Traditional System Of Medicine Research Articles

Exploring the antifungal properties of neem (Azadirachta indica) during: A comprehensive review (2010-2020)

Neem (Azadirachta indica) is a versatile medicinal plant with a long history of use in traditional medicine systems. Among its various therapeutic properties, neem has gained attention for its potent antifungal activity. This paper provides a comprehensive review of the antifungal properties of neem, highlighting its mechanisms of action, efficacy against fungal pathogens, and potential applications in clinical and agricultural settings. By synthesizing findings from a wide range of research studies, this review aims to deepen our understanding of neem's antifungal properties and explore its potential as a natural alternative for combating fungal infections.

Network pharmacology integrated with molecular docking and molecular dynamics simulations to explore the mechanism of Tongxie Yaofang in the treatment of ulcerative colitis.

Tongxie Yaofang (TXYF), a classical traditional Chinese medicine, is commonly used in China to treat ulcerative colitis (UC). The aim of this study was to integrate network pharmacology with molecular docking and molecular dynamics simulations to explore the mechanism of Tongxie Yaofang in the treatment of UC. The traditional Chinese medicine systems pharmacology database was used to retrieve the relevant chemical compositions of the herbs contained in TXYF. The DisGeNET, GeneCards, Online Mendelian Inheritance in Man, and Therapeutic Target Database databases were used to retrieve UC-related targets. To construct protein-protein interaction networks and screen for key targets, gene ontology and Kyoto Encyclopedia of Genes and Genomes analyses of the key targets of TXYF in the treatment of UC were performed using R 4.3.2 software. AutoDock Tools 1.5.7 was used for molecular docking. Molecular dynamics simulations of protein complexes and complexes of proteins with small-molecule ligands and eutectic ligands were carried out with Gromacs 2022 software. Network pharmacology analysis revealed that TXYF could act on UC through multiple targets and pathways. It may exert therapeutic effects mainly through the AGE/RAGE, TOLL, JAK/STAT, and Th17 signaling pathways. The possible targets of TXYF in the treatment of UC could be AKT1, BCL2, EGFR, HMOX1, HSP90AA1, and TGFβ1. Molecular docking analysis revealed that AKT1 had the highest binding energy (-10.55 kcal/mol). Molecular dynamics simulations revealed that the complexes formed by the AKT1 protein and the chemical compounds MOL001910 and MOL00035 had good stability and high binding strength. AKT1 may be the most critical target of TXYF in treating UC, and the key chemical components of TXYF in treating UC may include β-sitosterol (MOL000358) and 11alpha,12alpha-epoxy-3beta-23-dihydroxy-30-norolean-20-en-28,12beta-olide (MOL00 1910). This study revealed that TXYF may exert therapeutic effects on UC through multiple targets, multiple biological functions, and multiple signaling pathways. This study provides a new insight into the pharmacological mechanism of TXYF in treating UC.

Exploration of Active Ingredients, Targets, and Potential Mechanisms of Er-Miao-San in the Treatment of Colorectal Cancer Based on Network Pharmacology and Molecular Docking Technology

Background Er-Miao-San (EMS) is a classic prescription in traditional Chinese medicine (TCM) for the treatment of colorectal cancer (CRC) and has shown promising therapeutic effects in clinical practice. However, the specific components and molecular mechanisms of EMS remain unclear. Purpose The aim of this study was to analyze the effective components and molecular mechanisms of EMS in treating CRC through network pharmacology techniques and experimental validation. Methods The Traditional Chinese Medicine Systems Pharmacology database was used to screen the main active chemical components and targets of the EMS formula. The compound structures were verified using the PubChem database, which is an organic small-molecule bioactivity database. GeneCards and OMIM databases were utilized to predict target genes related to CRC. The Cytoscape 3.8 software was used to construct a “Drug-Active Ingredient-Target-Disease” intersection network. The STRING database was employed to analyze the core target protein–protein interaction network shared by EMS and CRC. The core targets were further subjected to Gene Ontology enrichment analysis and Kyoto Encyclopedia of Genes and Genomes pathway enrichment analysis using the R language. Molecular docking between the core targets and the major active chemical components of EMS was performed using AutoDock software. The impact of the core targets on the prognosis of CRC patients was analyzed using the R language. Subsequently, we validated the potential mechanisms predicted by network pharmacology for the inhibition of CRC cell proliferation by the key proteins in the relevant pathways through CCK8 cell proliferation assays and Western blot experiments. Results Molecular docking results showed good docking affinity between the key active components, such as quercetin and baicalein, of EMS and the core targets. Kaplan–Meier survival analysis demonstrated a close correlation between the core targets and the survival prognosis of CRC patients. Cellular experiments showed that EMS significantly inhibited the proliferation of CRC cells and may promote apoptosis and autophagy of CRC cells by suppressing the expression of key proteins in the PI3K/AKT pathway. Conclusion The TCM formula EMS exerts anti-CRC effects through multiple pathways and targets, improving the prognosis and extending the survival period of CRC patients. This study provides preliminary insights into the effective components and molecular mechanisms of EMS in the treatment of CRC, which were preliminarily validated through molecular docking and experimental approaches.

