Efficacy of Citrullus colocynthis seed extract on Earias vittella, Fabricius, (Lepidoptera: Noctuidae): environment sustainable approach / Eficácia do extrato de semente de Citrullus colocynthis em Earias vittella Fabricius (Lepidoptera: Noctuidae): abordagem ambiental sustentável

Abstract Earias vittellaFabricius, 1794 (Noctuidae: Lepidoptera) is deliberated to be one of the most destructive pests of cotton and okra vegetation in the world including Asia. The pest has established resistance to various synthetic insecticides. The use of bio-pesticide is one of the unconventional approaches to develop a vigorous ecosystem without harming non- target pests and beneficial natural insect fauna. In the present study, the toxicity levels of Citrullus colocynthis seed extract have been evaluated against the populations of E. vittellaunder standardized laboratory conditions. The toxic effects of C. colocynthis on development periods, protein contents and esterase activity of the life stages of E. vittella were also evaluated. The toxicity levels of methanol, ethanol, hexane, water and profenofos were evaluated on the 1st instar larvae of E. vittella. LC30 and LC80 concentrations exhibited the effectiveness of methanol-based C. colocynthis seed extract against 1st instar larvae of E. vitella. The enhanced larval and pupal periods were revealed in treated samples during the comparison with untreated samples. The intrinsic rate of increase, net reproductive rate in the LC30 and LC80 concentrations exposed larvae remained less than the control treatment. Fecundity, the esterase activity and protein contents were declined in LC30 and LC80 treated samples as compared to the control. The present findings suggest that C. colosynthis extracts based botanical insecticides are beneficial, ecosystem sustainable and can be integrated with insect management programs from environment safety perspective.

Resumo Earias vittella Fabricius, 1794 (Noctuidae: Lepidoptera) é considerada uma das pragas mais destrutivas de algodão e quiabo no mundo, incluindo a Ásia. Essa praga estabeleceu resistência a vários inseticidas sintéticos. O uso de biopesticidas é uma das abordagens não convencionais para desenvolver um ecossistema saudável sem prejudicar as pragas não alvo e a fauna natural benéfica de insetos. No presente estudo, os níveis de toxicidade do extrato de semente de Citrullus colocynthis foram avaliados nas populações de E. vittella em condições de laboratório padronizadas. Os efeitos tóxicos de C. colocynthis nos períodos de desenvolvimento, conteúdo de proteína e atividade esterase das fases de vida de E. vittella também foram avaliados. Os níveis de toxicidade de metanol, etanol, hexano, água e profenofós foram avaliados em larvas de 1º instar de E. vittella. As concentrações de LC30 e LC80 apresentaram eficácia do extrato de sementes de C. colocynthis à base de metanol contra larvas de 1º instar de E. vittella. Os períodos larval e pupal aumentados foram revelados nas amostras tratadas durante a comparação com as amostras não tratadas. A taxa intrínseca de aumento e a taxa reprodutiva líquida nas concentrações de larvas expostas LC30 e LC80 permaneceram menores do que o tratamento controle. A fecundidade, a atividade da esterase e o conteúdo de proteína diminuíram nas amostras tratadas com LC30 e LC80 em comparação com o controle. As presentes descobertas sugerem que os extratos de C. colocynthis à base de inseticidas botânicos são benéficos, sustentáveis ​​para o ecossistema e podem ser integrados com programas de manejo de insetos do ponto de vista da segurança ambiental.

Antifungal and antibacterial activities of Cannabis sativa L. resins.

ETHNOPHARMACOLOGICAL RELEVANCE: Cannabis sativa L. (Cannabaceae) is a plant native to Eastern Asia spread throughout the world because of its medicinal properties. Despite being used for thousands of years as a palliative therapeutic agent for many pathologies, in many countries research on its effects and properties could only be carried out in recent years, after its legalization. AIMS OF THE STUDY: Increasing resistance to traditional antimicrobial agents demands finding new strategies to fight against microbial infections in medical therapy and agricultural activities. Upon legalization in many countries, Cannabis sativa is gaining attention as a new source of active components, and the evidence for new applications of these compounds is constantly increasing. METHODS: Extracts from five different varieties ofCannabis sativa were performed and their cannabinoids and terpenes profiles were determined by liquid and gas chromatography. Antimicrobial and antifungal activities against Gram (+) and Gram (-) bacteria, yeast and phytopathogen fungus were measured. To analyze a possible action mechanism, cell viability of bacteria and yeast was assessed by propidium iodide stain. RESULTS: Cannabis varieties were grouped into chemotype I and II as a consequence of their cannabidiol (CBD) or tetrahydrocannabinol (THC) content. The terpenes profile was different in quantity and quality among varieties, with (-)b-pinene, b-myrcene, p-cymene and b-caryophyllene being present in all plants. All cannabis varieties were effective to different degree against Gram (+) and Gram (-) bacteria as well as on spore germination and vegetative development of phytopathogenic fungi. These effects were not correlated to the content of major cannabinoids such as CBD or THC, but with the presence of a complex terpenes profile. The effectiveness of the extracts allowed to reduce the necessary doses of a widely used commercial antifungal to prevent the development of fungal spores. CONCLUSION: All the extracts of the analysed cannabis varieties showed antibacterial and antifungal activities. In addition, plants belonging to the same chemotype showed different antimicrobial activity, demonstrating that the classification of cannabis strains based solely on THC and CBD content is not sufficient to justify their biological activities and that other compounds present in the extracts are involved in their action against pathogens. Cannabis extracts act in synergy with chemical fungicides, allowing to reduce its doses.

Antipyretic and antinociceptive effects of methanolic extract of C. iria L. tuber.

