Major Bioactive Constituents Research Articles

Croton cajucara: Patents and Nanotechnological Advances.

Croton cajucara Benth showed several pharmacological properties such as: antiinflammatory, antinociceptive hypoglycemic, lipid-lowering, antiulcer, antiestrogenic, antitumor, antigenotoxic, antimutagenic, and cardiovascular. The 19-nor-clerodane diterpene transdehydrocrotonin (t-DCTN or DCTN) is the major bioactive constituent extracted from the bark of this Croton. Patents for Croton cajucara Benth in the period 2015 to 2022 comprises 14 published documents. Among them 4 patents are colloidal systems (SM/SNEDDS) loading t-DCTN for pharmacological applications. Patent registrations highlighted the huge promising biotechnological potential of Croton cajucara Benth especially in the phytotherapy field, and the correlation with its bioactive constituents of which t-DCTN showed the foremost results, so this herbal could become an alternative in the treatment of COVID-19. However, investigation of more recently published patents for clerodane diterpenes with similar chemical structure of t-DCTN, who previously showed antiviral property must be carried out and should be searched on several patent data bases.

Changes of polyphenols and their antioxidant activities in non-pigmented, red and black rice during in vitro digestion

Polyphenols, a major bioactive constituent in rice grain, require processing and digestion before being absorbed by human body. Free and bound phenolics, flavonoids and their antioxidant activities in non-pigmented, red and black rice after cooking and INFOGEST digestions of oral, gastric and intestinal phases were investigated. It showed that cooking caused great losses of polyphenols and antioxidant activity. Free ferulic, isoferulic and p-coumaric acid in most rice were highest at intestinal phase (p < 0.05). Bound ferulic acid in three colored rice, bound p-coumaric acid in black rice and catechin in red rice were higher at oral and/or gastric phase. After cooking, total flavonoids of non-pigmented and pigmented rice were highest at intestinal and gastric phase, respectively. Cyanidin-3-O-glucoside, peonidin-3-O-glucoside and quercetin peaked at intestinal phase in black rice. It suggested that black rice has a greater potential to be used in meal balance and functional product development.

Geniposide attenuates influenza virus-induced pneumonia by regulating inflammatory cytokines production. Evidences to elucidate the followed pathway

BackgroundInfluenza virus-induced pneumonia (IVP) is an infectious pulmonary disease characterized by exacerbated pulmonary inflammation caused by invasion of the influenza virus. IVP continues to threaten public health due to its high morbidity and mortality rates. Geniposide is one of the major bioactive constituents of G. jasminoides, which exerts antiviral and anti-inflammatory effects on influenza A virus (IAV) infection. PurposeTo investigate therapeutic effects and comprehensive mechanisms of geniposide on IAV infection and subsequent pneumonia. MethodsICR mice were infected intranasally with H1N1 (A/FM/1/47) to detect the anti-IAV activity of geniposide. Proteomics combined with function-integrated analysis were conducted to gain insight into the comprehensive mechanisms of geniposide. Subsequently, western blot was used to detect the phosphorylation of signal transducer and activator of transcription 1 (STAT1), signal transducer and activator of transcription 2 (STAT2), Interferon regulatory factor 9 (IRF9) and Janus kinase 1 (JAK1) in Janus kinase/signal transducer and activator of transcription (JAK/STAT) signaling pathway in lung tissue. Finally, RT-qPCR was used to detect the levels of interleukin 6 (IL-6), interleukin 17 (IL-17), interferon-γ (IFN-γ) and the STAT1 inhibitor (fludarabine) was used to verify the targeting between STAT1 and geniposide in RAW cells. ResultsGeniposide could significantly reduce the lung index, diminish lung pathology, decrease the virus loads and the inflammatory cytokines expression induced by IAV infection. A total of 411 differentially expressed proteins were identified among control, model, and geniposide-treated group in proteomic analysis. According to function-integrated analysis, 15 KEGG pathways were enriched and divided into 9 groups (modules), including influenza A, NOD-like receptor signaling, RIG-I-like receptor signaling, and so on. Among these modules, the most intensely interacting module pair was the NOD-like receptor signaling and influenza A, in which STAT1 and STAT2 acted as hubs with critical bridgeness role in the target network of geniposide. This indicated that geniposide may mitigate inflammation and alleviate IVP by JAK/STAT signaling pathways. Moreover, validation experiments confirmed that geniposide can significantly inhibit STAT1 and STAT2 phosphorylation as well as down-regulated expression of IL-6, IFN-γ and IL-17 in lung. Furthermore, when RAW cells were treated with the STAT1 inhibitor (fludarabine), the inhibitory effect of geniposide on IFN-γ and IL-6 was attenuated significantly. ConclusionsGeniposide can attenuate IAV-induced pneumonia by regulating inflammatory cytokines production through the JAK/STAT pathway.