Effect and Mechanisms of Huangqi-Shanzhuyu in the Treatment of Diabetic Nephropathy based on Network Pharmacology and In Vitro Experiments.

Huangqi-Shanzhuyu (HS), a classic combination of Chinese herbal formulae, has been widely used for the treatment of diabetic nephropathy (DN). However, its pharmacological mechanism of action is still unclear. The active ingredients of HS and their potential targets were identified through the Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform (TCMSP) and the DN-related targets were determined from GeneCards, Online Mendelian Inheritance in Man (OMIM), PharmGkb, and Therapeutic Target Database (TTD). The Cytoscape software was used to construct a herb-disease-target network and screen core genes. STRING was employed to generate a protein-protein interaction (PPI) network. Gene Ontology and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analyses were performed to predict the mechanism of action of HS in DN. Animal experiments and molecular docking were used to verify the potential mechanism. In total, 40 active ingredients and 180 effective targets of HS in DN were identified and 1115 DN-related targets were retrieved. From the PPI network, VEGFA, AKT1, IL6, IL1B, TP53, MMP9, PTGS2, CASP3, EGF and EGFR were identified as core genes. The anti-DN mechanism mainly involved multiple signaling pathways such as AGEs-RAGE. Animal experiments and molecular docking analysis confirmed that HS downregulated the expression of IL-1 and IL-6 via kaempferol-mediated inhibition of JNK1 phosphorylation. HS exhibits a therapeutic effect in DN through its multiple ingredients that act on several targets and multiple signaling pathways, including AGEs-RAGE.

An Overview of Pharmaceutical Applications and In vitro Micropropagation Techniques for Rare and Endangered Plant Species

Many rare and endangered plant species possess valuable secondary metabolites with pharmacological applications. These bioactive compounds are often integral to traditional medicine systems, highlighting the cultural significance of these plants. The health benefits of many medicinal species are not fully validated by contemporary scientific research, and some may be facing extinction due to habitat loss, overharvesting or climate change. This situation highlights the urgent need for effective conservation strategies of the species and sustainable cultivation methods. Micropropagation is a valuable technique for producing large numbers of plants from a single explant, significantly aiding in the conservation and commercial cultivation of rare species. Among the various types of explants, shoot tips and nodal segments have been identified as the most effective explants for micropropagation. These explants can be induced to generate multiple shoots in Murashige and Skoog (MS) medium containing Benzyl aminopurine (BAP). Thidiazuron (TDZ), Kinetin (KIN), or 2,4-Dichlorophenoxyacetic acid (2,4-D) are commonly used in MS medium to promote shoot and root development in both direct and indirect organogenesis processes. Rooting of the plantlets was typically achieved using MS medium either supplemented with Indole-3-butyric acid (IBA) or devoid of auxins, depending on the species and the specific requirements for rooting.

Natural Products in Liver Fibrosis Management: A Five-year Review.

Liver fibrosis, characterized by the overproduction of extracellular matrix proteins within liver tissue, poses a rising global health concern. However, no approved antifibrotic drugs are currently available, highlighting the critical need for understanding the molecular mechanisms of liver fibrosis. This knowledge could not only aid in developing therapies but also enable early intervention, enhance disease prediction, and improve our understanding of the interaction between various underlying conditions and the liver. Notably, natural products used in traditional medicine systems worldwide and demonstrating diverse biochemical and pharmacological activities are increasingly recognized for their potential in treating liver fibrosis. This review aims to comprehensively understand liver fibrosis, emphasizing the molecular mechanisms and advancements in exploring natural products' antifibrotic potential over the past five years. It also acknowledges the challenges in their development and seeks to underscore their potency in enhancing patient prognosis and reducing the global burden of liver disease.

