Ethnopharmacological Evidence Research Articles

Deciphering the multi-functional role of Indian propolis for the management of Alzheimer's disease by integrating LC-MS/MS, network pharmacology, molecular docking, and in-vitro studies.

The conventional one-drug-one-disease theory has lost its sheen in multigenic diseases such as Alzheimer's disease (AD). Propolis, a honeybee-derived product has ethnopharmacological evidence of antioxidant, anti-inflammatory, antimicrobial and neuroprotective properties. However, the chemical composition is complex and highly variable geographically. So, to leverage the potential of propolis as an effective treatment modality, it is essential to understand the role of each phytochemical in the AD pathophysiology. Therefore, the present study was aimed at investigating the anti-Alzheimer effect of bioactive in Indian propolis (IP) by combining LC-MS/MS fingerprinting, with network-based analysis and experimental validation. First, phytoconstituents in IP extract were identified using an in-house LC-MS/MS method. The drug likeness and toxicity were assessed, followed by identification of AD targets. The constituent-target-gene network was then constructed along with protein-protein interactions, gene pathway, ontology, and enrichment analysis. LC-MS/MS analysis identified 16 known metabolites with druggable properties except for luteolin-5-methyl ether. The network pharmacology-based analysis revealed that the hit propolis constituents were majorly flavonoids, whereas the main AD-associated targets were MAOB, ESR1, BACE1, AChE, CDK5, GSK3β, and PTGS2. A total of 18 gene pathways were identified to be associated, with the pathways related to AD among the topmost enriched. Molecular docking analysis against top AD targets resulted in suitable binding interactions at the active site of target proteins. Further, the protective role of IP in AD was confirmed with cell-line studies on PC-12, in situ AChE inhibition, and antioxidant assays.

A Review of Healing Potential of Indian Flora: An Ethnopharmacological Evidence of Wound Healing Activity

Introduction: Healing a wound involves a complex series of coordinated chemical and cellular events to restore the integrity of skin and underlying tissues. Many people prefer using medicinal plants as an alternative therapy for this purpose globally. Objective: The focal point of the review is based on the ethnopharmacological profiling and validation of plants. The reviewed plants belong to the India region and have promising wound healing activity which has been scientifically proven by in vivo as well as in vitro studies. Numerous phyto extracts and their secondary metabolites are known as promising alternatives for wound healing agents due to the existence of miscellaneous dynamic constituents, ease of access, and their inadequate side effects. Methods: The plant profile was explored from various sources including scientific search engines such as Google Scholar, Niscare repository, Scopus, Pubmed, Sciencedirect, Sci-Finder, and Springer including reports and thesis from Proquest. Keywords such as wound healing property, plant extract, and animal models were employed in combination with the ethnopharmacological claim, pharmacological activity, etc. Conclusion: This review provides comprehensive data from published research work on medicinal plants over 10 years, focusing on wound healing activity and its ethnopharmacological profiling from different tribal belts. This review features the potential of medicinal plants to contribute to drug discovery as a compelling wound healer which may help in the discovery of a new drug molecule with a maximum margin of safety. However further studies are needed such as novel formulation, and target-based drug delivery systems by performing clinical studies for the scientific exploration and validation from laboratory to industry for societal value.

Clinical pharmacokinetic evaluation of Withania somnifera (L.) Dunal root extract in healthy human volunteers: A non-randomized, single dose study utilizing UHPLC-MS/MS analysis