Preclinical Toxicity Assessment of Polyphenols-Based Standardized Extract of Cinnamomum zeylanicum Bark

Background: Cinnamon (Cinnamomum zeylanicum) bark is used as a spice in culinary practices and has been documented for health and medicinal benefits. Polyphenol (PP) is the major bioactive constituent of cinnamon bark. However, acceptable and safe dose levels and toxicity information of oral exposure to PP-based standardized cinnamon bark extract (PP-CZ) are crucial for safe human consumption. Objective: To evaluate PP-CZ for acute oral, subchronic oral toxicities in vivo, mutagenicity in vitro and genotoxic potential in vitro. Method:. The effects of oral treatment with single- and 90-days repeated dose were evaluated in rates as per OECD Test 423 and 408, respectively. Effects on body weight, food and water intake, organ weight, hematology, biochemistry, and histology were recorded . Mutagenicity and genotoxicity were evaluated using Ames (OECD No. 471) and chromosomal aberrations (OECD Test. 473) tests. Results: A single oral dose of PP-CZ did not cause death or treatment-related toxic effects, indicating a “median lethal dose” &gt; 2,000 mg/kg. In addition, a subchronic dosage (500 mg/kg/day, 90 days) was found safe in rats, suggesting “no observed adverse effect level” (NOAEL) of 500 mg/kg and “Human Equivalent Dose” (HED) of 4.8 g/day. Furthermore, the absence of mutagenicity or genotoxicity of PP-CZ was observed during in vitro tests. PP-CZ showed a robust safety profile without mutagenicity or genotoxicity in rats.

Embelin Novel Drug Molecule: A Review

Embelia ribes Burm family (Myrsinaceae), an important traditional medicinal plant of India. Embelia ribes have been proven to have great pharmacological potential with great utility and usage as traditional medicine. The root, berries, and leaves of Embelia ribes are used in herbal formulations. E. ribes fruits contain a quinone derivative (2, 5-dihydroxy-3-undecyl-1, 4-benzoquinone), Embelin which is the major bioactive constituent. It has a wide spectrum of biological activities, including antioxidant, antitumor, anti-inflammatory, analgesic, anthelmintic, antifertility, antimicrobial, antibacterial, and antiprotozoal in abdominal disorders, lung diseases, constipation, indigestion, fungal infections, mouth ulcer, sore throat, pneumonia, heart disease, and obesity. Embelin was isolated by cold and hot extraction with polar and nonpolar solvents. The concentrated and dried extracts were washed with petroleum ether to get crystalline embelin. Isolated embelin was characterized by melting point, TLC, partition coefficient, solubility, HPLC, HPTLC, FT-IR, NMR, and mass spectroscopy.

Phytochemical and Pharmacological Profile of Desmodium gangeticum (L.) DC.: A Comprehensive Review

Abstract: Desmodium gangeticum (L.) DC., commonly known as Shalparni in Ayurveda, has been used for treating various diseases. The plant is one among ten ingredients of classical Ayurvedic formulations Dasamula. Decoctions of root and leaf, root powder and leaf juice of D. gangeticum are used as tonics, febrifuge, digestive, antiemetic, astringent, anti-asthmatic, anti-diarrhoeal and anti-inflammatory agents in traditional medicinal practices of India and its neighbouring countries. Phytochemical investigations of this plant revealed the presence of many bioactive phytoconstituents. The present review was prepared with the objective of documenting the phytochemical and pharmacological profiles of D. gangeticum. The data presented here were collected from various sources like books, journals, various online databases, and monographs and internet search engines Google Scholar, PubMed, Science Direct, etc. Studies reported from phytochemical screening revealed that D. Gangeticum leaves and roots are rich in flavonoids, alkaloids, terpenoids, tannins, phenols, carbohydrates, steroids, phenylpropanoids, pterocarpans, coumarins, and volatile oil. Among the isolated compounds, major bio-active constituents are alkaloids, flavonoids and pterocarpans. All these phytochemicals are widely distributed in various parts of D. gangeticum. The other active constituents like desmodin, hordenine and gangetin are largely responsible for its broad spectrum of therapeutic potentiality. Desmodin belongs to the class of pterocarpans and is reported to have antifungal and antibacterial activities. Gangetin, which also belongs to the pterocarpans, is reported to have anti-inflammatory and analgesic effects. FTIR spectroscopy showed that D. gangeticum extracts are mainly rich in phenolic derivatives. Scientific experiments carried out in vitro as well as preclinical animal experimentations, showed that the plant and its bioactive chemicals exhibit various therapeutic activities. It is reported that D. gangeticum extracts showed potent antioxidant, antibacterial, hypoglycaemic, analgesic, hepatoprotective, anti-inflammatory and antiarthritic activities. Studies reported that D. gangeticum or its formulations are safe when used in appropriate therapeutic dosage.

