Traditional Medicinal Plant Research Articles

Antitumoral effect thought ROS production of the sesquiterpene lactone centratherin isolated from Eremanthus crotonoides, an endemic plant from Brazil

Cancer is a global public health problem, requiring the development of new and more effective drugs for treatment. Eremanthus crotonoides, is a common plant found in the Brazilian Cerrado and Atlantic Forest and is used as a traditional medicinal plant in some regions. This study evaluated the potential antitumor effect of centratherin, a compound isolated from E. crotonoides leaves, against cancer cells. In colorectal cancer cells, centratherin showed greater toxicity than carboplatin while maintaining comparable selectivity to the control. Although centratherin did not induce classical apoptotic phenotypes and was not inhibited by Zvad, it did induce elevated levels of reactive oxygen species (ROS) that was responsible for cell death explaining its toxicity. in silico analysis, showed that this substance has high oral bioavailability and intestinal absorption. Centratherin was found to be selective against colon cancer cells and, therefore may be considered for future studies on colorectal cancer (CRC) studies.

Comparative transcriptomic analysis provides insights into the regulation of root-specific saponin production in Panax japonicus

Panax japonicus, a traditional medicinal plant from the Araliaceae family, produces bioactive triterpenes with health benefits. In Traditional Chinese Medicine, its roots have been used, but the chemical basis of its medicinal use is unclear, particularly regarding the metabolism and regulation of triterpene saponin biosynthesis. This study employed an integrative approach using Ultra Performance Liquid Chromatography (UPLC) and transcriptome analysis. Our UPLC analysis showed that ginsenoside Ro and chikusetsusaponin IVa were mainly detected in P. japonicus root. Subsequently, a comparative transcriptome analysis of four P. japonicus tissues (roots, leaves, flowers and fruits) was conducted using Illumina sequencing. As a result, 90,985 unigenes were functionally annotated from a total of 211,650 assembled non-redundant transcripts. Among these, 42,829 unigenes were annotated in NR database. Tissue-specific gene analysis revealed that roots had the highest number of specifically expressed unigenes (11,832). The majority of these unigenes were associated with metabolic processes. Additionally, tissue expression patterns analysis for three common transcription factor families indicated that WRKY family genes showed a significantly root-specific expression pattern, potentially playing a role in triterpene saponin biosynthesis. Notably, we investigated the expression profiles of genes related to the biosynthesis of triterpene saponins and found that four genes, ACCT, HMGS, HMGR and SS, encoding key enzymes in triterpene saponins biosynthesis pathway, were primarily expressed in the root. Overall, our study provides a set of P. japonicus tissue transcriptome data, which will aid in the discovery of triterpene saponin biosynthetic genes and offers valuable genetic information for this medical plant.

Evaluation of Sedative activity of Cannabis Sativa in mice

Background: Cannabis sativa Linn. (Family: Cannabaceae) is commonly cultivated in Bangladesh in moist areas such as riverbanks, lakes, canals, and roadsides. The leaves of this plant are traditionally used to treat insomnia, depression, and neurodegenerative diseases. Objective: The study's objective is to investigate the sedative effects and phytochemical composition of the Cannabis sativa species in Nigeria. Methods: The elevated plus maze was used to assess the sedative effect of various fractions of aqueous methanol extracts of Cannabis sativa at a dosage of 100 mg/kg in mice. Comparisons were made between the sedative action of the extract and diazepam, which was used as the positive control. Additionally, acute toxicity was also evaluated during the assessment. Results: During the elevated plus-maze test, various parameters such as the number of entries in open arms and the number of entries in closed arms were recorded. The administration of different fractions of aqueous methanol extracts of Cannabis sativa resulted in a significant increase (p < 0.001) in the time spent in and the number of entries into open arms compared to the control group. Conclusion: The findings suggest that various concentrations of water-methanol extracts from Cannabis sativa exhibit sedative effects that are superior to those of the control (diazepam), supporting the use of this plant in traditional medicine. Further research should be conducted to investigate the active components of Cannabis sativa and understand the mechanism of its effects before proceeding to clinical trials.