Network pharmacology, molecular docking, and molecular dynamics simulations shed light on the mechanism behind Gynostemma pentaphyllum's efficacy against osteosarcoma.

Osteosarcoma (OS) is the most common type of malignant bone tumor, that poses a serious threat to the lives and health of children and adolescents. Traditional Chinese medicines (TCM) have gained attention for treating OS because of their potent anti-cancer effects and fewer side effects. It is commonly understood that Gynostemma pentaphyllum (Thunb.) Makino (GP) exhibits inhibitory effects on most tumors. However, the knowledge of the systematic mechanisms involved is limited. In this study, the Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform (TCMSP) was searched to screen the effective ingredients and corresponding target genes of GP, and disease target databases were searched to identify relevant targets for OS. Venn analysis was used to visualize overlapping genes, which were further extracted using the protein-protein interaction network. R software was used to conduct gene ontology and Kyoto encyclopedia of genes and genomes pathway enrichment analysis, molecular docking and molecular dynamics simulation further validate the binding efficacy of potential therapeutic targets to compound molecules. In total, 161 and 1981 proteins were identified as target genes of GP and OS, respectively, and 104 overlapping genes were identified. Through analysis of the core subnetwork, 12 hub genes were identified, and Kyoto Encyclopedia of Genes and Genomes pathway enrichment analyses revealed that the PI3K/Akt signaling pathway was the most significant. Molecular docking and molecular dynamics simulations show that a high affinity between quercetin and these targets, especially with the combination of TNF free energy (Δ Gbind) minimum, MM/PBSA and MM/GBSA is 42.85 kcal/mol, respectively, and 45.29 kcal/mol. The active ingredients Rhamnazin and Quercetin in Gypenoylum play a therapeutic role in OS through several key targets and pathways. This study provides ideas and references for further research on drug development.

Studies on the Anti-Inflammatory Effect of Xiaoyao Pills in The Treatment of Postmenopausal Osteoporosis in Mice.

Osteoporosis is a common metabolic disease of elderly and postmenopausal women, with no obvious symptoms during its early stages. In the latter stages of this condition, the patients are prone to fractures, and this can seriously affect their health and quality of life. The worldwide increase in life expectancy has made osteoporosis a global concern. The Xiaoyao pills were previously used in the treatment of depression. In addition, the drug appeared to have estrogen-like activity, which affected the expression of ALP, an early osteoblast-specific marker, and COL-1, a major component of bone extracellular matrix. Xiaoyao pills were assessed for their effects on postmenopausal osteoporosis (PMOM) in mice. The target information of each herbal component of Xiaoyao pills was accessed through the Traditional Chinese Medicine Systems Pharmacology (TCMSP) database. Information from GeneCards, OMIM, PharmGkb, TTD, DrugBank, and other websites was used to construct the regulatory network of the herbal complex through Cytoscape and String network to assess the protein interactions. Mice were ovariectomized, and treated with high and low doses of Xiaoyao pills and these were compared to controls. Their symptoms were assessed by immunocytochemistry of bone tissues. The results suggested that Xiaoyao pills had the ability to alleviate the symptoms of PMOM in ovariectomized mice through the IL-17 signaling pathway. This drug has the potential to become a novel therapeutic agent for the treatment of osteoporosis.

Analysis of core functional components in Yinchenhao Decoction and their pathways for treating liver fibrosis