ETHNOPHARMACOLOGICAL RELEVANCE: Cyperus iria L. is a sedge belongs to Cyperaceae family. Tuber of this plant is traditionally used in fever. AIM OF THE STUDY: This study aimed to verify the effectiveness of this plant part against fever. Additionally, antinociceptive effect of the plant was evaluated. MATERIALS AND METHODS: Antipyretic effect was evaluated by yeast induced hyperthermia experiment. Antinociceptive effect was determined by acetic acid induced writhing test and hot plate test. Four different doses of plant extract were used in mice model. RESULTS: Extract at dose of 400 mg/kg.bw produced greater effect than paracetamol; reduction of elevated mice body temperature was observed by 2.6 °F and 4.2 °F after 4 h by paracetamol and 400 mg/kg.bw extract respectively. In acetic acid writhing test, extract at 400 mg/kg.bw and diclofenac were found to have equivalent effects producing percentage inhibition of writhing of 67.68% and 68.29% respectively. In hot plat test, significant reduction in latency was also observed after administration of plant extracts. Mean percent maximal effect was 83.55% and 67.26% for ketorolac and extract (400 mg/kg.bw) respectively. CONCLUSION: Our study endorsed the traditional use of C. iria tuber in fever with possible antinociceptive effects.

Dohongsamul-tang inhibits cardiac remodeling and fibrosis through calcineurin/NFAT and TGF-ß/Smad2 signaling in cardiac hypertrophy.

ETHNOPHARMACOLOGICAL RELEVANCE: Dohongsammul-tang (DH) is a Korean traditional herbal medicine used to alleviate symptoms caused by extravasated blood. It is known to protect against cardiovascular diseases and promote blood circulation by activating blood circulation to dispel blood stasis. The DH based on the characteristics of its medicinal properties has discovered the potential of alleviating cardiac hypertrophy. Therefore, this study was performed to verify the pharmacological effect of DH on improving cardiovascular disorders and to demonstrate its mutual improvement effect on renal function. Furthermore, aim of this study is founding the new potential beyond the traditional medicinal efficacy of DH, a traditional medicine. AIM OF THE STUDY: In cardiovascular disease, cardiac hypertrophy refers to a change in the shape of the heart's structure due to pressure overload. It is known that an increase in myofibrils causes thickening of the heart, resulting in high blood pressure. Therefore, suppressing cardiac hypertrophy may be a major factor in lowering the morbidity, mortality, and heart failure associated with cardiovascular disease. Therefore, the study was performed to investigate whether DH, traditionally used, has effects on improving and alleviating cardiac injury and fibrosis caused by cardiac hypertrophy. MATERIALS AND METHODS: Dohongsamul-tang was composed of 6 herbal medicine and each material were boiled with 4 L distilled water for 2 h. The mixture for dohongsamul-tang centrifuged at 3000 rpm for 10 min and concentrated. The concentrated dohongsamul-tang extraction freeze-dried and sotred at 70 °C. The powder of dohongsamul-tang was diluted with distilled water and administered orally. In this study, pressure overload was induced by tying the transverse aortic arch, which is connected to the left ventricle, to the thickness of a 27G needle by performing a surgical operation. The resulting cardiac hypertrophy and heart remodeling was induced and maintained for 8 weeks. RESULTS: The study administered propranolol and dohongsamul-tang orally for 10 weeks to investigate their effects on cardiac hypertrophy induced by transverse aortic contraction (TAC) surgery. Results showed that TAC group increased the left ventricle weight and decreased cardiac function, but dohongsamul-tang treatment attenuated these effects. The pressure-volume curve experiment revealed that dohongsamul-tang improved cardiovascular function, which was worsened by TAC group. Dohongsamul-tang treatment also downregulated collagen I and III through the TGF-ß/Smad2 signaling pathway and improved hematological biomarkers of cardiac hypertrophy. In addition, dohongsamul-tang treatment improved renal function-related biomarkers, such as blood creatinine, blood urea nitrogen, and neutrophil gelatinase-associated lipocalin, which were increased by TAC-induced cardiac hypertrophy. CONCLUSIONS: Taken together, dohongsamul-tang treatment inhibited cardiac remodeling due to pressure overload in the TAC-induced cardiac hypertrophy model, and this effect is thought to be manifested by improving the functional and morphological changes through the calcineurin/NFATc4 and reducing the cardiac fibrosis by suppressing TGF-ß/Smad2 signaling pathways.

Urtica cannabina L. water extract exhibits anti-inflammatory activity by regulating inflammatory cytokines: In vitro and in vivo evidence.

ETHNOPHARMACOLOGICAL RELEVANCE: Urtica cannabina L. (U. cannabina) is a medicinal plant used in traditional Chinese and Kazakh medicine for treatment of various ailments such as rheumatoid arthritis, rheumatic pain, high blood pressure, and snake bites. However, very few studies have focused on the anti-inflammatory effects of U. cannabina and the mechanisms underlying these effects. AIM OF THE STUDY: This study to investigate the in vitro and in vivo anti-inflammatory effect of U. cannabina, the underlying mechanisms, and its phytochemical profile. MATERIALS AND METHODS: We investigated the anti-inflammatory effects of the U. cannabina water extract on lipopolysaccharide-stimulated RAW264.7 macrophages and paw edema in rats and analyzed its chemical components using ultra-performance liquid chromatography-mass spectrometry (UPLC-MS). RESULTS: U. cannabina water extract effectively inhibited the secretion of multiple inflammatory factors, and its corresponding mRNA expression in LPS-induced RAW264.7 cells (p < 0.05). Tincture of U. cannabina water extract significantly reduced carrageenan-induced rat paw edema and levels of inflammatory factors (p < 0.05). A total of 31 compounds, which mainly include organic acids, were tentatively identified based on the comparison of their mass spectrum profiles with those recorded in a mass spectra database. CONCLUSIONS: The results of this study elucidated the anti-inflammatory effect of U. cannabina water extract in vitro and in vivo and showed that the extract elicits the anti-inflammatory effects by regulating the activity of inflammatory cytokines. The results prove that U. cannabina is a valuable source of active compounds with anti-inflammatory activity.

Celastrus orbiculatus Thunb. extract targeting DJ-1 inhibits non-small cell lung cancer invasion and metastasis through mitochondrial-induced ROS accumulation.