Ethnopharmacological relevanceWithania somnifera (L.) Dunal; (Solanaceae), commonly known as Ashwagandha, is one of the most significant medicinal herbs in ‘Ayurveda’, a traditional Indian medicine used for centuries with evidence in scriptures. Ashwagandha was mentioned in old Ayurvedic medical literature such as Charaka Samhita and Sushruta Samhita for improving weight and strength, with multiple citations for internal and exterior usage in emaciation and nourishing the body. Ethnopharmacological evidence revealed that it was used to relieve inflammation, reduce abdominal swelling, as a mild purgative, and treat swollen glands. The root was regarded as a tonic, aphrodisiac, and emmenagogue in the Unani tradition of the Indian medicinal system. Further, Ashwagandha has been also described as an Ayurvedic medicinal plant in the Ayurvedic Pharmacopoeia of India extending informed therapeutic usage and formulations. Despite the widespread ethnopharmacological usage of Ashwagandha, clinical pharmacokinetic parameters are lacking in the literature; hence, the findings of this study will be relevant for calculating doses for future clinical evaluations of Ashwagandha root extract. AimThis study aimed to develop a validated and highly sensitive bioanalytical method for quantifying withanosides and withanolides of the Ashwagandha root extract in human plasma to explore its bioaccessibility. Further to apply a developed method to perform pharmacokinetics of standardized Withania somnifera (L.) Dunal root extract (WSE; AgeVel®/Witholytin®) capsules in healthy human volunteers. MethodsA sensitive, reliable, and specific ultra-high pressure liquid chromatography-mass spectrometry (UHPLC-MS/MS) method was developed and validated for the simultaneous quantification of five major withanosides and withanolides (withanoside IV, withanoside V, withanolide A, withaferin A, and 12-deoxy-withastramonolide) in human plasma. Further for the study, eighteen healthy male volunteers (18–45 years) were enrolled in a non-randomized, open-label, single period, single treatment, clinical pharmacokinetic study and given a single dose (500 mg) of WSE (AgeVel®/Witholytin®) capsules containing not less than 7.5 mg of total withanolides under fasting condition. Later, pharmacokinetic profiles were assessed using the plasma concentration of each bioactive constituent Vs. time data. ResultsFor all five constituents, the bioanalytical method demonstrated high selectivity, specificity, and linearity. There was no carryover, and no matrix effect was observed. Furthermore, the inter-day and intra-day precision and accuracy results fulfilled the acceptance criteria. Upon oral administration of WSE capsules, Cmax was found to be 0.639 ± 0.211, 2.926 ± 1.317, 2.833 ± 0.981, and 5.498 ± 1.986 ng/mL for withanoside IV, withanolide A, withaferin A, and 12-deoxy-withastramonolide with Tmax of 1.639 ± 0.993, 1.361 ± 0.850, 0.903 ± 0.273, and 1.375 ± 0.510 h respectively. Further, withanoside V was also detected in plasma; but its concentration was found below LLOQ. ConclusionThe novel and first-time developed bioanalytical method was successfully applied for the quantification of five bio-active constituents in human volunteers following administration of WSE capsules, indicating that withanosides and withanolides were rapidly absorbed from the stomach, have high oral bioavailability, and an optimum half-life to produce significant pharmacological activity. Further, AgeVel®/Witholytin® was found safe and well tolerated after oral administration, with no adverse reaction observed at a 500 mg dose.

Ethnopharmacology of Botanical Galactagogues and Comprehensive Analysis of Gaps Between Traditional and Scientific Evidence

The functional potential of plants and plant-based components have been used in different cultures since time immemorial to promote milk production in women. Conditions like agalactia or insufficient production of breast milk due to various barriers intended to be a greater risk of weight loss for neonates and prompting for supplementation of infant formula. Multiple plants are traditionally used worldwide as galactagogues during the lactation period. This study aims to extract information on traditionally used galactagogue plants and compare their ethnopharmacological evidence with scientific evidence. This will help to understand the gaps in the ethnopharmacological and scientific data and thus provide future research information.Information on traditional and scientific studies was collected and analyzed on galactagogues. The ethnopharmacological data of the focused plant species were analyzed for part used, formulations, and region of its uses. All cultures traditionally used natural products as galactagogue from times immemorial, and due to scientific advances, these have also been seen as commercial products. However, because of the limited studies, it is of interest to standardize the doses, and composition of bioactive components and study the mechanism of action, its side effects, and interaction with food. This is a forward-looking research area that could be projected for manufacturing herbal formulations for lactating mothers.

A methanolic extract of Eclipta prostrata (L.) L. decreases inflammation in a murine model of chronic allergic asthma via inhibition of the NF-kappa-B pathway

Ethnopharmacological relevanceEclipta prostrata (L.) L. is a medicinal plant used by many ethnic groups in Brazil to treat respiratory diseases, hepatitis and the bites of venomous animals. A methanolic extract of E. prostrata (MEEP), the major components of which are wedelolactone (WED) and demethylwedelolactone (DMW), exhibited anti-inflammatory activity in acute asthma models but the effects on lung inflammation and the mechanisms of action of MEEP in a chronic asthma model are not known. Aim of the studyTo study the effects of MEEP in vivo using a chronic ovalbumin (OVA)-induced allergic asthma model in mice. Materials and methodsThe identities of WED and DMW in MEEP were confirmed and the concentrations determined by liquid chromatography and tandem mass spectrometry. Male Balb/c mice were sensitized and challenged with OVA and experimental animals were treated with MEEP (100, 250 or 500 mg/kg) while control animals were treated with dexamethasone (2 mg/kg) or normal saline. Bronchial hyperresponsiveness, total and differential cell counts in bronchoalveolar lavage (BAL), and the production of Th2 cytokines in lung homogenates were assessed. Lung inflammation and mucus production were evaluated by histological analysis while nuclear factor kappa-B (NF-κB) activation was assessed immunohistochemically. ResultsConcentrations of WED and DMW in MEEP were 5.12% and 1.04%, respectively. Treatments with MEEP (250 or 500 mg/kg) significantly decreased bronchial hyperresponsiveness, reduced total cell and eosinophil counts in BAL and IL-4 concentrations in lung homogenate, and inhibited NF-κB activation. Treatment with MEEP at 500 mg/kg reduced the level of IL-5 in lung homogenates but did not decrease IL-13 concentration or mucus production. ConclusionsMEEP attenuated bronchial hyperresponsiveness and decreased lung and airway inflammation in a chronic asthma model in mice. The mechanism of action involves inhibition of NF-κB activation, most likely associated with the presence of the coumestans WED and DMW. These results support the ethnopharmacological evidence for the use of E. prostrata against asthma and other respiratory inflammatory diseases.