To analyze the core functional component groups (CFCG) in Yinchenhao Decoction (YCHD) and their possible pathways for treating hepatic fibrosis based on network pharmacology. PPI data were extracted from DisGeNET, Genecards, CMGRN and PTHGRN to construct a weighted network using Cytoscape 3.9.1. The data of the chemical components in YCHD were obtained from Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform (TCMSP), and the potential active components and targets were selected using PreADMET Web server and SwissTargetPrediction. A fusion model was constructed to obtain the functional effect space and evaluate the effective proteins to identify the CFCG followed by GO and KEGG pathway enrichment analyses for all the targets. In cultured human hepatic stellate cells (LX-2 cells), the cytotoxicity of different compounds in YCHD was tested using CCK-8 assay; the effects of these compounds on collagen α1 (Col1a1) mRNA expression and the pathways in 20 ng/mL TGF-β1-stimulated cells were analyzed using RT-qPCR and Western blotting. A total of 1005 pathogenic genes, 226 potential active components and 1529 potential targets in YCHD and 52 potential targets of CFCG were obtained. Benzyl acetate, vanillic acid, clorius, polydatin, lauric acid and ferulic acid were selected for CCK-8 verification, and they all showed minimal cytotoxicity below the concentration of 200 μmol/L. Clorius, polydatin, lauric acid and ferulic acid all effectively inhibited TGF-β1-induced LX-2 cell activation. At the concentration of 200 μmol/L, all these 4 components inhibited PI3K, p-PI3K, AKT, p-AKT, ERK, p-ERK, P38 MAPK and p-P38 MAPK expressions in TGF-β1-induced LX-2 cells. The therapeutic effect of YCHD on hepatic fibrosis is probably mediated by its core functional components including benzyl acetate, vanillic acid, clorius, polydatin, lauric acid and ferulic acid, which inhibit the PI3K-AKT and MAPK pathways in hepatic stellate cells.

Exploration of novel human neutrophil elastase inhibitors from natural compounds: Virtual screening, in vitro, molecular dynamics simulation and in vivo study

BackgroundHuman neutrophil elastase (HNE) is an important contributor to lung diseases such as acute lung injury (ALI) or acute respiratory distress syndrome. Therefore, this study aimed to identify natural HNE inhibitors with anti-inflammatory activity through machine learning algorithms, in vitro assays, molecular dynamic simulation, and an in vivo ALI assay. MethodsBased on the optimized Discovery Studio two-dimensional molecular descriptors, combined with different molecular fingerprints, six machine learning models were established using the Naïve Bayesian (NB) method to identify HNE inhibitors. Subsequently, the optimal model was utilized to screen 6925 drug-like compounds obtained from the Traditional Chinese Medicine Systems Pharmacy Database and Analysis Platform (TCMSP), followed by ADMET analysis. Finally, 10 compounds with reported anti-inflammatory activity were selected to determine their inhibitory activities against HNE in vitro, and the compounds with the best activity were selected for a 100 ns molecular dynamics simulation and its anti-inflammatory effect was evaluated using Poly (I:C)-induced ALI model. ResultsThe evaluation of the in vitro HNE inhibition efficiency of the 10 selected compounds showed that the flavonoid tricetin had the strongest inhibitory effect on HNE. The molecular dynamics simulation indicated that the binding of tricetin to HNE was relatively stable throughout the simulation. Importantly, in vivo experiments indicated that tricetin treatment substantially improved the Poly (I:C)-induced ALI. ConclusionThe proposed NB model was proved valuable for exploring novel HNE inhibitors, and natural tricetin was screened out as a novel HNE inhibitor, which was confirmed by in vitro and in vivo assays for its inhibitory activities.

Phytochemical Marker-based Analysis, Antioxidant Potential, and Shelf-Life Study of Geographically Different Samples of Tinospora cordifolia (Stem)

Background: Tinospora cordifolia (TC), commonly referred to as Guduchi, holds a prominent position in the Indian traditional system of medicine and has been cited in Ayurvedic literature for its efficacy in the treatment of a range of ailments, including jaundice, urinary issues, diabetes, prolonged diarrhea, skin conditions, stomachaches, and dysentery. Methods: The present study analyzed the content of phytochemical markers present in the stems of TC collected from five distinct geographical locations (TC-01 to TC-05) and assessed the corresponding variations in their in-vitro antioxidant potential. Results: Lupeol content in various samples of TC varied from 217.9 to 510.9 μg/mg, while palmatine ranged from 0.002% to 0.025%. Total phenolic content (TPC) ranged from 0.77 to 6.35 mg GAE/g wt., and total flavonoid content (TFC) from 12.15 to 20.03 mg QCE/g wt., with IC50 values between 21.02 and 81.81 mg/mL. Elemental analysis of TC-01 to TC-05 showed significant essential elements and permissible levels of heavy metals. A shelf-life study of TC samples indicated stability beyond three years. Conclusion: The considerable diversity in phytochemical content and antioxidant values underscores the substantial influence of geographical variation on the therapeutic potential of TC.