Exploring the antioxidant and antidiabetic potentials of Syzygium cumini L. landraces: phytochemicals, bioactive constituents and pathway enrichment analysis

SummaryThis study proposes an efficient antidiabetic strategy by identifying potential mechanisms of antioxidant metabolism and α‐amylase inhibition in Syzygium cumini L. fruits. Six indigenous landraces of S. cumini were evaluated for major phytochemicals and bioactive constituents using gas chromatography–mass spectrometry (GC–MS) profiling and correlated with antioxidant and antidiabetic properties. Black jamin landrace‐6 (BJLR‐6), characterised by small fruits, exhibited greater 1,1‐diphenyl‐2‐picrylhydrazyl (DPPH) scavenging activity and in vitro α‐amylase inhibition, especially in the kernel. The kernel of BJLR‐6 had the lowest IC50 for DPPH radicals (4.2 μg mL−1) and α‐amylase inhibition (5.3 μg mL−1). Therapeutic compounds like terpenoids in the pulp and alkaloids in the seed contribute to their curative potential. GC–MS profiling of BJLR‐6 revealed major bioactive constituents like propanamide, allantoic acid and 2‐furancarboxaldehyde in the pulp and alkaloids such as piperazine, 3‐pyridinol, imidazole and furan carboxaldehyde in the kernel tissues. Pathway enrichment analysis of bioactive constituents emphasises antioxidant and insulin‐regulating pathways, steroid hormone metabolism and signal transduction pathways associated with therapeutic/antidiabetic activity in fruit pulp and kernel tissues. This study unveils the phytochemical composition and novel bioactive constituents in the fruit parts of small‐fruited S. cumini BJLR‐6, emphasising their potential as a better source of functional herbal formulations.

Plumbagin ameliorates LPS-induced acute lung injury by regulating PI3K/AKT/mTOR and Keap1-Nrf2/HO-1 signalling pathways.

Acute lung injury (ALI) is a major pathophysiological problem characterized by severe inflammation, resulting in high morbidity and mortality. Plumbagin (PL), a major bioactive constituent extracted from the traditional Chinese herb Plumbago zeylanica, has been shown to possess anti-inflammatory and antioxidant pharmacological activities. However, its protective effect on ALI has not been extensively studied. The objective of this study was to investigate the protective effect of PL against ALI induced by LPS and to elucidate its possible mechanisms both invivo and invitro. PL treatment significantly inhibited pathological injury, MPO activity, and the wet/dry ratio in lung tissues, and decreased the levels of inflammatory cells and inflammatory cytokines TNF-α, IL-1β, IL-6 in BALF induced by LPS. In addition, PL inhibited the activation of the PI3K/AKT/mTOR signalling pathway, increased the activity of antioxidant enzymes CAT, SOD, GSH and activated the Keap1/Nrf2/HO-1 signalling pathway during ALI induced by LPS. To further assess the association between the inhibitory effects of PL on ALI and the PI3K/AKT/mTOR and Keap1/Nrf2/HO-1 signalling, we pretreated RAW264.7 cells with 740Y-P and ML385. The results showed that the activation of PI3K/AKT/mTOR signalling reversed the protective effect of PL on inflammatory response induced by LPS. Moreover, the inhibitory effects of PL on the production of inflammatory cytokines induced by LPS also inhibited by downregulating Keap1/Nrf2/HO-1 signalling. In conclusion, the results indicate that the PL ameliorate LPS-induced ALI by regulating the PI3K/AKT/mTOR and Keap1-Nrf2/HO-1 signalling, which may provide a novel therapeutic perspective for PL in inhibiting ALI.