Haploid-Phased Chromosomal Telomere-to-Telomere Genome Assembly of Medicinal Plant Uncaria rhynchophylla Dissects Genetic Controls on the Biosynthesis of Bioactive Alkaloids.

Natural indole alkaloids provide important medicinal resources and defences to environmental stresses. The Uncaria genus is a recorded traditional medicinal woody plant with high alkaloids. Genomic insights into alkaloid variation remain elusive. Here, we have dissected the haploid-resolved chromosomal T2T genome assembly of Uncaria rhynchophylla with a size of ~634 Mb and contig N50 of 27 Mb using PacBio HiFi long-reads plus Hi-C reads and anchored the contigs on 22 pairs of confirmed chromosomes. This genome contains 56% repeat sequences and ~29 000 protein-encoding genes. U. rhynchophylla diverged from a common ancestor shared with Coffea around 20 million years ago and contains expanded and contracted gene families associated with secondary metabolites and defences/resistance to stresses. We constructed the pathway and mined genes for rhynchophylline alkaloid biosynthesis. Fifty-three alkaloids in this pathway and eight differentially expressed genes are the keys to alkaloid accumulation. Elevated alkaloid levels are driven by high copy numbers of critical genes STRs and SGRs involved in strictosidine synthesis and hydrolysis as evidenced by phylogenetic, expression and RNA interference analyses. These results advance our genetic understanding and guide further breeding improvements, stress adaptation studies and pharmaceutical development.

A Study on Antioxidant and Gastro-protective Effects of Trachyspermum ammi Linn Extract Using Different In-vitro Techniques

The use of a plant's seeds, berries, roots, leaves, bark, or flowers for therapeutic purposes is known as herbal medicine, often called botanical medicine or phytomedicine. There is a long history of using herbs outside of traditional treatment. The use of herbal medicine in treating and preventing disease is becoming more widespread due to advancements in clinical research, analysis, and quality control. There is a long history of using native plants in traditional medicine. Plants serve as the primary source of resources for the conventional medical procedures practiced in rural and tribal regions in India and other countries. Ajowan, or Trachyspermum ammi, is a widely used name. One of the traditional possible herbs used as a spice in daily life is ajwain, or Trachyspermum ammi L., a member of the Apiaceae family. Fruits, vegetables, herbs, and spices all contain phenolic compounds, which have potent anti-inflammatory, antigenotoxic, antioxidant, and anticancer properties. A Soxhlet extractor was utilized to extract the methanol from the dried seeds of Trachyspermum ammi. The Fenton reaction was used to measure the seed's total phenolic content, and it showed good antioxidant activity at various dosage concentrations. With ascorbic acid serving as the standard reducing agent, the antioxidant activity of the seed extract was evaluated. The UV-visible spectrophotometer was used to obtain the current results. A notable concentration-dependent free radical scavenging and reduction power was demonstrated in this plant extract, Trachyspermum ammi. Using SGOT and SGPT enzymes, we also determined that the gastroprotective action of T. ammi extract was significantly higher than that of the standard medication Liv52 and control liver enzymes. The liver histology slide treated with T. ammi extract shows a typical liver lobule. The hepatocytes are polyhedral, have eosinophilic cytoplasm, and typically have a central nucleus. Endothelially lined sinusoids are located between the hepatocyte cords and exhibit normal liver histology compared to control and Liv52-treated groups. Significantly less intestinal histology was observed as compared to untreated control. The current study concludes that Trachyspermum ammi extract may have a natural antioxidant and gastroprotective activity. Trachyspermum ammi is a great way to add gastroprotective and antioxidant phenolic components to your diet.

Identification of New Compounds from the Roots of Rubia tibetica Hook. f.

Rubia tibetica Hook. f is a traditional Tibetan medicinal plant, with its roots serving as the primary medicinal part. Two new compounds, Rubiaxylm C and E (1 and 3), and one new natural compound, Rubiaxylm D (2) were isolated from R. tibetica, along with five known compounds (4-8). The structures of the previously undescribed compounds were elucidated by HR-ESIMS, 1D/2D NMR spectroscopic data and electronic circular dichroism calculations. Compound 1 is a rare anthrone substituted with a carbonyl group at position 1. Furthermore, all isolated compounds were tested for their cytotoxicity against HepG2 cell lines.

