRT-PCR analysis showed that the expression level of CaCYP2 was highest in the leaves, which was 6.2 times higher than that in the roots (P < 0.000 1), showing significant differences compared to other tissues. Prokaryotic expression results indicated that a soluble recombinant protein with a relative molecular mass of approximately 91 300 (including a 40 300 tag protein) was obtained after isopropyl β-D-thiogalactoside (IPTG) induction. The yeast two-hybrid experiment confirmed that the yeast two-hybrid vector had no self-activating activity and could be used for subsequent screening of interacting proteins. Conclusion This study successfully cloned the CaCYP2 gene and characterized its molecular structure, tissue-specific expression pattern, and endoplasmic reticulum localization. It also confirmed that the protein can be expressed solubly in a prokaryotic system. These findings lay an important foundation for further investigation into its catalytic function and regulatory role in the biosynthetic pathway of triterpenoid saponins in C. asiatica.