Angiogenic protein profiling, phytochemical screening and in silico anti-cancer targets validation of stem, leaves, fruit, and seeds of Calotropis procera in human liver and breast cancer cell lines

The main focus of anticancer drug discovery is on developing medications that are gentle on normal cells and should have the ability to target multiple anti-cancer pathways. Liver cancer is becoming a worldwide epidemic due to the highest occurring and reoccurring rate in some countries. Calotropis procera is a xerophytic herbal plant growing wildly in Saudi Arabia. Due to its anti-angiogenic and anticancer capabilities, “C. procera” is a viable option for developing innovative anticancer medicines. However, no study has been done previously, to discover angiogenic and anti-cancer targets which are regulated by C. procera in liver cancer. In this study, leaves, stems, flowers, and seeds of C. procera were used to prepare crude extracts and were fractionated into four solvents of diverse polarities. These bioactivity-guided solvent fractions helped to identify useful compounds with minimal side effects. The phytoconstituents present in the leaves and stem were identified by GC-MS. In silico studies were done to predict the anti-cancer targets by major bioactive constituents present in leaves and stem extracts. A human angiogenesis antibody array was performed to profile novel angiogenic targets. The results from this study showed that C. procera extracts are an ideal anti-cancer remedy with minimum toxicity to normal cells as revealed by zebrafish in vivo toxicity screening assays. The novel antiangiogenic and anticancer targets identified in this study could be explored to design medication against liver cancer.

Aged garlic extract preserves beta-cell functioning via modulation of nuclear factor kappa-B (NF-κB)/Toll-like receptor (TLR)-4 and sarco endoplasmic reticulum calcium ATPase (SERCA)/Ca2+ in diabetes mellitus

BackgroundPeripheral insulin resistance and compromised insulin secretion from pancreatic β-cells are significant factors and pathogenic hallmarks of diabetes mellitus (DM). NF-κβ/TLR-4 and SERCA/Ca2+ pathways have been identified as potential pathways regulating insulin synthesis by preserving pancreatic β-cell functioning. The current study aimed to evaluate the therapeutic effect of aged garlic extract (AGE) against DM in a streptozotocin (STZ)-induced rat model with particular emphasis on pancreatic β-cell functioning.MethodsAGE was characterized by gas chromatography-mass spectrometry (GC-MS), Fourier-transform infrared spectroscopy (FTIR), and scanning electron microscopy (SEM) to evaluate its physio-chemical characteristics followed by in-vitro anti-diabetic and antioxidant potential. This was followed by the induction of DM in laboratory animals for investigating the therapeutic action of AGE by evaluating the role of NF-κβ/TLR-4 and the SERCA/Ca2+ pathway. The parameters assessed in the present experimental setup encompassed antioxidant parameters, metabolic indicators, insulin concentration, intracellular calcium levels, apoptotic markers (CCK-8 and Caspase Glo-8), and protein expression (P-62 and APACHE-II).ResultsAGE characterization by SEM, GC-MS, and X-ray diffraction (XRD) revealed the presence of phenylalanine, alliin, S-allylmercaptocysteine (SAMC), tryptophan, 1-methyl-1,2,3,4-tetrahydro-β-carboline-3-carboxylic acid as major bioactive constituents of AGE. Metabolic studies, including intraperitoneal glucose tolerance test (IPGTT), revealed significantly lower blood glucose levels in the AGE group compared to the disease control group. In contrast, the intraperitoneal insulin tolerance test (ITT) exhibited no significant difference in insulin sensitivity between the AGE supplementation group and the DM control group. Interestingly, AGE was found to have no significant effect on fasting glucose and serum insulin levels. In contrast, AGE supplementation was found to cause significant hypoglycaemia in postprandial blood glucose and insulin levels. Importantly, AGE causes restoration of intracellular Ca2+ levels by modulation of SERCA/Ca2 functioning and inhibition NF-κB/TLR-4 pathway. AGE was found to interact with and inhibit the DR-5/ caspase-8/3 apoptotic complex. Furthermore, microscopic studies revealed degeneration and apoptotic changes in pancreatic β-cells of the DM control group, while supplementation of AGE resulted in inhibition of apoptotic pathway and regeneration of pancreatic β-cells.ConclusionThe current study suggests that AGE enhance glucose homeostasis by exerting their effects on pancreatic β-cells, without ameliorating peripheral sensitivity. Moreover, AGEs promote an increase in β-cell mass by mitigating the apoptosis of pancreatic β-cells. These findings suggest that AGE could aid in developing a viable alternative therapy for diabetes mellitus (DM).