3-Oxo-11αH-germacra-1(10) E,4Z-dien-12,6α-olide, a sesquiterpene from Artemisia sieversiana, attenuates lipopolysaccharide-induced inflammation via NF-κB/MAPK pathways and oxidative stress via ROS pathway in RAW264.7 cells.

Inflammation is a vital and normalphysiological response; however, excessive inflammation can contribute to the development of various diseases. Artemisia sieversiana, a traditional Chinese medicinal plant, contains a variety of chemical compounds. One such compound, 3-oxo-11αH-germacra-1(10)E,4Z-dien-12,6α-olide, a germacranolide sesquiterpenoid (germacranolide, GMO), has not been thoroughly investigated regarding its potential anti-inflammatory properties. In this study, the anti-inflammatory and antioxidant properties of GMO were investigated for the lipopolysaccharide (LPS)-induced inflammation in RAW264.7 cells. It was demonstrated that GMO effectively suppressed the production of inflammatory mediators, decreased the phosphorylation of nuclear factor kappa B (NF-κB) and mitogen-activated protein kinase (MAPK) in RAW264.7 cells. Additionally, GMO exhibited the capacity to mitigate oxidative damage induced by LPS, as indicated by assessments of reactive oxygen species and mitochondrial membrane potential. In summary, GMO possesses significant anti-inflammatory effects by modulating the NF-κB/MAPK pathway and antioxidant effects by regulating ROS production.

Chromosome-scale genome assembly of Codonopsis pilosula and comparative genomic analyses shed light on its genome evolution.

Codonopsis pilosula is a significant plant in traditional Chinese medicine, valued for its edible and medicinal properties. However, the lack of available genomic resources has hindered further research. This study presents the first chromosome-scale genome assembly of C. pilosula using PacBio CLR reads and Hi-C scaffolding technology. Additionally, Ks analysis and syntenic depth analysis were performed to elucidate its evolutionary history. The final assembly yielded a high-quality genome of 679.20 Mb, which was anchored to 8 pseudo-chromosomes with an anchoring rate of 96.5% and a scaffold N50 of 80.50 Mb. The genome assembly showed a high completeness of 97.6% based on Benchmarking with Universal Single-Copy Orthologs (BUSCO) analysis. Repetitive elements constituted approximately 76.8% of the genome, with long terminal repeat retrotransposons (LTRs) accounting for about 39.17%. Ks and syntenic depth analyses revealed that the polyploidization history of three platycodonoid clade species involved only the γ-WGT event. Karyotype evolutionary analysis identified an ancestral karyotype with 9 protochromosomes for the three platycodonoid clade species. Moreover, non-WGD genes, particularly those arising from tandem duplications, were found to contribute significantly to gene family expansion. These findings provide essential insights into the genetic diversity and evolutionary biology of C. pilosula, aiding its conservation and sustainable use.

Functional analysis of (E)-β-farnesene synthases involved in accumulation of (E)-β-farnesene in German chamomile (Matricaria chamomilla L.)

German chamomile (Matricaria chamomilla L.) is a traditional medicinal aromatic plant, and the sesquiterpenoids in its flowers have important medicinal value. The (E)-β-farnesene (EβF) is one of the active sesquiterpenoid components and is also a major component of aphid alarm pheromones. In this study, two EβF synthase (βFS) genes (McβFS1 and McβFS2), were cloned from German chamomile. Subcellular localization analysis showed that both McβFS1 and McβFS2 were localized in the cytoplasm and nucleus. Tissue-specific expression analysis revealed that McβFS1 and McβFS2 were expressed in all flower stages, with the highest levels observed during the tubular flower extension stage. Prokaryotic expression and enzyme activity results showed that McβFS1 and McβFS2 possess catalytic activity. Overexpression of McβFS1 and McβFS2 in the hairy roots of German chamomile led to the accumulation of EβF, demonstrating enzyme activity in vivo. The promoters of McβFS1 and McβFS2 were cloned and analyzed. After treating German chamomile with methyl jasmonate (MeJA) and methyl salicylate (MeSA), the transcription levels of McβFS1 and McβFS2 were found to be regulated by both hormones. In addition, feeding experiments showed that aphid infestation upregulated the expression levels of McβFS1 and McβFS2. Our study provides valuable insights into the biosynthesis of EβF, laying a foundation for further research into its metabolic pathways.