Phytochemical constituents, antioxidant and antibacterial activities of Plectocephalus varians (A. Rich.) C. Jeffrey ex Cufod root extracts

Plants have been used to treat diverse types of diseases in different cultural groups around the globe. In this regard, the root of Plectocephalus varians (P. varians) is claimed to have a beneficiary effect in treating cancer and hemorrhoids in Ethiopia. Hence, this study aimed at the phytochemical investigation, antioxidant, and antibacterial activities of n-hexane, acetone, and methanolic extracts of P. varians root. The different crude extracts of P. varians were obtained through maceration technique. The total phenolic content (TPC) and total flavonoid contents (TFC) of the extracts were estimated using Folin-Ciocalteu Reagent (FCR) and aluminum chloride colorimetric assays, respectively. The antioxidant activities of the extracts were determined by 2,2-diphenyl-1-picrylhydrazyl (DPPH) free radical scavenging and ferric reducing antioxidant power (FRAP) assays. The antibacterial activities of the extracts were assessed by using disc diffusion method. The results echoed the presence of alkaloids, saponins, flavonoids, steroids, phenols, tannins, anthraquinones, terpenoids, polyphenols, and glycosides in the root of P. varians. The methanolic root extract (MRE) had the highest TPC (107.18 mg GAE/g) and TFC (120.194 mg QE/g) followed by acetone root extract (ARE) (TPC = 98.68 mg GAE/g; TFC = 64.038 mg QE/g) and n-hexane root extract (HRE) (TPC = 12.39 mg GAE/g; TFC = 9.917 mg QE/g). The DPPH radical scavenging and FRAP assays demonstrated the antioxidant effects of HRE (IC50 = 681.75 ppm; EC50 = 60.65 ppm), ARE (IC50 = 165.73 ppm; EC50 = 51.67 ppm) and MRE (IC50 = 132.06 ppm; EC50 = 30.97 ppm) of P. varians. Furthermore, the root fractions elicited pronounced dose-dependent growth inhibition against Staphylococcus aureus, Streptococci pyogenes, Escherichia coli, and Klebsiella pneumoniae with mean zone of inhibition (MZI) ranging from 11 ± 0.38 to 20 ± 0.04 mm at 800 ppm. Overall, the present study provides ethnopharmacological evidence suggesting the medicinal importance of P. varians. The results also call for further bioassay-guided phytochemical screening and in vitro and/or in vivo bioactivity testing.

Phenolic profile, in vitro antimicrobial and in vivo diuretic effects of endemic wild thyme Thymus comosus Heuff ex. Griseb. (Lamiaceae) from Romania.

Thymus comosus Heuff ex. Griseb. (Lamiaceae) is a wild thyme species endemic for Romanian Carpathian areas, frequently collected as substitute for collective herbal product Serpylli herba, cited as antibacterial and diuretic remedy in traditional medicine. The present study aimed to evaluate the in vivo diuretic effect and in vitro antimicrobial properties of three herbal preparations (infusion-TCI, tincture-TCT and an hydroethanolic extract prepared through an optimized ultrasound-assisted method-OpTC) obtained from the aerial parts of T. comosus Heuff ex. Griseb, also evaluating their comprehensive phenolic profile. In vivo diuretic effect was tested using Wistar rats treated orally with each herbal preparation (125 and 250mg/kg dispersed in 25ml/kg isotonic saline solution) and quantified based on cumulative urine output (ml), diuretic action and diuretic activity. Additionally, sodium and potassium excretion were monitored using a potentiometric method with selective electrodes. In vitro antibacterial and antifungal activities were assessed using p-iodonitrotetrazolium chloride assay against six bacterial strains and six fungal strains by monitoring minimum inhibitory concentration (MICs), minimum bactericidal concentrations (MBCs) and minimum fungicidal concentrations (MFCs). Finally, phenolic profile of the aforementioned herbal extracts was evaluated using an ultra-high-pressure liquid chromatography (UHPLC) coupled with high-resolution mass spectrometry (HRMS) method to check the impact of the different preparations on the most abundant and significant compounds. All the extracts exerted a mild diuretic action, TCT and OpTC inducing the most intense diuretic effect. Both herbal preparations produced a statistically significant, dose-dependent and gradual increase of the urine output, the effect being more intense at 24h (6.63-7.13 ml/24h). Potentiometric evaluation of urine samples collected from treated rats revealed a clear and mild natriuretic and kaliuretic effect after the administration. In terms of antimicrobial activity, E. coli (MIC-0.38mg/ml), B. cereus (MIC-0.75mg/ml)), Penicillium funiculosum and P. verrucosum var. cyclopium (MIC-0.19mg/ml) showed the greater sensitivity to the tested extracts, respectively. UHPLC-HRMS screening showed that the bioactive potential of T. comosus herbal preparations was likely related to the higher amounts of phenolic acids (including rosmarinic acid), flavonoids (mainly flavones and derivatives) and other phenolics (such as different isomers of salvianolic acids) in their composition. The obtained results support the ethnopharmacological evidence regarding the mild diuretic and antibacterial potentials of the endemic wild thyme T. comosus, this study being the first one that assessed the aforementioned bioactivities for this species.