Integrated metabolomic and transcriptomic analysis of triterpenoid accumulation in the roots of Codonopsis pilosula var. modesta (Nannf.) L.T.Shen at different altitudes.

Codonopsis Radix is a beneficial traditional Chinese medicine, and triterpenoid are the major bioactive constituents. Codonopsis pilosula var. modesta (Nannf.) L.T.Shen (CPM) is a precious variety of Codonopsis Radix, which is distributed at high mountain areas. The environment plays an important role in the synthesis and metabolism of active ingredients in medicinal plants, but there is no report elaborating on the effect of altitude on terpenoid metabolites accumulation in CPM. This study aims to analyse the effects of altitude on triterpenoid biosynthetic pathways and secondary metabolite accumulation in CPM. The untargeted metabolomics based on liquid chromatography-tandem mass spectrometry (LC-MS/MS) and 10 triterpenoids based on ultra-performance liquid chromatography quadrupole time-of-flight mass spectrometry (UPLC-Q-TOF-MS) method were analysed at the low-altitude (1480 m) and high-altitude (2300 m) CPM fresh roots. The transcriptome based on high-throughput sequencing technology were combined to analyse the different altitude CPM triterpenoid biosynthetic pathways. A total of 17,351 differentially expressed genes (DEGs) and 55 differentially accumulated metabolites (DAMs) were detected from the different altitude CPM, and there are significant differences in the content of the 10 triterpenoids. The results of transcriptome study showed that CPM could significantly up-regulate the gene expression levels of seven key enzymes in the triterpenoid biosynthetic pathway. The CPM at high altitude is more likely to accumulate triterpenes than those at low altitude, which was related to the up-regulation of the gene expression levels of seven key enzymes. These results expand our understanding of how altitude affects plant metabolite biosynthesis.

Anti-cancer Potential of Phytoflavonoidal Drugs against Gynecological Cancer

Background: Flavonoids, one of the major bioactive constituents of herbal drugs, have been scientifically reported to possess diverse therapeutic potentials such as anticancer, immunomodulatory, neuroprotective, cardioprotective, antioxidant, etc. This manuscript enlightens the anticancer potential of traditional herbal flavonoids in gynecological cancer i.e., is one of the major life-threats in women. Objective: This manuscript is aimed at an insightful compilation of scientific substantiations of herbal flavonoids in gynecological cancer along with targeted drug delivery systems for the same. Materials and Methods: The contents and data represented in the article have been reviewed using institutional libraries and online database resources (available in the public domain) such as PubMed, Science-Direct, Web of Science, American Association of Pharmaceutical Scientists, Google Scholar, Hinari, SciFinder, Research Gate, etc. Results: Flavonoids are natural compounds and have potential against cervical, ovarian, and endometrial cancer. In-vitro and in-vivo experiments have demonstrated the significant potential of flavonoids in gynecological cancer, especially cervical, ovarian, and endometrial cancer. It was reported from in-vitro experimentations that targeted drug delivery system improves the anticancer effect of flavonoids. Conclusion: Phytoflavonoids have the potential to prevent gynecological cancer by induction of apoptosis cell cycle arrest and reactive oxygen species generation. Further studies on the drug delivery system of flavonoids are warranted.