The Role of Traditional Medicinal Plants in Managing Malaria and HIV Co-Infection

Malaria and HIV co-infection presents a significant public health challenge, particularly in Sub-Saharan Africa, where co-morbidity rates are highest. Conventional therapies, including antiretrovirals and antimalarial drugs, are expensive and often associated with side effects that lead to noncompliance. Traditional medicinal plants offer an affordable and culturally accepted alternative. This review explores various plants used historically in the treatment of both malaria and HIV, focusing on their phytochemical constituents, mechanisms of action, and clinical efficacy. Plants such as Catharanthus roseus, Adenia venenata, and Morinda lucida have demonstrated antimalarial and antiviral properties, often through bioactive compounds like alkaloids and flavonoids. Additionally, integrating these traditional medicines with modern healthcare systems faces regulatory challenges, but there is promising potential for synergies in treatment efficacy. Further research, clinical trials, and interdisciplinary collaboration are necessary to validate these medicinal plants and improve the management of malaria and HIV co-infection. Keywords: Traditional medicinal plants, Malaria, HIV co-infection, Phytochemicals, Antimalarial properties.

Inventorization of ethnobotanical use of some medicinal halophytes plants in the Algerian arid zone (Biskra)

Context: The population's use of halophyte plants in traditional medicine is a fundamental component of the exploration of natural remedies. Aims: To analyze a list of halophyte plants and their ethnobotanical characteristics, with a focus on those most commonly used in traditional medicine in the southeast of Algeria (Biskra). Methods: The ethnobotanical surveys were conducted between 2017 and 2019 by interviewing 120 informants using a questionnaire and applying a semi-structured interview methodology. The data was analyzed using ethnobotanical statistical tools, including the informant consensus factor (ICF), citation frequency (CF), use value (UV), family importance value (FIV), relative frequency of citation (RFC), plant part value (PPV), and relative importance (RI), as well as statistics analysis (ANOVA) to compare the use of medicinal plants according to age, gender, and education level. Results: The results of the survey enabled us to identify 12 species distributed across eight different families. The most common family was that of the Amaranthaceae (four species), while the most frequently utilized parts of plants were leaves (PPV = 0.4). Furthermore, decoctions were the most commonly prepared form (33.39%). The highest ICF values were observed for the endocrinological and digestive disease groups (ICF = 0.97). The statistical analysis (ANOVA) revealed a significant difference in the knowledge of the use of halophyte plants in traditional medicine according to age and level of education. Conclusions: The findings of this research suggest that the halophyte flora in our arid study area may offer a promising source for the discovery of new drugs, a prospect that requires further investigation and exploration.

Screening and Identification Quality Markers for Antioxidant Activity ofTetrastigma hemsleyanumfrom Different Origins using Spectrum-effect Correlation Analysis

Background::Tetrastigma hemsleyanum (THL) is a traditional medicinal plant with various biological activities, it has been validated that several THL extracts have strong pharmacological activities. Until now, there is no complete and systematic germplasm identification and quality evaluation standard.Objective::Establishing fingerprint spectra of THL from different sources, evaluating their antioxidant activity and identifying quality markers.Methods::In this paper, establishing fingerprints and identifying quality markers through UPLC-QTOF- MS/MS. The antioxidant capacity of forty-one batches of THL from multiple regions was determined by 1,1-Diphenyl-2-picrylhydrazyl radical (DPPH), 2,2'-Azinobis-(3-ethylbenzthiazoline-6- sulphonate) (ABTS), and Ferric reducing antioxidant power (FRAP) assays.Results::The results indicated that THL methanol extracts showed good antioxidative activity and eight THL samples, including THL aerial parts, showed high antioxidative activity. Besides, Ultra Performance Liquid Chromatography (UPLC) fingerprints of THL from different places were established and chromatographic peaks probably correlated with antioxidant capacity were screened by principal component analysis (PCA) and partial least square regression (PLS) analysis.Conclusion::The chemical constituents of these peaks were identified as possible quality markers for THL quality evaluation. In this study, twelve effective substances were preliminarily determined, such as isovitexin, L-epicatechin, procyanidin B, sitosterol, etc.