In Vivo and in vitro antitumor activity of tomatine in hepatocellular carcinoma

Background: There is abundant ethnopharmacological evidence the uses of regarding Solanum species as antitumor and anticancer agents. Glycoalkaloids are among the molecules with antiproliferative activity reported in these species. Purpose: To evaluate the anticancer effect of the Solanum glycoalkaloid tomatine in hepatocellular carcinoma (HCC) in vitro (HepG2 cells) and in vivo models. Methods: The resazurin reduction assay was performed to detect the effect of tomatine on cell viability in human HepG2 cell lines. Programmed cell death was investigated by means of cellular apoptosis assays using Annexin V. The expression of cancer related proteins was detected by Western blotting (WB). Reactive oxygen species (ROS) and calcium were determined by 2,7-dichlorodihydrofluorescein diacetate and Fluo-4, respectively. Intrahepatic HepG2 xenograft mouse model was used to elucidate the effect of tomatine on tumor growth in vivo. Results and Discussion: Tomatine reduced HepG2 cell viability and induced the early apoptosis phase of cell death, consistently with caspase-3, -7, Bcl-2 family, and P53 proteins activation. Furthermore, tomatine increased intracellular ROS and cytosolic Ca+2 levels. Moreover, the NSG mouse xenograft model showed that treating mice with tomatine inhibited HepG2 tumor growth. Conclusion: Tomatine inhibits in vitro and in vivo HCC tumorigenesis in part via modulation of p53, Ca+2, and ROS signalling. Thus, the results suggest the potential cancer therapeutic use of tomatine in HCC patients.

Review of Ethnobotanical and Ethnopharmacological Evidence of Some Ethiopian Medicinal Plants Traditionally Used for Peptic Ulcer Disease Treatment.

A peptic ulcer is described as the rupture of the mucosal integrity of the stomach, the duodenum, and, in certain cases, the lower esophagus as a result of contact with chloridopeptic secretions. The two most common kinds of peptic ulcer disorders are referred to as “gastric ulcer” and “duodenal ulcer.” The name is derived from the location of the ulceration. Despite the promise of a wide range of antiulcer treatments, these therapies are associated with several adverse reactions, including hypersensitivity, arrhythmia, impotence, gynecomastia, galactorrhea, hematological abnormalities, and kidney disease, which are intolerable for many patients. Nowadays, there is a lot of emphasis on finding new and innovative agents. As a result, herbal medicines are commonly utilized in circumstances when drugs are used for long periods and are also cost-efficient, effective, and readily available. In this review paper, a total of 82 medicinal plants have been identified and reported for their use in the treatment of peptic ulcer disease. The majority of these medicinal plants are widely used throughout Ethiopia. However, only the safety and efficacy of Plantago lanceolata, Osyris quadripartita, Rumex nepalensis, Cordia africana, Croton macrostachyus, and Urtica simensis have been scientifically studied in animal models. Despite this, many medicinal plants’ pharmacological effects and chemistry have not been well studied scientifically. As a result, further bioactive compound characterization, efficacy, mechanism of action evaluation, and toxicity evaluation of medicinal plants should be carried out. A study that can improve the documentation of indigenous knowledge and contribute to drug development and future self-reliance is also recommended.

Ethnobotanical Survey on Bitter Tea in Taiwan.