The comprehensive evaluation and mechanism of HOUTTUYNIA CORDATA (Thunb) injection as a complementary therapy for infantile pneumonia

BackgroundHouttuynia cordata injection (HCI), a commercialized product made from the active ingredients extracted from Houttuynia cordata Thunb, has been used clinically in China for >50 years with significant therapeutic effects on pseudorabies herpesvirus infection in vitro and H1N1 influenza virus in mice. Purpose of studyThis work is to evaluate the complementary therapeutic effect of HCI on infantile pneumonia (IP) and to explore its mechanism of action. Study designFirst, the complementary efficacy of HCI in improving IP was evaluated by meta-analysis, with only randomized controlled trials from 5 electronic databases were included in this study. Second, network pharmacology was employed to predict the underlying mechanism of HCI in improving IP. Finally, molecular docking, surface plasmon resonance (SPR) and lipopolysaccharide (LPS)-induced rat IP model were used to verify the predicted results. ResultsMeta-analysis indicated that HCI can complementarily improve pneumonia by increasing cure rate and reducing the duration of fever, cough, and wheezing. Network pharmacology analysis using databases such as Traditional Chinese Medicine Systems Pharmacology (TCMSP), Swiss target prediction, Therapeutic target database and Genecards showed that quercetin, houttuynin and kaempferol in HCI are the main compounds that improve IP and regulate 96 pneumonia-related protein targets. Protein-protein interaction (PPI) network analysis indicated that interleukin-6, tumor necrosis factor α (TNF-α), interleukin-1 β (IL-1β), protein kinase B-1, matrix metalloproteinase 9, interleukin-10, tumor protein 53, epidermal growth factor receptor, C-X-C motif chemokine 8 and interferon gamma are the ten most relevant targets of HCI in improving IP. In the gene ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analysis, 96 targets are involved in 706 GO functions and 148 KEGG pathways. Among them, TNF-α/IL-1β signaling associated with inflammatory response is considered to be an important mechanism in IP. Molecular docking, SPR and LPS-induced IP model confirmed the predicted results. ConclusionsHCI has a complementary therapeutic effect on IP, and its mechanism may be related to the inhibition of TNF-α/IL-1β inflammatory response by two flavonoid compounds.

Acute and subacute oral toxicity evaluation of Ayurvedic formulation Tapyadi loha in rats

Ayurveda is one of the oldest systems of traditional medicine that provides treatments for a wide range of acute and chronic health problems. It is a common myth amongst people that Ayurvedic drugs have no side effects, whereas the fact is that these drugs can cause adverse effects. Despite their wide use, the safety data of many Ayurvedic formulations are still unavailable. Tapyadi loha is an Ayurvedic formulation traditionally claimed for iron deficiency anemia in pregnant and non-pregnant patients. However, no scientific study has been conducted to evaluate its oral toxicity. Hence, the present study evaluated the acute and subacute oral toxicity of the Tapyadi loha according to the OECD test guidelines 425 and 407, respectively. Tapyadi loha did not cause mortality nor any signs of toxicity when given once orally at a dose of 2000 mg/kg. Subacute toxicity study showed no mortality as well as no behavioral, hematological, biochemical and histopathological abnormalities in rats treated with Tapyadi loha formulation at 250, 500 and 1000 mg/kg for 28 days. It is concluded that the Tapyadi loha is safe at a single dose of 2000 mg/kg and 28 days repeated dose of 1000 mg/kg by oral route in rats.

Research and Analysis on the Pathogenesis of Allergic Rhinitis in Children and Its Treatment by Chinese and Western Medicine

Allergic rhinitis in children is a common allergic disease, when it recurs, the symptoms are significant, long-lasting, to children's daily life and learning has brought a huge burden.Western medicine has abundant clinical experience in treating this kind of disease, but the unique dialectical thinking and treatment system of traditional Chinese medicine also show undeniable advantages.The purpose of this study was to explore the etiology of allergic rhinitis in children, the relationship between allergic rhinitis and TCM syndrome types, and the advantages and disadvantages of TCM and WM treatment.It is found that allergen, heredity, environmental factors and immune imbalance are the common causes leading to high incidence of diseases. On the whole, TCM can classify different syndrome types and apply corresponding treatment methods, effective relief of the disease, improve the quality of life.We hope this paper can provide a new perspective and guidance for clinical diagnosis and treatment.