Isolation, Purification, and Antioxidant Activity of Polyphenols from Cynanchum auriculatum Royle ex Wight

Cynanchum auriculatum Royle ex Wight (CA) is a traditional medicinal and edible plant in China. This study aimed to isolate and characterize the phenolic compounds of C. auriculatum to identify its main antioxidant constituents. Polyphenols were extracted using an ultrasound-assisted ethanol extraction method, followed by partitioning with ethyl acetate. The ethyl acetate extract was then purified through thin-layer chromatography, silica gel column chromatography, and reverse-phase silica gel column chromatography. Three monomeric compounds—cynandione A (I), 2,5-dihydroxyacetophenone (II), and radix piperacanthone (III)—were identified through their physical and chemical properties, UV and IR spectra, and liquid chromatography–mass spectrometry (LC-MS/MS). Vitamin C (VC) and 2,4-dihydroxyacetophenone were used as controls to evaluate the antioxidant potential of the two most abundant monomers. Antioxidant assays demonstrated that 2,5-dihydroxyacetophenone and cynandione A exhibited strong antioxidant activity at lower concentrations, whereas 2,4-dihydroxyacetophenone showed significantly weaker activity. Furthermore, cynandione A displayed superior cellular antioxidant activity compared to 2,5-dihydroxyacetophenone, indicating its potential as a promising bioactive compound. In conclusion, this study provides valuable insights into the phenolic composition of C. auriculatum and highlights cynandione A as a key antioxidant, paving the way for future research on its therapeutic applications.

Induction and Characteristics of Callus Cultures of the Medicinal Plant Tussilago farfara L.

Tussilago farfara L. is a traditional medicinal plant valued for its potentially health-promoting metabolites. Its herbal raw material has been recognized and used since ancient times and continues to be widely used in traditional medicine. Introducing this plant species to in vitro cultivation is a challenging task, but once the protocol is developed, such cultures can provide an abundant and inexhaustible source of plant material. In this study, we report the successful induction and growth of vigorous T. farfara callus in vitro. Callus induction was achieved on MS solid media with the combination of indole-3-acetic acid (3 mg/L) and benzyl aminopurine (2 mg/L) in darkness, whereas it appeared inefficient under light conditions and in suspension culture. We present a detailed description of callus growth kinetics, morphological analysis, photosynthetic activity, and biochemical parameters (including protein content and photosynthetic pigments) supported by histological studies. Furthermore, we observed the potential for organogenesis and somatic embryogenesis. This method for the in vitro propagation of T. farfara, along with callus culture maintenance, offers a wide range of applications in pharmacy for the production of valuable metabolites. Moreover, it could benefit the environment by reducing the depletion of natural populations of this species and may serve as an alternative strategy for species conservation in light of global warming.

Fructus Arctii Mitigates Depressive Disorder via the Let-7e-Modulated Toll-Like Receptor (TLR) Signaling Pathway.