Ethnopharmacological evidence: In Taiwan, herbal tea is considered a traditional medicine and has been consumed for hundreds of years. In contrast to regular tea, herbal teas are prepared using plants other than the regular tea plant, Camellia sinensis (L.) Kuntze. Bitter tea (kǔ-chá), a series of herbal teas prepared in response to common diseases in Taiwan, is often made from local Taiwanese plants. However, the raw materials and formulations have been kept secret and verbally passed down by store owners across generations without a fixed recipe, and the constituent plant materials have not been disclosed. Aim of the study: The aim was to determine the herbal composition of bitter tea sold in Taiwan, which can facilitate further studies on pharmacological applications and conserve cultural resources. Materials and methods: Interviews were conducted through a semi-structured questionnaire. The surveyed respondents were traditional sellers of traditional herbal tea. The relevant literature was collated for a systematic analysis of the composition, characteristics, and traditional and modern applications of the plant materials used in bitter tea. We also conducted an association analysis of the composition of Taiwanese bitter tea with green herb tea (qing-cao-cha tea), another commonly consumed herbal tea in Taiwan, as well as herbal teas in neighboring areas outside Taiwan. Results: After visiting a total of 59 stores, we identified 32 bitter tea formulations and 73 plant materials. Asteraceae was the most commonly used family, and most stores used whole plants. According to a network analysis of nine plant materials used in high frequency as drug pairs, Tithonia diversifolia and Ajuga nipponensis were found to be the core plant materials used in Taiwanese bitter tea. Conclusion: Plant materials used in Taiwanese bitter tea were distinct, with multiple therapeutic functions. Further research is required to clarify their efficacy and mechanisms.

SYNERGISTIC IMMUNOMODULATORY ACTIVITY OF AQUEOUS ROOT EXTRACT OF ASPARAGUS RACEMOSUS WILLD AND ETHANOL WHOLE PLANT EXTRACT OF BOERHAVIA DIFFUSA LINN

Objective: On the basis of traditional use and Ethno pharmacological evidences Boerhavia diffusa whole plant and root part of Asparagus racemosus (Shatavari) both are widely used to enhance the immunity. But in combination there is no scientific evidence so current study was designed. Methods: Laboratory based study, namely, carbon clearance, cyclophosphamide induced immune suppression and neutrophil adhesion was designed using mice as an experimental animals in different combination of aqueous whole plant extract of B. diffusa and methanol extract of root part of A. racemosus were used as a test drug in the ratio 1:1, 1:2, and 2:1 (100 mg/kg) against the established standard drug Ashwagandha. Results: The results revealed that animals treated with combined extract (1:1, 1:2, and 2:1) at a dose of 100 mg/kg increase rate of carbon clearance from blood, there is significance alternation in blood parameter in cyclophosphamide group and also improve the Neutrophil adhesion when treated with different combination of polyherbal formulation treated groups. Conclusion: The polyherbal formulation in different ratio showing good significant immunomodulatory activity as compare to standard.

The toxicity of Aspidosperma subincanum to MCF7 cells is related to modulation of oxidative status and proinflammatory pathways

Ethnopharmacological relevanceCancer is an inflammatory disease because carcinogenesis and tumor progression depend on intrinsic and extrinsic inflammatory pathways. Although species of the genus Aspidosperma are widely used to treat tumors, and there is ethnopharmacological evidence for traditional use of the species A. subincanum as an anti-inflammatory agent, its antineoplastic potential is unknown. Aim of the studyTo evaluate toxic effects of the indole alkaloid–rich fraction (IAF) of A. subincanum on the MCF7 cell line and identify some of the anti-inflammatory mechanisms involved. Materials and methodsChromatographic analyses were performed by ultra-high-performance liquid chromatography with electrospray ionization mass spectrometry, and cytotoxic and antiproliferative effects of IAF were verified by MTT and clonogenic assays. Cell cycle alterations were analyzed by measuring DNA content, while propidium iodide and acridine orange staining was performed to determine the type of induced cell death. The expression of apoptosis markers and proteins involved in cell proliferation and survival pathways was analyzed by immunoblotting, RT-qPCR, and ELISAs. Interference with redox status was investigated using a DCFH-DA probe and by measuring catalase activity. ResultsChromatographic analyses showed that IAF is a complex mixture containing indole alkaloids. IAF selectively exerted toxic and antiproliferative effects, elevating the Bax/Bcl-xL ratio and inducing apoptosis in MCF7 cells. IAF decreased intracellular reactive oxygen species levels and increased catalase activity, while reducing the IL-8 level and suppressing COX-2 expression. ConclusionsIAF induces apoptosis in MCF7 cells by suppressing COX-2 expression while reducing IL-8 levels and intracellular content of reactive oxygen species.

Chemical Composition, Diuretic, and Antityrosinase Activity of Traditionally Used Romanian Cerasorum stipites.