ETHNOPHARMACOLOGICAL RELEVANCE: Celastrus orbiculatus Thunb. is an ancient traditional Chinese herb with a long history of medicinal use. The ethyl acetate extract of Celastrus orbiculatus Thunb. (COE) has been shown to have anti-tumor effects in various preclinical studies. However, the anti-invasive and metastatic efficacy of COE in non-small cell lung cancer (NSCLC) and the mechanism by which COE regulates cellular oxidation levels are yet to be elucidated. AIM: To study the anti-dissemination effect of COE on NSCLC and to elucidate the molecular mechanism of COE in regulating cellular oxidation levels and its effect on lung cancer invasion and metastasis. METHODS: CCK-8 assay was used to detect the toxic effects of COE on NSCLC. Transwell assay and high-content imaging was used to detect the Motility of NSCLC. Transmission electron microscopy and three-dimensional (3D) imaging of mitochondrial fluorescence were employed to detect the number and structure of mitochondria. JC-1 probe was used to detect the level of mitochondrial membrane potential. Firefly luciferase assay was used to detect the level of total intracellular ATP. MitoSox probe and DCFH-DA probe were applied to detect the level of reactive oxygen species (ROS) inside the mitochondria and the total intracellular ROS, respectively. Immunohistochemistry was used to detect protein expression in xenograft tumors. RESULTS: COE inhibited motility and induced DJ-1 downregulation in NSCLC at low toxic concentrations, and the antiseptic effect of COE was reduced significantly after the overexpression of DJ-1. COE induced structural disruption of mitochondria in NSCLC and accumulation of superoxide compounds, decreased the volume of membrane potential depolarization, and impaired energy production, ultimately leading to a large accumulation of ROS at the cellular level. The antioxidant acetylcysteine (NAC) significantly reversed the antiseptic capacity of COE. In a xenograft tumor model, protein expression of DJ-1, E-cadherin, N-cadherin, and MMP-2 in COE group was significantly changed compared to the model group. CONCLUSION: In the present study, COE inhibited NSCLC invasion and metastasis and was associated with the downregulation of DJ-1 and elevated ROS. COE-mediated downregulation of DJ-1 may be the primary cause of mitochondrial structural and functional dysfunction in NSCLC, eventually leading to ROS accumulation.

Aqueous M. oleifera leaf extract alleviates DSS-induced colitis in mice through suppression of inflammation.

ETHNOPHARMACOLOGICAL RELEVANCE: Moringa oleifera Lam. (M. oleifera) is a perennial deciduous tree with considerable agricultural and pharmacological value. Nearly all parts of the tree are edible, and nearly all parts are used in traditional medicine. Leaves of M. oleifera have the functions of hypoglycemic (antidiabetic), anti-cancer and anti-oxidant stress, but less research pay attention to the anti-inflammatory effect of M. oleifera leaves. AIM OF THE STUDY: Inflammatory bowel disease (IBD) is a chronic and relapsing inflammatory disorder of the gut with no ideal medication. Here, we investigated the anti-inflammatory effects of aqueous extract of M. oleifera leaves. MATERIALS AND METHODS: Intestinal organoids and mice as in vitro and in vivo models to investigate the effects of aqueous extract of M. oleifera leaves on inflammation induced by TNF-α and dextran sulfate sodium (DSS) respectively. The expression of inflammatory cytokines and proliferation-related genes were evaluated by RT-qPCR, respectively. The compounds in the leaf extract were determined by LC/MS, and network pharmacology approach was employed to predict 54 anti-IBD potential targets of quercetin-3-galactoside (QG) and isoquercitrin (IS). RESULTS: We found that the extract protected against damage to intestinal organoids caused by tumor necrosis factor (TNF-α), and significantly down-regulated the expression of inflammatory cytokines. The extract also suppressed the TNF-α-induced expression of Pcna, c-Myc, and c-Jun. Additionally, oral administration of the extract also ameliorated DSS-induced colon damage (colonic shortening, loss of goblet cells and overall abnormal cellularity), and inhibited the expression of inflammatory cytokines and proliferation-related genes in colitis. By LC/MS we identified nearly 2000 of the compounds in the leaf extract, of the flavonoids identified, QG and IS made up the largest percentage; both have been shown to have anti-inflammatory properties. Moreover, network pharmacology approach was employed to predict 54 anti-IBD potential targets of QG and IS. Gene Ontology (GO) enrichment and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis indicated that the overlapping targets participated in response to oxidative stress and PI3K-Akt signaling pathway respectively. CONCLUSIONS: The present study demonstrated the anti-inflammatory capability, in vitro and in vivo, of the aqueous extract of M. oleifera leaves and suggests its potential phytotherapeutic treatment for IBD.

Moringa oleifera Lam. Leaf improves constipation of rats induced by low-fiber-diet: A proteomics study.

ETHNOPHARMACOLOGICAL RELEVANCE: The leaf of Moringa oleifera Lam., a medicinal and edible herb for thousands of years in Ayurveda, is used as Pancha (na) karma (purgative) during and after the body cleansing process, to treat constipation, reduce cholesterol and body weight. AIM OF STUDY: The aim was to investigate the diarrhea effects and possible mechanism of M. oleifera leaves in constipation rats. MATERIALS AND METHODS: The hot water extract of M. oleifera leaves (WEMOL) was prepared and analyzed using LC-20AT HPLC system. The constipated rat model was induced by feeding with low fiber diet for 21 days. After oral administration of WEMOL for 7 days, the excretion parameter analysis, gastro-intestinal propulsion, histological analysis by HE and Alcian blue staining, and gastrointestinal hormone in rat's digestive tract through ELISA were used to evaluate the laxative effect of WEMOL. Label-free quantitation (LFQ) with LC-MS/MS, bioinformatics and Western blot were used to discover and verify the signal pathways and key proteins of WEMOL related to diarrhea. RESULTS: The contents of isoquercitrin and astragalin were 2.7 mg/g and 1.7 mg/g, respectively in WEMOL. The stool number, weight, and water content of constipation rats were significantly reduced, indicating model had been established successfully. WEMOL (1.25 and 2.5 g/kg) increased water content of feces and the levels of Gas, MTL, NPY in gastric antrum and VIP, SP in colon of constipation rats, improved the muscle layer thickness and mucin secretion of colon. The proteomics revealed a total of 1731 differential proteins and 9 signaling pathways, WEMOL increased the expression of Vamp2, Gnai3, and Prkacb. CONCLUSIONS: The laxative mechanism of WEMOL maybe modulate the signaling pathways mediated by 5-HT and Ach receptors, related to gastrointestinal motility and intestinal fluid secretion. It can be considered as the scientific connotation of Pancha (na) karma of M. oleifera leaves in ayurveda.