Depressive disorder is a common and serious public health challenge globally. Fructus arctii is a traditional medicinal plant ingredient with diverse pharmacological effects. This study aimed to investigate the therapeutic potential of Fructus arctii in alleviating depressive-like behaviors. We established a chronic unpredictable mild stress (CUMS)-induced depression mouse model to assess the antidepressant effects of Fructus arctii. BV2 cells treated with lipopolysaccharide (LPS) were used to mimic neuronal damage. Behavioral tests, including the sucrose preference test, tail-suspension test, and forced swim test, were conducted to evaluate the impact of Fructus arctii on depressive-like behaviors. Let-7e expression was detected by RT-qPCR, and TLR4 signaling pathway activation was evaluated by western blot analysis, which also assessed the inflammatory response by measuring levels of IL-6, IL-1β, MCP-1, TNF-α, and iNOS. Immunohistological analysis was conducted to detect the expression of microglia markers. Luciferase reporter assays verified the interaction between let-7e and TLR4. Fructus arctii administration effectively alleviated depressive-like behaviors induced by CUMS in mice, as evidenced by improved sucrose preference and reduced immobility time in behavioral tests. Mechanistically, Fructus arctii reversed the CUMS-induced downregulation of let-7e and upregulation of TLR4 and MyD88 protein levels in mice hippocampus tissues. In addition, Fructus arctii suppressed microglial activation and reduced the levels of inflammatory factors by upregulating let-7e. Let-7e was verified to bind to TLR4, thereby negatively regulating its expression. TLR4 overexpression reversed the suppressive effect of let-7e upregulation on inflammatory reactions and microglial activation. Furthermore, intracerebroventricular injection of let-7e agomiR alleviated depressive-like behavior and inhibited microglial activation in vivo. In summary, Fructus arctii mitigates depression by regulating the let-7e/TLR4/MyD88 pathway, offering new insights into potential depression therapies.

Application of DNA Barcoding to Identify Medicinal Plants.

Medicinal plants are valuable resources globally and are used worldwide to maintain health and treat disease; however, the presence of adulteration obstructs their development. DNA barcoding, a technique for species identification by standard DNA regions, facilitates prompt and accurate identification of traditional medicinal plants. The process of DNA barcoding entails six basic steps: 1) processing the medicinal plants, 2) extracting high-quality total DNA from the medicinal plants using centrifugal column method, 3) amplifying target DNA region internal transcribed spacer 2 (ITS2) with universal primers of plants and performing Sanger sequencing, 4) splicing and aligning sequence to obtain the target sequence, 5) matching the barcode sequence against the barcode library for identification, 6) aligning sequence, comparing intraspecific and interspecific variation, constructing phylogenetic neighbor-joining tree. As shown in the results, the universal primer can amplify the target region. Basic Local Alignment Search Tool (BLAST) demonstrates the percentage identified was 100%, and the neighbor-joining tree demonstrates that the splicing sequences were clustered with the A. sinensis OR879715.1 clade, and the clade support value is 100. This protocol provides a reference for applying DNA barcoding technology as an effective method to identify medicinal plants and adulterants.

Biological Potential and Therapeutic Effectiveness of Artemetin from Traditional to Modern Medicine: An Update on Pharmacological Activities and Analytical Aspects.

Plant products derived from natural sources have been used in medicine as a raw material and newer kinds of drug molecules in pharmaceuticals and other allied health sectors. Phytochemicals have numerous medicinal potentials, including anti- ageing, anti-carcinogenic, anti-microbial, anti-oxidant, and anti-inflammatory activity in medicine. Development and biological application of herbal products in modern medicine signified the value of traditional medicinal plants in health care systems. The objective of the present study was to explore the scientific knowledge of the medicinal importance and therapeutic potential of artemetin in medicine. However, scientific investigations for their pharmacological activities in medicine have been done through scientific data analysis of different scientific research work collected from PubMed, Google, Science Direct and Google Scholar in order to know the biological importance of artemetin in medicine. Moreover, analytical data of artemetin have also been discussed in the present work. The present work and scientific data signified the biological potential of artemetin in medicine. Artemetin has been derived from numerous medicinal plants and dietary herbs, including Artemisia absinthium, Artemisia argyi, Achillea millefolium, and Vitex trifolia. Artemetin has anti-malarial, anti-oxidant, anti-apoptotic, anti-microbial, antitumoral, anti-atherosclerotic, anti-inflammatory, hypotensive and hepatoprotective effects. Further, the biological role of artemetin on lipid oxidation, cytokine production, lipoxygenase, and estrogen-like effects was also investigated in the present work. Analytical data on artemetin in the present paper signified their important role in the isolation, separation, and identification of different classes of pure phytochemicals, including artemetin in medicine. Scientific data analysis of artemetin signified its therapeutic potential in medicine for the development of newer scientific approaches for different human disorders.