Exploring the pharmacological mechanism of Xianlingubao against diabetic osteoporosis based on network pharmacology and molecular docking: An observational study.

Xianlinggubao formula (XLGB), is a traditional Chinese compound Medicine that has been extensively used in osteoarthritis and aseptic osteonecrosis, but its curative effect on diabetic osteoporosis (DOP) and its pharmacological mechanisms remains not clear. The aim of the present study was to investigate the possible mechanism of drug repurposing of XLGB in DOP therapy. We acquired XLGB active compounds from the traditional Chinese medicine systems pharmacology and traditional Chinese medicines integrated databases and discovered potential targets for these compounds by conducting target fishing using the traditional Chinese medicine systems pharmacology and Swiss Target Prediction databases. Gene Cards and Online Mendelian Inheritance in Man® database were used to identify the DOP targets. Overlapping related targets between XLGB and DOP was selected to build a protein-protein interaction network. Next, the Metascape database was utilized to enrich the Gene Ontology and Kyoto Encyclopedia of Genes and Genomes pathways. In addition, Auto-Dock Vina software was used to verify drug and target binding. In total, 48 hub targets were obtained as the candidate targets responsible for DOP therapy. The anti-DOP effect mediated by XLGB was primarily centralized on the advanced glycation end products (AGEs)-receptor for AGE signaling pathway in diabetic complications and osteoclast differentiation. In addition, AKT serine/threonine kinase 1, tumor necrosis factor, Interleukin-6, vascular endothelial growth factor A and peroxisome proliferator activated receptor gamma, which were considered as potential therapeutic targets. Furthermore, molecular docking results confirm the credibility of the predicted therapeutic targets. This study elucidates that XLGB may through regulating AGEs formation and osteoclast differentiation as well as angiogenesis and adipogenesis against DOP. And this study provides new promising points to find the exact regulatory mechanisms of XLGB mediated anti-DOP effect.

Deciphering the effect of Potentilla fulgens root extract against healthy HUVEC cell line and cancer cell lines (A549 and SKOV-3)

BackgroundPotentilla fulgens, a highly valued indigenous medicinal herb grown in high altitudes of the Himalayan region with anticancer, hypoglycaemic, antibacterial, anti-inflammatory, and antiulcerogenic properties, are used in traditional systems of medicine. This study was aimed to investigate the effect of P. fulgens root extract, as one of the natural alternatives to chemotherapeutic drugs used in cancer treatment, on proliferation, apoptosis, and autophagy of human non-small cell lung cancer cell line (A549), human ovarian cancer cell line (SKOV-3), and healthy human umbilical vein endothelial cell line (HUVEC). MethodsAnti-proliferative effect was assessed by MTT assay. The expression of autophagy and apoptosis-related proteins was evaluated by western blotting. Total oxidant status (TOS) and total antioxidant capacity (TAC) test were determined using standard kit methods. ResultsOur results showed that the extract inhibited proliferation of HUVEC, A549, and SKOV-3 cells in a dose-dependent manner. MTT assay analysis revealed that the extract significantly (P<0.05) induced mortality in HUVEC, A549, and SKOV-3 cells. Western blot results revealed increased expression of NF-κB after the extract treatment but led to the down-regulation in Beclin-1, Bax, extracellular-signal-related kinase 1 and 2, Sequestosome-1, and Cleaved Casp-3 levels. Treatment groups showed an increase in TOS and TAC values in A549 and SKOV-3 cell lines, while HUVEC cell line showed an increase in TAC and a decrease in TOS values, compared to the control group. ConclusionsOur findings indicated that P. fulgens root extract inhibited the proliferation of healthy cells and cancer cells through cell cycle arrest, representing its limited application as therapeutic agent in cancer treatment.