Persicaria senticosa extract mitigates ultraviolet B-induced photoaging by suppressing the mitogen-activated protein kinase/activator protein 1/matrix metalloproteinase 1 pathway in human keratinocytes and hairless mice

Ultraviolet (UV) irradiation has been identified as a key trigger for skin photoaging, characterized by the overproduction of matrix metalloproteinases (MMPs) and reactive oxygen species (ROS), along with the accelerated decomposition of extracellular matrix (ECM) proteins, ultimately contributing to the development of wrinkles. Persicaria senticosa (PS) extracts are recognized for their antioxidative properties and their importance in skin health. Nevertheless, there is a paucity of studies investigating the potential of PS in protecting the skin against photoaging. The present study aimed to assess the effectiveness of PS extracts in preventing photoaging and elucidating the molecular mechanisms involved in using immortalized human keratinocytes (HaCaT) and hairless mice. The major bioactive constituents of PS were identified as p-coumaric acid, isoquercitrin, quercetin-3-O-glucuronide, and quercetin. Aqueous extracts of PS exhibited the ability to mitigate UVB-induced cellular damage and diminished ROS generation in HaCaT cells. Moreover, treatment with PS effectively attenuated the upregulated expression of matrix metalloproteinase-1 (MMP-1) and collagen degradation induced by UVB exposure. The property of PS to counteract photoaging was related to its capacity to inhibit the UVB-induced phosphorylation of mitogen-activated protein kinase (MAPK) and suppress the subsequent activation of activator protein 1 (AP-1) signaling pathways. Moreover, in hairless mice exposed to UVB radiation, the application of PS significantly alleviated the development of skin wrinkles, diminished epidermal thickening, and mitigated collagen degradation. Notably, PS treatment resulted in the downregulation of the UVB-activated MAPK/AP-1/MMP-1 pathway in mouse skin tissues. These findings suggest that PS has the potential to serve as a therapeutic agent for treating photoaging, holding promises in both cosmeceutical and pharmaceutical applications.

Catalpol alleviates amyloid- generation and neuronal oxidative stress injury via activating the Keap1-Nrf2/ARE signaling pathway in the immortalized lymphocytes from patients with late-onset Alzheimer's disease and SKNMC cells co-culture model.

To assess the effect of catalpol, the major bioactive constituents of Rehmannia glutinosa, on our Alzheimer's disease (AD) in vitro model. We employed the immortalized lymphocytes (lymphoblastoid cell line, LCL) from late-onset AD patients and co-cultured "them" to mimic the pathological process of late-onset AD and investigated the effect of catalpol on our AD in vitro model. In the co-culture model, AD-derived LCL triggered excessive Aβ1-42 in SKNMC cells due to its high levels of oxidative stress and resulted in neuronal oxidative stress injury through inhibiting Keap1-Nrf2/ARE signaling pathway. Treatment with catalpol and N-acetylcysteine (NAC), an antioxidant, prevented the AD LCL-induced Aβ1-42 overproduction and reduced the level of β-site amyloid precursor protein cleaving enzyme-1 (BACE1) and amyloid precursor protein (APP)-C99. Catalpol and NAC also enhanced the antioxidant capacity and reduced apoptosis in SKNMC cells co-cultured with AD LCL. The anti-oxidative effect of catalpol was antagonized by ML385, the Nrf2 inhibitor. Therefore, we speculate that the antioxidant and anti-apoptotic effects of catalpol are mediated by activating the Keap1-Nrf2/ARE signaling pathway. Catalpol affects the anti-Aβ generation and the antioxidative and antiapoptotic properties in the AD co-cultured model. So, it might be a novel natural drug and offer a potential therapeutic approach for AD.

Neocarya macrophylla (Chrysobalanaceae): Review of taxonomy, ethnobotany, phytochemistry and biological activities

The Neocarya macrophylla plant is used traditionally for food, medicinal, spiritual and industrial purposes. It is also used as a soap, dye, glue, fodder, termite repellent, firewood and for structural materials. Few studies on the physicochemical, nutritional content, phytochemical and pharmacological activities have validated the benefits of N. macrophylla to humanity as a food, in cosmetics or pharmaceutical products. The major bioactive constituents identified in the plant so far are steroids and flavonoids, such as, stigmasterol, quercetin, catechin and its related glycosides, as well as glucosamines. Extracts of the plant have shown good antivenom, antimicrobial, analgesic, anti-inflammatory, antimycobacterial, anthelmintic and antioxidant activities. Acute toxicity studies have confirmed the plant to be toxic. More studies on the plant are required in order to exploit other biological activities as claimed by traditional healers and also to isolate more bioactive compounds. In addition, the safety and tolerability assessment of N. macrophylla should be undertaken due to its widespread usage.