Learning Traditional Medicinal Plants with Medicinal Properties Website

Learning Traditional Medicinal Plants with Medicinal Properties Website

IN VITRO EVALUATION OF IMMUNOSTIMULANT ACTIVITY OF ISOLATED EMBELIN FROM EMBELIA RIBES

Traditional medicinal plants have long been used in people’s healthcare regimens to treat a wide range of ailments. The ancient medicinal plant Embelia ribes Burm. (E. ribes) has potential for a wide range of pharmacological activities that are linked to embelin, a natural dihydroxy benzoquinone. E. ribes is reported to have immunomodulatory activity, considering which was the foundation to sketch an immunomodulatory study of the isolated embelin from it. Analytical assessment confirmed the isolated embelin by FTIR, UV-Visible spectrophotometry and DSC when compared with the standard. Embelin showed immunostimulant activity in the concentration range of 160 µg mL-1 to 240 µg mL-1. The in vitro study on human PMN cells, observations of phagocytosis of Candida albicans, and NBT reduction assay explain that embelin has an affinity towards PMN cells.

Ammopiptanthus nanus (M. Pop.) Cheng f. stem ethanolic extract ameliorates rheumatoid arthritis by inhibiting PI3K/AKT/NF-κB pathway-mediated macrophage infiltration

Ethnopharmacological relevanceAmmopiptanthus nanus (M. Pop.) Cheng f. (A. nanus), a traditional Kirgiz medicinal plant, its stem has shown potential in treating rheumatoid arthritis (RA) in China, either through oral medication or by topical application directly to the affected joints, but its underlying mechanism of action remains unexplored. Aim of the studyThe purpose of this study is to elucidate pharmacological mechanism of A. nanus in ameliorating RA using a comprehensive approach that combines network pharmacology, molecular docking and experimental evaluations. Materials and methodsFirstly, the major constituents of A. nanus stem ethanolic extract were identified and quantified by High-Performance Liquid Chromatography (HPLC). Disease target data from Gene Cards database was then used to define RA-associated targets. A protein-protein interaction (PPI) network was created via STRING database. The DAVID database powered gene ontology (GO) function and kyoto encyclopedia of genes and genomes (KEGG) pathway enrichment analysis to gain functional insights. In vitro, RAW264.7 cells were treated with A. nanus to investigate the roles of target proteins and pathways during lipopolysaccharide (LPS) - induced inflammation. Immunofluorescence assays were performed to assess the effects of A. nanus on macrophage infiltration. The key targets and signalling pathways were validated using enzyme-linked immunosorbent assay (ELISA), real-time quantitative polymerase chain reaction (RT-qPCR), molecular docking, immunohistochemical analysis, western blotting and immunofluorescence. Finally, the therapeutic potential of A. nanus in RA was evaluated in a carrageenan-induced rat model. ResultsNetwork analysis identified 31 potential targets of A. nanus associated with RA, including 10 hub targets. KEGG analysis highlighted the involvement of PI3K/AKT signaling pathway. In vivo experiments demonstrated that A. nanus treatment significantly protected against carrageenan-induced inflammatory paw tissue and attenuated macrophage infiltration. Both in vivo and in vitro experiments confirmed that A. nanus significantly downregulated the protein expression of COX-2, iNOS and IL-1β, and inhibited PI3K/AKT/NFκB pathway, which are closely linked to RA. Furthermore, molecular docking and cellular thermal shift assay revealed that licoflavanone showed a strong binding affinity with key targets. ConclusionIn summary, this study provides the first evidence of the potent anti-inflammatory activity of A. nanus in experimental RA. The mechanism of action appears to involve inactivation of the PI3K/AKT/NF-κB pathway-mediated macrophage infiltration. These findings indicate that A. nanus has significant potential as a therapeutic potential agent for RA treatment and offer novel insights for future research and drug development in this field.