Metabolite profiling and biochemical investigation of the antidiabetic potential of Loranthus pulverulentus Wall n-butanol fraction in diabetic animal models.

ETHNOPHARMACOLOGICAL RELEVANCE: Globally, 537 million individuals are estimated to have diabetes. The traditional use of herbs for ameliorating diabetes symptoms is a common practice in Pakistan and use of Loranthus pulverulentus Wall (L. pulverulentus) by local people in Azad Jammu and Kashmir has been reported. AIM OF THE STUDY: In the present study, the antidiabetic potential of standardized n-butanol fraction of leaves of L. pulverulentus Wall, which is a parasite of Dalbergia sisso Roxb was assessed in both alloxan (ALX) and streptozotocin (STZ) diabetic animal models. MATERIALS AND METHODS: Chemical characterization of BF was performed using HPLC, GCMS and UHPLC-MS. The effect of the fraction (250 mg/kg) on insulin, plasma free fatty acids, L-lactate, pyruvate, MDA, HbA1c and glycogen levels in ALX and STZ animal models was determined. Liver and renal profiles were analyzed in the STZ model. Toxicological studies were performed by hemolytic, Ames mutagenicity, DNA protection, and thrombolytic assays. Histopathological analysis of the pancreas, liver, and kidney was performed. RESULTS: BF demonstrated highly significant (p < 0.001) antidiabetic potential in both diabetic models. BF significantly (p < 0.05) improved OGTT results in alloxanized diabetic mice and blocked the absorption of glucose from the gut. A significant (p < 0.001) increase in insulin levels and glycogen content in liver tissue and a decrease in plasma FFA, MDA, HbA1c, L-lactate, and pyruvate levels in STZ-diabetic mice were recorded. GC-MS and chromatographic analysis showed the presence of catechin, eugenol, longifolene, caryophyllene, Ar-tumerone and Geranyl-alpha-terpinene. Various metabolites with antidiabetic potential, including 4-hydroxycinnamyl alcohol 4-D-glucoside, zingerone glucoside, trans-trismethoxy resveratrol-d4, epigallocatechin 3-O-cinnamate, and ß-glucogallin, were identified using UHPLC-MS. Animals treated with BF showed marked improvements in cellular structures of the pancreas, liver and kidneys. This fraction is non-mutagenic and protects the DNA. CONCLUSION: The experimental fraction contained potential antidiabetic bioactive compounds responsible for alleviating diabetes-associated biochemical dysregulation. The fraction increased insulin levels and enhanced glycogen storage in muscles and the liver. It blocked glucose absorption from the intestine and substantially decreased HbA1c, lactate, pyruvate, free fatty acids, lipid, liver and kidney damage. Therefore, the use of BF for the treatment of type-2 diabetes may be beneficial.

Evaluation of Saxifraga stolonifera phenolic extracts as a potential antivenom against Deinagkistrodon acutus venom: In vitro and in vivo studies.

ETHNOPHARMACOLOGICAL RELEVANCE: In the snake-infested mountainous regions of China, Saxifraga stolonifera [L.] Meeb is widely utilized as an immediate remedy for venomous snake bites. However, the scientific understanding of S. stolonifera's efficacy in snakebite treatment remains limited and requires further investigation. AIM OF THE STUDY: The aim of this study was to assess the inhibitory effects of Saxifraga stolonifera phenolic extracts (SSPE) on Deinagkistrodon acutus venom (DAV) and explore the potential of S. stolonifera as a valuable candidate for antivenom development. MATERIALS AND METHODS: We employed our previously optimized extraction conditions to obtain SSPE. In vitro experiments utilizing diverse models were conducted to assess the inhibitory effects of the extracted phenolic compounds on DAV, specifically targeting phospholipase A2 (PLA2), proteolytic, fibrinolytic, and hyaluronidase enzymes. Furthermore, in vivo investigations were conducted to evaluate the inhibitory potential of the extracted compounds against DAV-induced hemorrhagic and edematogenic activity. To elucidate the chemical composition of the phenolic extracts, Ultra Performance Liquid Chromatography-mass spectrometry (UPLC-MS) analysis was performed. RESULTS: Our in vitro inhibition study showed that S. stolonifera was able to inhibit the activities of PLA2 enzyme, proteolytic enzyme, hyaluronidase and fibrinogenolytic. The median effective dose (ED50) values of SSPE for inhibiting PLA2 enzyme, proteolytic enzyme and hyaluronidase activities were 0.115 mg/mL, 0.026 mg/mL and 0.238 mg/mL, respectively. The DAV-induced hemorrhagic and edematogenic effects of the SSPE were also successfully inhibited in vivo, and the high SSPE concentration was able to completely inhibit the hemorrhage and edema. It is noteworthy that the mice suffered no harm from the high SSPE concentration. The composition analysis showed that the phenolic substances contained in SSPE are gallic acid, protocatechuic acid, chlorogenic acid, rutin, kaempferol-3-O-ɑ-L-rhamnoside, kaempferol-3-O-ß-D-glucopyranoside, quercetin and kaempferol. CONCLUSIONS: This study provides scientific validation of the inhibitory efficacy of S. stolonifera as an emergency treatment for venomous snake bites, offering a theoretical foundation for future drug development strategies targeting snakebite.

Skin microbiota metabolism of natural products from comfrey root (Symphytum officinale L.).