Cherry stems (CS) represent a by-product intensively used in Eastern European countries as a traditional remedy for urinary tract disorders. Ethnopharmacological evidences sustain the use of CS as aqueous preparations (infusion and decoction), but few data were previously reported about phytochemical profile and pharmacological potential of CS hydroalcoholic extracts. In this regard, we aimed to evaluate the phenolic profile, in vitro antioxidant and tyrosinase inhibitory potential, and in vivo diuretic activity of 70% hydroethanolic cherry stems extract and cherry stems decoction (CSD). LC-DAD-ESI/MSn analysis revealed the presence of flavonoid-type compounds as main constituents for both preparations, especially flavanones (naringenin glycosides). Antioxidant activity evaluated through DPPH, ABTS, and FRAP methods was superior for cherry stems extract, probably due to the presence of phenolic-derived compounds in higher amounts than CSD. On the other hand, tyrosinase inhibitory potential and diuretic effect exerted by CSD were stronger, highlighting that other types of hydrophilic secondary metabolites are responsible for this bioactivity. Overall, our findings indicate that CS preparations could be used as promising mild diuretic agents and encourage further investigations regarding the correlation between their chemical composition and bioactive potential.

Quality assessment of ointment incorporated with Piper sarmentosum (Kaduk) leaves extract

Plant based medication is known as one of the alternative methods in wound management with no or less side effects, as well as less expensive. Various products related to wounds are on the market mostly consist of various unnatural chemicals. Previous preliminary study and ethnopharmacological evidence of Piper sarmentosum (Kaduk) also showed that the plant is a good candidate to treat wounds. Thus, this project aims to evaluate an ointment incorporated with the P. sarmentosum aqueous extract as an alternative to treat wounds. Two type of ointments; hydrocarbon (oleaginous; ointment 1) and water removeable (oil-in-water; ointment 2) that incorporated with the plant extract were further assessed for its organoleptic, physicochemical properties, as well as its stability at different storage conditions. The findings showed that ointment 1 exhibited good physicochemical properties with a strong scent of extract. However, this ointment did not inhibit E. coli, S. aureus and B. subtilis in the antimicrobial test. The stability study that carried out for three weeks consecutively showed that ointment 1 (hydrophilic) was stable under chill storage condition (5 °C) and room temperature (25 °C) without phase separation, as well as insignificant change of odour, texture and pH value. However, the formulation was physically unstable at high temperature (45 °C). Meanwhile, ointment 2 (oil-in-water) significantly inhibited E. coli, S. aureus and B. subtilis possibly due to presence of parabens in the formulation., not melted at high temperature (45 °C), stable in pH, non-greasy and water-washable. However, the formulation was physically unstable, that it showed phase separation after heating, and harden in texture when compared to the commercial healing ointment (Brand: Zam Buk). Findings from this study could be an initial step for the future development of the effective, affordable and safe wound healing medication by related industries.

A Review on Phytopharmaceuticals having Concomitant Experimental Anti-diabetic and Anti-cancer Effects as Potential Sources for Targeted Therapies Against Insulin-mediated Breast Cancer Cell Invasion and Migration

Diabetes and breast cancer are pathophysiologically similar and clinically established diseases that co-exist with a wider complex similar molecular signalling and having a similar set of risk factors. Insulin plays a pivotal role in the invasion and migration of breast cancer cells. Several ethnopharmacological evidences shed light on the concomitant anti-diabetic and anti-cancer activity of medicinal plant and phytochemicals against breast tumors of patients with diabetes. This present article reviewed the findings on medicinal plants and phytochemicals with concomitant antidiabetic and anti-cancer effects reported in scientific literature to facilitate the development of dual- acting therapies against diabetes and breast cancer. The schematic tabular form of published literature on medicinal plants (63 plants belongs to 45 families) concluded the dynamics of phytochemicals against diabetes and breast tumors that could be explored further for the discovery of therapies for controlling of breast cancer cell invasion and migration in patients with diabetes.

Withania somnifera in Neurological Disorders: Ethnopharmacological Evidence, Mechanism of Action and its Progress in Delivery Systems.

The underlying cause of major neurodegenerative disorders remains a healthcare mystery. The thoroughly investigated causes include oxidative stress, inflammation, environmental factor, mitochondrial dysfunction, and irregular neuronal protein aggregation. Withania somnifera has been used for more than 2500 years as a useful medicinal plant to improve disease defense, prevent aging, rejuvenate the body in a vulnerable situation, and generate a feeling of mental well-being. However, a persuasive paper emphasizing its neuroprotective nature is missing. In the current review, we have delineated the protective role of W. somnifera against various neurological disorders and its progress in delivery systems. The database used in the retrieval of data were PubMed, Scopus, Science direct, and SciFinder. The keywords used were W. somnifera, Ashwagandha, neuroprotective activities, etc. The principal source of the data retrieval includes research articles, review papers, and short communications from reputed publishers, including the New England Journal of Medicine, Elsevier, Nature, Springer, and Taylor & Francis. After an extensive literature review, we found that W. somnifera mitigates various neurological disorders, including Parkinson's disease, Alzheimer's disease, Huntington disease, tardive dyskinesia, stroke, and anxiety. Furthermore, natural compounds in nano sizes range possess better neuroprotective activity. Consequently, polymeric nanomicelles, nanoparticles, and nanofibers of natural products are used in the treatment of neurodegenerative diseases. The current review substantially deciphered the protective role of W. somnifera against various neurological disorders. However, future studies are further required better to understand the molecular mechanisms behind their neuroprotective nature.