Crocetin inhibits choroidal neovascularization in both in vitro and in vivo models

Choroidal neovascularization (CNV) is the primary pathogenic process underlying wet age-related macular degeneration, leading to severe vision loss. Despite current anti-vascular endothelial growth factor (VEGF) therapies, several limitations persist. Crocetin, a major bioactive constituent of saffron, exhibits multiple pharmacological activities, yet its role and mechanism in CNV remain unclear. Here, we investigated the potential effects of crocetin on CNV using in vitro and in vivo models. In human umbilical vein endothelial cells, crocetin demonstrated inhibition of VEGF-induced cell proliferation, migration, and tube formation in vitro, as assessed by CCK-8 and EdU assays, transwell and scratch assays, and tube formation analysis. Additionally, crocetin suppressed choroidal sprouting in ex vivo experiments. In the human retinal pigment epithelium (RPE) cell line ARPE-19, crocetin attenuated cobalt chloride-induced hypoxic cell injury, as evidenced by CCK-8 assay. As evaluated by quantitative PCR and Western blot assay, it also reduced hypoxia-induced expression of VEGF and hypoxia-inducible factor 1α (HIF-1α), while enhancing zonula occludens-1 expression. In a laser-induced CNV mouse model, intravitreal administration of crocetin significantly reduced CNV size and suppressed elevated expressions of VEGF, HIF-1α, TNFα, IL-1β, and IL-6. Moreover, crocetin treatment attenuated the elevation of phospho-S6 in laser-induced CNV and hypoxia-induced RPE cells, suggesting its potential anti-angiogenic effects through antagonizing the mechanistic target of rapamycin complex 1 (mTORC1) signaling. Our findings indicate that crocetin may hold promise as an effective drug for the prevention and treatment of CNV.

Schisandrin B, a dual positive allosteric modulator of GABAA and glycine receptors, alleviates seizures in multiple mouse models.

Epilepsy is a prevalent and severe neurological disorder and approximately 30% of patients are resistant to existing medications. It is of utmost importance to develop alternative therapies to treat epilepsy. Schisandrin B (SchB) isa major bioactive constituent of Schisandra chinensis (Turcz.) Bailland has multiple neuroprotective effects, sedative and hypnotic activities.In this study, we investigated the antiseizure effect of SchB in various mouse models of seizure and explored the underlying mechanisms. Pentylenetetrazole (PTZ), strychnine (STR), and pilocarpine-induced mouse seizure models were established.We showed that injection ofSchB (10, 30, 60 mg/kg, i.p.) dose-dependentlydelayed the onset of generalized tonic-clonic seizures (GTCS), reduced the incidence of GTCS and mortality in PTZ and STR models. Meanwhile, injection ofSchB (30 mg/kg, i.p.) exhibited therapeutic potential in pilocarpine-induced status epilepticus model, which was considered as a drug-resistant model. In whole-cell recording from CHO/HEK-239 cells stably expressing recombinant human GABAA receptors (GABAARs) and glycine receptors (GlyRs) and cultured hippocampal neurons, co-application of SchB dose-dependently enhanced GABA or glycine-induced current with EC50 values at around 5 μM, and application of SchB (10 μM) alone did not activate the channels in the absence of GABA or glycine.Furthermore, SchB (10 μM) eliminated both PTZ-induced inhibition on GABA-induced current (IGABA) and strychnine (STR)-induced inhibition on glycine-induced current (Iglycine).Moreover, SchB(10 μM) efficiently rescued the impaired GABAARs associated with genetic epilepsies. In addition,the homologous mutants in both GlyRs-α1(S267Q) and GABAARs-α1(S297Q)β2(N289S)γ2L receptors by site-directed mutagenesis testsabolishedSchB-induced potentiation of IGABA and Iglycine. In conclusion, we have identified SchB as a natural positive allosteric modulator of GABAARs and GlyRs, supporting its potential as alternative therapies for epilepsy.

Pyrus pashia fruit extract and its major phytometabolite chrysin prevent hippocampal apoptosis and memory impairment in PTZ-kindled mice