ETHNOPHARMACOLOGICAL RELEVANCE: Comfrey root (Symphytum officinale L., Boraginaceae) has been used in folk medicine for a long time to treat different diseases. It is recommended for swellings, phlebitis, contusions, gastro-duodenal ulcers, respiratory diseases, and metrorrhagia. Currently, preparations from S. officinale are only topically used due to its wound-healing effects, and for reducing inflammation and the treatment of broken bones, tendon damage, painful joints and muscles. Although it is a widespread plant material, little is known about the interaction of externally applied preparations of comfrey with the human skin microbiome. AIM OF THE STUDY: The study aims to determine the interaction between human skin microbiota and the comfrey root extracts, by monitoring the biotransformation of the constituents present in the extract and evaluating changes in the population of the skin microbiota in an ex vivo setting. MATERIAL AND METHODS: The comfrey root extract was incubated with the human skin microbiota from ten healthy donors. The UHPLC-DAD-MSn analysis determined the composition of the raw extract and the microbial metabolites. Bacterial genomic DNA was extracted and examined by amplification sequencing of the 16S rDNA to determine changes in the bacterial composition. RESULTS: The hydroethanolic extract of comfrey root primarily consists of phenolic acids, pyrrolizidine alkaloids, and their derivatives, and lignans. The natural products present in the extract underwent biodegradation by the skin microbiota, leading to the formation of smaller molecules. It was observed that the skin microbial metabolism primarily focused on modifying the derivatives of pyrrolizidine alkaloids. It resulted in the production of deacetylated and deesterificated compounds. However, it did not lead to the conversion of these compounds into free alkaloids. CONCLUSIONS: The microbiota-triggered biotransformation of the comfrey root extract was observed. A few N-oxides were metabolized to deacetylated and deesterificated forms in ex vivo conditions. It suggests that the intermittent external applications of comfrey preparations perchance are unlikely to pose a substantial risk. While it even may serve as a potential factor influencing the extract activity in treating skin diseases.

Effect of chin brick tea [Camellia sinensis (L.) Kuntze] on lipid metabolism and inflammation by modulating intestinal flora and bile acids in mice with non-alcoholic fatty liver disease.

ETHNOPHARMACOLOGICAL RELEVANCE: Tea (Camellia sinensis) has been consumed for centuries as a traditional remedy for various metabolic diseases. The pharmacological mechanisms of many conventional medicines, including tea, often need to be clarified. Chin brick tea is a unique Chinese black tea grown in Hubei, China, rich in tea elements such as tea polyphenols and tea polysaccharides. AIM OF THE STUDY: We focus on the effects of commercial chin brick tea on non-alcoholic fatty liver disease by altering intestinal flora and its metabolite, bile acids. MATERIALS AND METHODS: Targeted UPLC-MS/MS was employed to quantify the tea elements in commercial chin brick tea. In this study, we performed an integrated approach of animal experiments, 16 S rDNA, and ultra-performance liquid chromatography-tandem mass spectrometry to explore the potential mechanism of action of chin brick tea in preventing non-alcoholic fatty liver disease (NAFLD). RESULTS: After 14 weeks of administration, CBT extract could signiffcantly decrease the levels of body weight, liver weight, LDL-C, TC, ALT, IL-1ß and IL-18, and slight increase HDL-C levels in NAFLD mice. The results indicated that the interventional impact of CBT with high-fat diet-induced NAFLD might depend on intestinal flora and its metabolites bile acids. Moreover, sequencing of 16 S rRNA genes demonstrated that CBT could signiffcantly improve the intestinal flora disorder of NAFLD mice. Speciffcally, CBT increased the levels of Lactobacillus, Alloprevotella, and Ruminococcaceae, while reducing the levels of Bacteroides in NAFLD mice. Then, a total of 23 bile acids were identified, 17 differential bile acids were obtained by screening, and CBT increase the primary bile acids/secondary bile acids ratio in NAFLD mice. Additionally, correlation analysis revealed that Bacteroides was negatively correlated with DCA and ωMCA, Lactobacillus was positively correlated with DCA and ωMCA, Bacteroides was positively correlated with NAFLD, Lactobacillus was negatively associated with NAFLD, and DCA and ωMCA were negatively correlated with NAFLD. CONCLUSION: CBT extract has a good interventional impact on NAFLD mice. The mechanism by which this extract exerts its action is, at least partly, related to its regulation of intestinal flora and its metabolites bile acids.

Antiulcer activity and mechanism of action of the hydroethanolic extract of leaves of Terminalia argentea Mart. In different in vivo and in vitro experimental models.