Impact of geographical distribution on polyphenol compostion, antioxidant, antimicrobial and cytotoxic potential of Rhynchostylis retusa (L.) Blume roots

IntroductionRhynchostylis retusa (L.) Blume lies in the heart of majestic Himalayan mountains and is well-documented in Traditional System of medicine. R. retusa roots are used to treat rheumatic disease, asthma, tuberculosis, epilepsy, kidney, and menstrual disorders. Besides earlier reports, current study was focused to investigate locational variations of polyphenols antioxidant, antimicrobial and cytotoxicity activities of R. retusa roots. MethodR. retusa roots of four locations of Himachal Pradesh, India were used in current study. The targeted polyphenols were determined using UPLC-PDA while total phenolic and total flavonoid contents were by spectroscopic method. The antioxidant activity was assessed by DPPH ֹ and ABTS ֗+ assays. Antimicrobial activity was performed against gram-positive (Bacillus subtilis and Staphylococcus aureus) and gram-negative bacteria (Salmonella typhimurium and Escherichia coli) while cytotoxicity activity was against HT29 cancer cell line. ResultIn present study, 50% ethanol revealed higher yield in Kangra region (1375m; 32.1109° N, 76.5363° E) sample followed by water and ethanol. Among all extracts, gallic acid and protocatechuic acid were most abundant phenolic acids. The highest TPC and TFC were found in the ethanol extract of Bilaspur (680m; 31.33°N 76.75°E) sample (155.58±6.92mg GAE/g and 262.25±8.32mg RE/g) and Kangra (135.64±4.36mg GAE/g and 281.75±4.38mg RE/g) regions, respectively. The ethanol extract from Hamirpur (790m 31.6862° N, 76.5213° E), exhibited highest antioxidant activity (IC50 33.31±0.72µg/mL and 12.45±0.68µg/mL), in DPPH ֹ and ABTS ֗+ assays, respectively. The ethanol extract from Bilaspur (5mm and 3mm ZOI for B. subtilis and S. aureus), Kangra (4mm and 2.5mm for B. subtilis and S. aureus), and Hamirpur showed most potent antimicrobial activity. Whereas Mandi (760m; 31.7087° N, 76.9320° E) samples showed least potential as compared to other targeted locations. Further, ethanol extracts of all locations were found most effective in terms of their cytotoxic activity against HT29 cancer cell lines. ConclusionCurrent investigations illuminate ethanol's pivotal role in extracting polyphenols and unleashing the pharmacological potential of R. retusa root extract. Thus, ethanol may be considered as a suitable solvent to extract, fractionate and isolate the bioactive compounds for health perspectives. Findings also suggested that Kangra region (1375m; 32.1109° N, 76.5363° E) sample has highest potential and can be focused for further study.

Potential Mechanisms Underlying the Therapeutic Roles of Gancao fuzi Decoction in Cold-dampness Obstruction Syndrome-type Knee Osteoarthritis.

The key active components and potential molecular mechanism of Gancao Fuzi</i> decoction (GFD) in the treatment of cold-dampness obstruction-type knee osteoarthritis (KOA) remain unclear. To explore the mechanism of GFD in the treatment of cold-dampness obstruction syndrome-type KOA by network pharmacology. The potential active components and targets of the four herbs in GFD (Fuzi, Guizhi, Baizhu, and Gancao) were screened using the Traditional Chinese Medicine Systems Pharmacology (TCMSP) database. The targets of KOA were obtained with the Comparative Toxicogenomics Database (CTD), the GeneCards database, and the DisGeNET database, and the common targets of the drugs and disease were ultimately obtained. Cytoscape (v.3.7.1) was used to draw the active component-target network, and the Search Tool for the Retrieval of Interacting Genes/Proteins (STRING) (v.11.0) database was used to construct the protein interaction network. The Database for Annotation, Visualization, and Integrated Discovery (DAVID) was used for the Gene Ontology (GO) function and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analyses of the intersecting targets. A total of 102 potential active components and 208 targets of GFD in the treatment of cold-dampness obstruction syndrome-type KOA were screened. GFD treatment was found to be closely related to many inflammatory signalling pathways in the treatment of KOA. The effect of GFD on cold-dampness obstruction syndrome-type KOA is mediated by multicomponent, multitarget, and multichannel mechanisms, which provides the basis for further experimental study of its pharmacodynamic material basis and mechanism.

A Network Pharmacology-based Study on the Anti-aging Properties of Traditional Chinese Medicine Sisheng Bulao Elixir.