ABSTRACT Objectives Epilepsy is a chronic neurological condition with recurrent seizures. One-third of epilepsy patients experience unacceptable side effects from antiepileptic drugs. Pyrus pashia is a deciduous tree from southern Asia. Ethnomedicinally, Malakand tribes use its fruits for epilepsy treatment. Our prior research demonstrated the anticonvulsive properties of ethanolic extract of Pyrus pashia (EPP) and its bioactive compound chrysin in acute seizure tests. This study aims to investigate the impact of EPP and chrysin on cognitive impairment in a PTZ-induced kindling mice model of epilepsy. Methods Swiss albino male mice were equally divided into four groups. The first group received 0.5% carboxy methyl cellulose dissolved in normal saline while the other three groups were pre-treated with Diazepam (DZP) (1 mg/kg, i.p.), EPP (200 mg/kg, p.o.) and chrysin (5 mg/kg, p.o.). After 30 min, all groups were administered PTZ (35 mg/kg, i.p.) and evaluated for seizure severity, cognitive function, and neuronal apoptosis. Western blot analysis was conducted to analyze the expressions of apoptosis biomarkers and memory-related genes, including cAMP response element-binding protein (CREB) and Brain Derived Neurotrophic Factor (BDNF). Results The therapeutic effects of EPP and Chrysin were comparable to DZP in terms of reducing seizure severity, but unlike DZP, they prevented PTZ-induced memory impairment in experimental animals. Additionally, they increased the levels of BDNF and CREB while reducing apoptotic biomarkers in the hippocampus of experimental animals. Conclusions Based on the leads offered by this study EPP and its major bioactive constituent, could be developed as the treatment option for epilepsy.

Full-length transcriptome sequencing provides new insights into the complexity of flavonoid biosynthesis in Glechoma longituba.

Glechoma longituba has been frequently used in treating urolithiasis and cholelithiasis due to the presence of flavonoids, which are its major bioactive constituents. However, research on the molecular background of flavonoid biosynthesis in G. longituba is limited. In this study, we used single-molecule real-time combined with next-generation sequencing technologies to construct the complete transcriptome of G. longituba. We identified 404,648 non-redundant transcripts, including 249,697 coding sequences, 197,811 simple sequence repeats, 174,846 long noncoding RNA, and 176,554 coding RNA. Moreover, we functionally annotated 346,218 isoforms (85.56%) and identified 86,528 differentially expressed genes. We also identified 55 non-redundant full-length isoforms related to the flavonoid biosynthetic pathway. Pearson correlation analysis revealed that the expression levels of some key genes of the flavonoid biosynthesis pathway were significantly positively correlated with the flavonoid metabolites. Furthermore, we performed bioinformatics analysis (sequence and structural) of isoform_47029 (encoding flavanone 3-hydroxylase) and isoform_53692 (encoding flavonol synthase) to evaluate their potential biological functions. Finally, we validated gene expression levels of 12 flavonoid-related key enzyme genes using quantitative real-time PCR. Overall, this study provides full-length transcriptome information on G. longituba for the first time and valuable molecular resources for further research on the medicinal properties of this plant.

Sesamol Alleviates High-Fat Diet-Induced Hepatic Insulin Resistance in C57BL/6J Mice Through AMPK Activation Mediated by Adipose Adiponectin.

Sesamol is the major bioactive constituent isolated from sesame seeds and has a variety of bioactivities. However, its role and mechanism in liver insulin resistance remain unknown. The current study was designed to investigate the underlying adipose-liver crosstalk mechanism of sesamol ameliorating hepatic insulin sensitivity. The therapeutic effect of sesamol was evaluated in high-fat diet (HFD)-fed C57BL/6J mice (100mg/kg for 8weeks, XYGW-2021-75) and the mechanism was further explored in HepG2 cells with/without adiponectin and adenosine 5 '-monophosphate-activated protein kinase (AMPK) inhibitor administration. Our in vivo data showed that sesamol reduced hepatic insulin resistance in HFD-induced mice with obesity by modulating protein expression levels of glycogen synthase (GS), phosphoenolpyruvate carboxykinase (PEPCK) and protein kinase B (AKT). Moreover, sesamol not only increased the serum and adipose tissue adiponectin concentrations but also activated the phosphorylation of AMPK in the liver. Furthermore, in vitro studies using recombinant human adiponectin and an AMPK inhibitor revealed that adiponectin and sesamol have a synergic impact on increasing glycogenesis and reducing gluconeogenesis, of which the effects could be attenuated by the AMPK inhibitor. Taken together, our results suggested that sesamol stimulated adiponectin secretion from adipocytes, whereby exhibited a co-effect on activating the downstream signal of hepatic AMPK, resulting in the alleviation of hepatic insulin resistance. The novel findings of sesamol on hepatic effects provides prospective therapeutic approaches to treat insulin resistance.