ETHNOPHARMACOLOGICAL RELEVANCE: Terminalia argentea Mart. (Combretaceae) is a deciduous tree commonly found in Brazil, Bolivia, and Paraguay. It occurs in all regions of Brazil and is widespread in the Amazon, Cerrado, Pantanal, Atlantic Rain Forest, and Caatinga Biomes. In the traditional medicine of Brazil, people widely use tea or decoction of its leaf materials for treating gastritis, ulcers, wound healing, and inflammation. AIM OF THE STUDY: The current study aims to evaluate the gastroprotective and ulcer-healing activities of the hydroethanolic extract of T. argentea leaves (HETa) and investigate the underlying mechanisms of action through in vivo and in vitro experiments. METHODS: We extracted the leaves of T. argentea with a 70% hydroethanolic solution (HETa) and performed phytochemical analysis using high-performance liquid chromatography (HPLC) and electrospray ionization mass spectrometry (ESI-MSn). We researched the antiulcer activity using in vivo and in vitro experiments, administering three doses (2, 10, and 50 mg/kg) and different concentrations of 1, 5, and 20 µg/mL, respectively. We verified the acute antiulcer activity using chemical models (acidified ethanol (EtOH/HCl) and indomethacin (IND)) and physiological models (water-immersion stress (WRS)). To induce chronic ulcers, used acetic acid and treated the animals for seven days. To investigate the mechanism of action, conducted assays of antioxidant activity, measured the dosage of inflammatory cytokines, quantified mucus, treated with inhibitors (IND, L-NAME, glibenclamide, and yohimbine), performed histopathological analysis, and measured gastric acid secretion. Furthermore, we performed in vitro experiments on murine macrophage cell lines (RAW 264-7 cells) to quantify nitrite/nitrate and cytokine production and on V79-4 cells to verify cell proliferation/migration. RESULTS: We conducted HPLC and ESI-MSn analyses to obtain a fingerprint of the chemical composition of the HETa, revealing the presence of phenolics (caffeoyl ellagic acid), flavonoids (rutin, quercetin xyloside, quercetin rhamnoside, quercetin glucoside, quercetin galloyl xyloside, quercetin), and tannins (terminalin), respectively. The three doses of HETa reduced acute and chronic ulcers in different models. The mechanism of action involves increasing mucus production and angiogenesis, and it partially involves prostaglandins, nitric oxide, K+ATP channels, and α2-adrenergic receptors. HETa also exhibited antioxidant potential, reducing myeloperoxidase (MPO) activity, and increasing glutathione (GSH) levels. Moreover, it demonstrated anti-inflammatory action by reducing nitrite/nitrate levels and pro-inflammatory cytokine concentrations in vivo, and it increased in vitro proliferation/migration of fibroblasts. CONCLUSIONS: The study shows that HETa presents a potent preventive and curative antiulcer effect in different ulcer models, supporting the popular use of homemade preparations of T. argentea leaves. The preventive and gastric healing ulcer activity of HETa involves multiple targets, including increasing the gastric mucus barrier, antioxidant defenses, and anti-inflammatory effects on gastric mucosa repair. Phytochemical analysis identified the presence of phenolic compounds, flavonoids, and tannins in HETa, and the antiulcer activity may be attributable to the combined effect of these constituents.

Pharmacological disadvantages in the spasmolytic effects by using the mixture known as "three toronjiles" in folk medicine.

ETHNOPHARMACOLOGY RELEVANCE: In Mexico, Agastache mexicana subsp. mexicana (PT) and subsp. xolocotziana (WT), and Dracocephalum moldavica (BT), are used together as the "three toronjiles" to treat gastrointestinal spasms. AIM OF STUDY: To evaluate if the spasmolytic activity of these three medicinal species is better in combination or in an individual manner. MATERIALS AND METHODS: Spasmolytic effect of PT, WT and BT alone or combinate were evaluated in rings of the guinea-pig ileum contracted with potassium chloride (KCl), electrical field stimulation (EFS), or acetylcholine (ACh). Chemical analysis by thin layer and ultra-high pressure liquid chromatography of the aqueous extracts of each species were done for their comparison, and their acute toxicity were determined in mice. RESULTS: PT and WT diminished in a dose-dependent manner the contractions induced by KCl, EFS, and ACh. Whereas BT did not altered contractions in any experimental protocol. A combination of the PT (EC50) and WT (EC50) diminished the contractions induced by KCl or EFS. Interestingly, the addition of BT extract (10 µg/ml) to the combination (PT EC30 + WT EC30) blocked the inhibitory effect produced on the contracted tissue in the presence of KCl, EFS, or ACh. Moreover, addition of BT extract (100 µg/ml) to the same combination blocked the inhibitory effect on the pre-contracted tissue only in the presence of EFS. None of the aqueous extracts produced toxicity in its individual administration in mice. Chemical analysis demonstrated similarities between PT and WT, but differences with BT. CONCLUSIONS: The results of this study confirmed that either combined or by themselves aqueous extracts of PT and WT produced a spasmolytic action on guinea pig ileum, suggesting that this combination of medicinal plants could relieve gastrointestinal diseases in human, but when BT aqueous extract is added to those obtained with PT and WT, the spasmolytic activity diminished or even was blocked. Our results give evidence that mixture of several plants might produce disadvantages in the medicinal properties of their individual activity.

Withania somnifera ameliorates sexual arousal and impotence in stressed sexually sluggish male rats by modulating neurotransmitters and NO/cGMP/PDE5α pathway.

ETHNOPHARMACOLOGICAL RELEVANCE: Psychological stress is a growing global threat to male sexual potency and erection efficiency. Withania somnifera (L.) Dunal (WS), also known as Ashwagandha, is a well-known Ayurvedic herb. The roots of Withania somnifera improve the body's ability to handle stress, strengthen the immune system, promote healthy ageing, and have aphrodisiac properties with male sexual stimulation effects. Despite its widespread acceptance as an Ayurvedic stress-relieving drug with beneficial effects on male reproductive health, Withania somnifera has yet to be studied for its potential role in improving the sexual arousal and erectile dysfunction of psychologically stressed sexually sluggish males. AIM OF THE STUDY: To investigate the therapeutic effects of purified root powder of Withania somnifera on sexual behaviour and erectile efficiency in stressed sexually sluggish male rats. MATERIALS AND METHODS: Sexually sluggish male rats were screened by premating tests after being exposed to a psychological stressor, restraint stress, 3 h/day for 30 days. Subsequently, these rats were treated with purified root powder of WS (150 or 300 mg/kg/day-PO) or sildenafil (5 mg/kg/day-PO) for 30 days. The rats were sacrificed after 24 h of the last treatment, and the effects on various factors related to sexual behaviour, penile histomorphology, serum hormones, and neurotransmitters associated with sexual arousal and penile erection were examined. RESULTS: WS treatment improves prosexual and sexual behaviour in psychologically stressed sexually sluggish male rats by increasing non-contact erections and mounts, intromission, and ejaculation frequencies, while decreasing sexual exhaustion by decreasing post-ejaculation intervals and latencies. WS also modulates neurotransmitters and hormones associated with sexual desire and stress, including dopamine, serotonin, corticosterone, and prolactin. Additionally, there was also a dose-dependent increase in serum LH, FSH, and testosterone levels. The administration of WS to sexually sluggish rats resulted in significant improvements in penile histomorphology, specifically by increasing the ratio of smooth muscle (SM) to collagen. Furthermore, in sexually sluggish rats, WS treatment increased the expression of markers associated with penile erection facilitation, such as nNOS, eNOS, p-Akt, nitric oxide, acetylcholine, and cGMP. Notably, WS treatment decreased the expression of penile PDE5α in these rats in a dose-dependent manner. Remarkably, the therapeutic effects of WS are comparable to those of sildenafil. CONCLUSIONS: Purified root powder of Withania somnifera was found to improve sexual arousal and erection efficiency in stressed, sexually sluggish male rats. This improvement was achieved by modulating the HPG and HPA axes as well as the NO/cGMP/PDE5α pathway involved in penile erection. Thus, our findings strongly support the potent therapeutic potential of purified root powder of WS in improving the sexual health of stressed sexually sluggish rats.