Bioactivities of Colocasia affinis Schott and Profiling of its Bioactive Polyphenols by HPLC-DAD

Traditional medicines including Ayurveda, Unani have been used as therapeutics in the Indian subcontinent since 5000 BC. Because of the abundance of secondary metabolites, medicinal plants are becoming priceless natural resources for better drug development. Till date almost no ethnopharmacological evidences were available on Colocasia genus except Colocasia esculenta. As a part of our continuing assessment of evidence based use of traditional medicine, we have identified bioactive polyphenols in the ethanolic leaves extract of Colocasia affinis Schott using HPLC-DAD method and evaluated its anti-allergy, anti-inflammatory and anti-diabetic potentials for the first time. Chromatographic investigation showed the presence of p-coumaric acid, trans-ferulic acid, rosmarinic acid, myricetin and kaempferol in the leaves extract. Pharmacological profile exploration demonstrated its anti-inflammatory activity at a dose of 250- and 500-mg/kg in xylene-induced ear edema in mice. The extract significantly reduced the allergy like symptoms such as sneezing, scratching, rhinorrhea, redness and swelling in Tolune diisocyanate (TDI) sensitized allergy model mice at 250- and 500-mg/kg doses. Besides, the extract also exhibited reduction in the level of neutrophils, lymphocytes and eosinophils in a dose dependent manner in the experimental animal. The extract also confirmed reduction of blood glucose levels in the experimental mice at a dose of 250- and 500-mg/kg. Therefore, it is evident that C. affinis demonstrated possesses medicinal values through its anti-inflammatory, anti-allergic and anti-hyperglycaemic potentials. Bangladesh Pharmaceutical Journal 24(1): 1-10, 2021

Upregulation of Cyp19a1 and PPAR-γ in ovarian steroidogenic pathway by Ficus religiosa: A potential cure for polycystic ovary syndrome

Ethnopharmacological relevanceQuite a few plants are in use to treat female infertility and associated problems. Availing the cues from traditional knowledge, phytochemical studies and ethnopharmacological evidences, the aphrodisiac plant Ficus religiosa (F. religiosa) is widely in use to cure infertility in women. For instance, the juice of leaf and aerial root of F. religiosa is reported to normalize the dysregulated menstrual cycle in women. Besides, it is believed that regular circumambulation of F. religiosa during the early hours of the morning helps women in alleviating infertility which could be attributed to the potential phytovolatiles released from F. religiosa. However, the evidences for therapeutic potential of F. religiosa in treating female infertility are arbitrary and mostly anecdotal. Aim of the studyThe present study was aimed at examining if extracts of fresh and/or dry leaf of F. religiosa would cure polycystic ovary syndrome (PCOS) in the rat model. MethodsRats were divided into seven groups; control (Group I), PCOS-induced (P.O, Letrozole −1 mg/kg BW for 21 days) and untreated (Group II), PCOS-induced and treated with the leaf extracts of F. religiosa (Groups III–VI), and, PCOS-induced and treated with pioglitazone (Group VII). The estrous intervals, body and organ weights (ovary and uterus), and serum hormones (testosterone, luteinizing hormone [LH], estrogen, and progesterone) were measured, and the expression of Cyp19a1 (aromatase), and Peroxisome Proliferator-Activated Receptor-γ (PPAR-γ) were assessed in the experimental rats. The levels of 3β-hydroxysteroid dehydrogenase (3β-HSD), 17β-hydroxysteroid dehydrogenase (17β-HSD), and antioxidants (MDA, GSH, GPx, SOD, and CAT) were also quantified. Besides, the putative volatile compounds in the esterified leaf extracts were identified using Gas Chromatography-Mass Spectrometry (GC-MS). ResultsLetrozole treatment induced irregular estrous and altered weight of organs and hormonal milieu, which were reverted to normal in leaf extracts-treated PCOS-induced rats. Remarkably, fresh leaf treatment up-regulated Cyp19a1and PPAR-γ and increased the levels of 3β-HSD and 17β-HSD. We found 3-acetoxy-3-hydroxy-propionic acid in fresh and dry leaf extracts, which we attribute to efficacy of the extracts in alleviating PCOS. ConclusionPut together, our findings suggest the leaves of F. religiosa as potential in alleviating PCOS, mainly due to the presence of putative volatile molecules. Further screening of the leaves of F. religiosa is recommended to identify other key molecules and to develop a systematic therapeutic intervention for PCOS.