Traditional Chinese Medicine (TCM) has a rich history of use in preventing senescence for millennia in China. Nonetheless, a systematic method to study the antiaging properties and the underlying molecular mechanism of TCM remains absent. The objective of this study is to decipher the anti-aging targets and mechanisms of Sisheng Bulao Elixir (SBE) using a systematic approach based on a novel aging database and network pharmacology. Bioactive compounds and target proteins in SBE were identified via the Traditional Chinese Medicine System Pharmacology (TCMSP) database. Aging-related proteins were uncovered through alignment with the Ageing Alta database. A compound-target (CT) protein network analysis highlighted key flavonoids targeting aging. Core aging-related proteins were extracted through protein-protein interaction (PPI) network analysis. Molecular docking validated binding activities between core compounds and aging-related proteins. The antioxidant activity of SBE was confirmed using an in vitro senescent cells model. A total of 39 active compounds were extracted from a pool of 639 compounds in SBE. Through a matching process with the Aging Alta, 88 target proteins associated with the aging process were identified. Impressively, 80 out of these 88 proteins were found to be targeted by flavonoids. Subsequently, an analysis using CT methodology highlighted 11 top bioactive flavonoids. Notably, core aging-related proteins, including AKT1, MAPK3, TP53, VEGFA, IL6, and HSP90AA1, emerged through the PPI network analysis. Moreover, three flavonoids, namely quercetin, kaempferol, and luteolin, exhibited interactions with over 100 aging-related proteins. Molecular docking studies were conducted on these flavonoids with their shared three target proteins, namely AKT1, HSP90AA1, and IL6, to assess their binding activities. Finally, the antioxidant properties of SBE were validated using an in vitro model of senescent cells. This study offers novel insights into SBE's anti-aging attributes, providing evidence of its molecular mechanisms. It enhances our understanding of traditional remedies in anti-aging research.

Sanhua Tang protects against ischemic stroke by preventing blood-brain barrier injury: a network pharmacology and experiments.

To assess the effect and mechanism of Sanhua Tang (, SHT) in treating ischemic stroke (IS) through the "Kaitong Xuanfu" theory by using network pharmacology and animal experiments. The active ingredients and targets of SHT and IS were screened by public databases such as Traditional Chinese Medicine systems pharmacology, GeneCards, and online mendelian inheritance in man. Visual network topographies were constructed using R, Cytoscape 3.6.0, AutoDockTools, a user-sponsored molecular visualization system on an open-source foundation, and other software to analyze the correlation between targets and active ingredients. The middle cerebral artery occlusion (MCAO) model was established by operation. Animals were divided into the Sham group, MCAO group (M group), aloe-emodin (AE) group (MCAO rats treated with aloe-emodin), SHT at low dosage (SL group) (MCAO rats treated with SL), SHT at medium dosage (SM group), and SHT at high dosage (SH group). 2,3,5-triphenyl tetrazolium chloride staining was used to detect the volume of cerebral infarction; Nissl staining was used to observe the morphology of neuronal cells; transmission electron microscopy was used to observe the integrity of the blood-brain barrier (BBB); enzyme-linked immunosorbent assay was used to detect the content of interleukin-6 (IL-6), IL-10, tumor necrosis factor α (TNF-α) in serum. Western blot was used to detect the expression of vascular endothelial growth factor A (VEGFA) protein in the cerebral ischemic penumbra. Using network pharmacology and molecular docking validation, four active ingredients (lignan, naringenin, aloe-rhodopsin, and β-sitosterol), seven target proteins (protein kinase b 1, IL-6, TNF, VEGFA, TP53, jun proto-oncogene, and cysteinyl aspartate specific proteinase 3), and inflammatory signaling pathways were identified. Animal experiments showed that the SH and AE groups had fewer neurological deficits, reduced brain infarct volumes, decreased serum inflammatory factor levels, increased expression of VEGFA protein, and less structural damage to neurons and BBB. The present study found that the therapeutic mechanism of SHT against IS may be related to the inhibition of BBB inflammatory damage, which is also the mechanism of "Kaitong Xuanfu." The high-dose group of SHT was relatively effective in regulating inflammatory factors, improving BBB permeability, and protecting neuronal cells from damage.