Rosemary (Rosmarinus officinalis L.) hydrosol based on serotonergic synapse for insomnia.

ETHNOPHARMACOLOGICAL RELEVANCE: Rosemary (Rosmarinus officinalis L.) has been widely used as a traditional remedy for insomnia, depression and anxiety in China and Western countries. Modern pharmacological studies have shown that rosemary has important applications in neurological disorders. However, the mechanism of action of rosemary hydrosol in the treatment of insomnia is not known. AIMS OF THE STUDY: Insomnia is closely linked to anxiety and depression, and its pathogenesis is related to biology, psychology, and sociology. Rosemary is a natural plant that has been used to treat insomnia and depression and has good biological activity, but its material basis and mechanism for the treatment of insomnia are not clear. Here, we report on the role of aqueous extracts of rosemary in the treatment of insomnia. MATERIALS AND METHODS: The study was based on network pharmacology, using a combination of RNA-sequencing, "quantity-effect" weighting coefficients, and pharmacodynamic experiments. DL-4-chlorophenylalanine (PCPA) was intraperitoneally injected into SD rats to replicate the insomnia model with a blank, model, diazepam, and rosemary hydrosol low-, medium-, and high-dose groups were set up for the experiment. The key pathways in the treatment of insomnia with rosemary hydrosol were analyzed by molecular docking, open field assay, ELISA, western-Blot, Rt-PCR, and immunohistochemical assay. RESULTS: Rosemary hydrosol was analyzed by GC-MS to identify 19 components. 1579 differential genes were obtained by RNA-Seq analysis, 533 targets for rosemary hydrosol and 2705 targets for insomnia, and 29 key targets were obtained by intersection. The KEGG results were ranked by "quantity-effect" weighting coefficients, resulting in serotonergic synapse was the key pathway for the treatment of insomnia with rosemary hydrosol. Molecular docking results showed that 1,7,7-trimethylbicyclo[2.2.1] heptan-2-one, 3-methyl-4-isopropylphenol, caryophyllene, and citronellol of rosemary hydrosol acted synergistically to achieve a therapeutic effect on insomnia. Caryophyllene acts on the HTR1A target by upregulating 5-HT1AR, leading to increased 5-HT release, and upregulation of ADCY5, cAMP, PKA and GABAA at serotonergic synapses; citronellol upregulated ADCY5 and 1,7,7-trimethylbicyclo[2.2.1] heptan-2-one, and 3-methyl-4-isopropylphenol up-regulated GABAA to improve insomnia symptoms. In open-field experiments, ELISA kits (5-HT, GABA, and DA), Western-blotting, Rt-PCR and immunohistochemical assay experiments, insomnia rats in the low-, medium- and high-dose groups of rosemary hydrosol showed different degrees of improvement compared with the model group. CONCLUSIONS: It was shown that rosemary hydrosol may exert its therapeutic effects on insomnia through serotonergic synapses by combining RNA-Seq, "quantity-effect" weighting coefficients network pharmacology and pharmacodynamic experiments. We have provided a preliminary theoretical study for the development of rosemary hydrosol additive into a beverage for the treatment of insomnia, but it needs to be studied in depth. This study was conducted in rats and the results have limitations and may not apply to humans.

Buddleja officinalis Maxim.: A review of its botany, ethnopharmacology, phytochemistry, pharmacology, and therapeutic potential for ophthalmic diseases.

ETHNOPHARMACOLOGICAL RELEVANCE: Buddleja officinalis Maxim. (B. officinalis), commonly known as "Menghua" "Yangerduo" is a widely recognized traditional herbal medicine in China, Korea, and Vietnam. For thousands of years, it has been used to treat dry eye disease, conjunctivitis, keratitis, eye ulcers, eye pain, cough, asthma, hemoptysis, and other medical conditions. AIM OF THE REVIEW: This review article aims to provide a concise summary of the botany, ethnopharmacology, phytochemistry, pharmacology, medicinal potential, and application of B. officinalis in treating ophthalmic diseases and critically evaluates the existing literature to establish a scientific basis for its reasonable utilization and further investigation. MATERIALS AND METHODS: The information reviewed in this study was collected from various electronic resources, including the Web of Science, PubMed, and Google Scholar. RESULTS: To date, 80 structurally diverse compounds have been isolated and characterized from B. officinalis, primarily flavonoids, phenylethanoids, triterpenoids, and monoterpenes. Extracts and compounds derived from B. officinalis have been reported to possess broad pharmacological effects including anti-dry eye disease, anti-inflammation, anti-oxidation, anti-diabetes, anti-obesity, improving osteoporosis and treatment of skin diseases. This review provides a reference for the future studies on of B. officinalis. CONCLUSIONS: As a natural medicinal plant, B. officinalis is worthy of further development in botany, ethnopharmacology, phytochemistry, pharmacology, and therapeutic potential for ophthalmic diseases. Although some components have demonstrated multiple pharmacological activities, their mechanisms of action remain unclear. Further studies on the underlying molecular basis and mechanism of action are warranted.

A blend of Withania somnifera (L.) Dunal root and Abelmoschus esculentus (L.) Moench fruit extracts relieves constipation and improves bowel function: A proof-of-concept clinical investigation.