Anti-inflammatory activities of Canarium subulatum Guillaumin methanol extract operate by targeting Src and Syk in the NF-κB pathway

Ethnopharmacological relevanceCanarium subulatum Guillaumin is an herbal medicinal plant native to Southeast Asia. Ethnopharmacological evidence suggests that plants of the genus Canarium cure a variety of inflammatory diseases. Aim of the studyThe pharmacological mechanisms of C. subulatum Guillaumin remain poorly understood. In this study, we investigate inflammatory mechanisms and target molecules using C. subulatum Guillaumin methanol extract (Cs-ME) in inflammatory reactions managed by macrophages. Materials and methodsTo identify the anti-inflammatory activities of Cs-ME, lipopolysaccharide (LPS)-stimulated macrophages and a murine HCl/EtOH-induced gastritis model were chosen. The luciferase reporter gene assay, Western blot analysis, overexpression strategy, and the cellular thermal shift assay (CETSA) were employed to investigate the molecular mechanisms and target enzymes of Cs-ME. The active ingredients of this extract were also determined by HPLC. ResultsReleased levels of nitric oxide (NO) and mRNA expression levels of iNOS and IL-6 were downregulated by Cs-ME without exhibiting cytotoxicity. This extract inhibited MyD88-induced promoter activity and the nuclear translocation of nuclear factor (NF)-κB. Moreover, we found that Cs-ME reduced the phosphorylation of NF-κB upstream signaling molecules including IκBα, IKKα/β, Src, and Syk in LPS-stimulated macrophage-like RAW264.7 cells. The results of Western blot and CETSA confirmed that Src and Syk are anti-inflammatory targets of Cs-ME. In addition, orally injected Cs-ME alleviated HCl/EtOH-induced gastric ulcers in mice. HPLC analysis indicated that quercetin, luteolin, and kaempferol are major active components of this extract with anti-inflammatory activity. ConclusionsCs-ME exhibits anti-inflammatory effects in vitro and in vivo by targeting Src and Syk in the NF-κB signaling pathway. Consequently, Cs-ME could be developed as an anti-inflammatory herbal medicine.

Ligustilide Targets SMAD3, Showing Immunomodulatory Effects in Improving Atherosclerosis

Backgrounds: Atherosclerosis is initiated by the local immune response to lipid deposition, and the most commonly administered antiatherogenic drugs are lipid regulators, particularly statins. However, statins appear to be weak in regulating immunity. Based on traditional ethnopharmacological evidence, in this paper, we aimed to find effective therapeutic agents other than statins and to elucidate their potential action mechanisms. Methods: A traditional Chinese Medicine (TCM), Suxiao Jiuxin Pill (SX) has been widely used in curing cardiovascular diseases for thirty years. With a combination of pharmacologic studies on atherosclerosis and RNA-Seq transcriptomics were employed to explore the pharmacodynamic differences of SX and atorvastatin in the ApoE-/- mouse. The active ingredients in SX, were screened by biomarker CD137, which is closely associated with anti-atherosclerosis. Then, chemical proteomics based on a ligustilide-derived photoaffinity probe were applied to identify potential targets and active mechanism for developing the therapeutic effects of SX. Findings: Differentially expressed genes (DEGs) that were modulated by SX to a greater degree than atorvastatin were primarily involved in immunomodulation, and secondarily in cholesterol export. Ligustlide was the active ingredient in SX and its potential target was SMAD3. Ligustilide suppressed the phosphorylation and nuclear translocation of SMAD3 by blocking the mutual interaction of SMAD3 and transforming growth factor-β (TGF-β) receptor І. Interpretation: Ligustilide suppresses local lipid accumulation and achieves immunomodulation by targeting SMAD3 and the TGF-β1/SMAD3 pathway, thereby improving atherosclerosis. Developing a novel SMAD3 inhibitor may be a therapeutic option for preventing atherosclerosis in combination therapy with statins Funding Statement: This work was financially supported by National Key Research and Development Program of China (No. 2018YFC1704805, 2018YFC1704505) and International Cooperation and Exchange of the National Natural Science Foundation of China (No. 81761168039). Declaration of Interests: The authors declare no conflict of interest. Ethics Approval Statement: All animal procedures conformed to the Council of Europe Convention Directive (2010/63/EU) for the protection of vertebrate animals used for scientific purposes. And all animal procedures were performed strictly according to the guidelines for the care and use of laboratory animals, which was approved by Tianjin University of Traditional Chinese Medicine (TCM-LAEC2016032).