ETHNOPHARMACOLOGICAL RELEVANCE: Withania somnifera (L.) Dunal (WS) and Abelmoschus esculentus (L.) Moench (AE) are known as Ashwagandha and Okra, respectively, important herbs in traditional medicine for their diverse therapeutic values. WS root is an adaptogen that relieves stress and anxiety and promotes sleep. AE fruit or Okra is widely consumed as a vegetable and is traditionally used to treat diabetes, gastric irritations, ulcers, and obesity. AIM OF THE STUDY: The present randomized, double-blind, placebo-controlled study aimed to establish a proof-of-concept evaluating the efficacy and tolerability of a proprietary blend of standardized extracts of WS root and AE fruit, CL18100F4 in relieving constipation and improving quality of life in adults. MATERIALS AND METHODS: Forty-eight male and female participants (age: 25-60 years) with functional constipation (following Rome-III criteria) were randomized into placebo, 300 or 500 mg of CL18100F4 groups, and supplemented for fourteen consecutive days. RESULTS: CL18100F4 supplementation significantly (p < 0.0001) reduced the Patient Assessment of Constipation-Symptoms (PAC-SYM), Patient Assessment of Constipation-Quality of Life (PAC-QOL), and Gastrointestinal Symptom Rating Scale (GSRS) scores. CL18100F4 supplementation improved sleep quality and reduced stress (p < 0.0001). At the end of the study, CL18100F4-500 subjects showed significant increases in serum serotonin, gastrin, and interleukin-10 and decrease in interleukin-6 and cortisol levels. Participants' hematology, total blood chemistry, vital signs, and urinalysis parameters were within the normal ranges. No adverse events were reported. CONCLUSIONS: This short-duration, single-site clinical investigation demonstrates that CL18100F4 supplementation is tolerable, helps relieve constipation, reduces stress, and improves gastrointestinal function, sleep quality, and general wellness in adults. TRIAL REGISTRATION: Clinical Trials Registry- India (CTRI/2020/11/029320); Registered on 24/11/2020. Available at: http://ctri.nic.in/Clinicaltrials/showallp.php?mid1=49391&EncHid=&userName=CL18100F4.

Antioxidant, antitumoral, antimetastatic effect and inhibition of collagenase enzyme activity of Eleutherine bulbosa (Dayak onion) extract: In vitro, in vivo and in silico approaches.

ETHNOPHARMACOLOGICAL RELEVANCE: Eleutherine bulbosa (Mill.) Urb., known in Brazil as "marupazinho", is a medicinal plant native to the Amazon region. The bulbs of this species are traditionally used in the form of tea or consumed in natura (salads) for the treatment of hypertension, diabetes, gastrointestinal problems, breast cancer, and female fertility. It has been reported that this species possess cytotoxic compounds with anticancer action and limited underlying mechanisms. AIM OF THE STUDY: This study aimed to analyze extract of E. bulbosa bulbs and evaluate antioxidant activity, antitumor and antimetastatic effects against murine B16F10-Nex2 melanoma cells, and collagenase inhibitory activity by in vitro, in vivo, and in silico approaches. MATERIALS AND METHODS: Determination of total polyphenols, flavonoids and anthocyanins content were performed. In addition, high performance liquid chromatography-mass spectrometry (HPLC-MS) was carried out to identify phytoconstituents from extract. Antioxidant evaluation was performed using DPPH radical scavenging, ferric ion reducing antioxidant power (FRAP), Thiobarbituric acid reactive species (TBARS) and nitric oxide (NO) tests. Antitumoral and antimetastatic activities of extract on murine B16F10-Nex2 melanoma cells were determined and inhibitory activity on collagenase was evaluated. Molecular interactions between compounds and DNA or collagenase was evaluated by molecular docking analyses. RESULTS: Phytochemical evaluation demonstrated the presence of polyphenols, flavonoids and anthocyanins, and HPLC-MS identified the major presence of eleutherin, isoeleutherin and eleutherinol. Antioxidant evaluation showed that the extract present significant activity in all methods evaluated. In silico assay demonstrated interaction between bioactive compounds and DNA or collagenase. In addition, extract exhibited antitumor and antimetastatic actions promoted by melanoma cells and showed collagenase inhibitory activity. CONCLUSIONS: The results showed that E. bulbosa bulb extract contains bioactive compounds as flavonoids, anthocyanins and quinones of which may be responsible for the antioxidant, antitumor, antimetastatic and collagenase enzyme inhibitory activity observed in this study by in vivo, in vitro and in silico bioassays.

Himalayan Pyracantha crenulata (D.Don) M.Roem. leaf and fruit extracts alleviate algesia through COX-2 and Mu-opioid receptor mediated pathways.

ETHNOPHARMACOLOGICAL RELEVANCE: Pyracantha crenulata (D.Don) M.Roem., a plant of high nutritional and medicinal value is traditionally employed for its analgesic property in joint and body pain in the Kumaun region of Western Himalaya. AIM OF THE STUDY: To validate the traditional claims for analgesic property of Pyracantha crenulata. METHODS: Hydroethanolic extract of P. crenulata leaves and fruits were tested for their analgesic potential in rodent models for algesia by tail immersion test, tail flick test, Eddy's hot plate model, and formalin induced paw irritation test in Wistar rats. Molecular docking and dynamics studies were also performed to understand the possible mechanisms. RESULTS: Both P. crenulata fruit extract and leaf extract exhibited significant amelioration in all the tested experimental models of algesia acting through central and peripheral mechanisms. The efficacy in reducing nociception was found comparable to diclofenac that was used as a reference standard. Molecular docking and dynamic simulation studies further established binding affinity of gallic acid (confirmed to be present in P. crenulata leaf extract through HPTLC profiling) with cyclooxygenase-2 (COX-2) and mu-opioid receptors, suggesting the modulatory effect of these extracts on COX-2 and mu-opioid receptors in combating algesia. CONCLUSION: P. crenulata extracts produce analgesic effects plausibly through COX-2 and mu-opioid receptor mediated pathways.