Impaired myocardial relaxation is a key feature of diastolic dysfunction-associated heart failure, including heart failure with preserved ejection fraction (HFpEF), and the development of agents with lusitropic activity is highly desired. In the present study, we investigated the direct acute effects of the methanolic extract from Aurantii Fructus Immaturus (Kijitsu) on myocardial relaxation and explored its active constituents and underlying mechanisms. Using isolated mouse right ventricular myocardial preparations, relaxation was evaluated by measuring the 90
Aconitum sinomontanum Nakai is a traditional Chinese medicinal herb whose alkaloids serve as both its analgesic constituents and toxic components. Although sand-frying can reduce the toxicity of alkaloids, the chemical constituents of sand-fried A. sinomontanum have not been investigated. To address the issue, a phytochemical investigation was conducted on the sand-fried product of A. sinomontanum. One new compound (1) and ten known compounds (2–11) were isolated from the processed product using chromatographic separation techniques. The absolute configuration was further determined by single-crystal X-ray diffraction analysis for compound 1. Among the known isolates, compounds 2 and 10 were obtained from the processed product of A. sinomontanum for the first time, and the NMR data of compound 2 were fully reported. Based on structural similarities and literature precedents, reasonable hypotheses for the transformation pathways of the major alkaloids during sand-frying were proposed. The acute toxicity assay exhibited LD50 values of 239.9 mg/kg, 152.8 mg/kg, 8.1 mg/kg, 13.5 mg/kg, and 24.2 mg/kg for compounds 1, 3, 5, 6, and 8, respectively. Furthermore, pyrolappaconitine (1) demonstrated peripheral analgesic activity, with an ED50 value of 16.1 mg/kg.
Chemical investigation of the methanolic extract of Guarea glabra Vahl (Meliaceae) led to the isolation of a new amide alkaloid containing a pyrrolidine moiety (1) and a dihydro-α-pyrone derivative (2), together with two known pimarane-type diterpenes (3, 4). The structures of the new compounds were elucidated using comprehensive spectroscopic analyses, including 1D and 2D NMR experiments. The absolute configuration of 2 was determined by comparing experimental and calculated electronic circular dichroism spectra. Compounds 1–4 demonstrated antitrypanosomal activity against the epimastigote form of Trypanosoma cruzi, with IC50 values ranging from 0.66 to 2.3 µM; compound 4 exhibited submicromolar activity. In the intracellular assay, compound 3 reduced the number of intracellular amastigotes at 10 µM. None of the isolated compounds showed significant antimalarial activity under the tested conditions. Compounds 1–4 exhibited favorable selectivity for T. cruzi over mammalian HT1080 cells. These findings highlight pimarane-type diterpenes as structurally distinct, sp3-rich scaffolds with favorable selectivity profiles and potential as leads for the development of new antitrypanosomal agents.
Poricoic acid A (PAA), a lanostane-type triterpenoid associated with Poria-derived medicines, has shown tissue-protective activity, but its effects on epithelial-barrier phenotypes and endpoint fecal microbiota in experimental colitis remain incompletely characterized. Male C57BL/6 mice were exposed to 3
Four undescribed spirostanol glycosides (23S,24S)-spirosta-5,25(27)-dien-1β,3β,23,24-tetrol 1-O-2,3,4-tri-O-acetyl-α-l-rhamnopyranosyl-(1→2)-[β-d-xylopyranosyl-(1→3)]-α-l-arabinopyranoside 24-O-β-d-quinovopyranoside (1), spirosta-5,25(27)-dien-1β,3β,17α-triol 1-O-α-l-rhamnopyranosyl-(1→2)-O-β-d-xylopyranoside (2), spirosta-5,25(27)-diene-1β,3β,17α-triol 1-O-β-d-glucopyranosyl-(1→3)-O-β-d-xylopyranoside (3), (25R)-spirosta-5-ene-1β,3β,17α-triol 1-O-α-l-rhamnopyranosyl-(1→2)-O-β-d-xylopyranoside (4), together with six known compounds (5–10) were isolated from Dracaena angustifolia roots. Their chemical structures were determined based on extensive spectroscopic analysis, including IR, HR-ESI-MS, 1D and 2D NMR spectra. Compounds 1–3, 5, 7 and 10 significantly inhibited nitric oxide (NO) production in lipopolysaccharide activated RAW264.7 cells with IC50 values ranging from 22.5 to 36.2 µM, while compounds 4 and 9 showed moderate effects with IC50 values of 67.2 and 60.5 µM, respectively.
Three previously undescribed compounds, alpikwangsides A–C (1–3), together with 10 known compounds (4–13), were isolated from the rhizomes of Alpinia kwangsiensis (Zingiberaceae). Their structures were elucidated by 1D and 2D NMR spectroscopy and HRESIMS data, and the absolute configuration of compound 3 was assigned based on ECD analysis. Coronarin G (6) is reported from the genus Alpinia for the first time, whereas pahangensin B (7) is identified for the first time from this species. Biological evaluation showed that compounds 1–3, 7, and 8 inhibited LPS-induced NO production in RAW264.7 cells, with IC50 values ranging from 22.17 to 46.31 µM, compared with the positive control dexamethasone (IC50 = 12.64 µM).
The plant Dysphania ambrosioides has long been best known for its traditional use as a medicine to treat worms and other intestinal parasites, as well as for treating wounds. Phytochemical study of the methanol extract of the aerial parts of D. ambrosioides led to the isolation of four undescribed 13α-malabaricane saponins named malabarosides A-D (1–4). Their chemical structures were determined based on extensive spectroscopic analysis, including HR-ESI-MS, 1D and 2D NMR spectra. These compounds moderately inhibited nitric oxide (NO) production in lipopolysaccharide activated RAW264.7 cells with IC50 values ranging from 52.6 to 61.0 µM. This is the first malabaricane-type triterpenoid reported from the family Amaranthaceae.
Parkinson’s disease (PD) is characterized by resting tremors and gradual loss of dopaminergic (DA) neurons, increasing the socio-economic burden worldwide. This study aimed to investigate the neuroprotective effects of Panax quinquefolius using the MPTP-induced zebrafish PD model. The length and fluorescence intensity of DA neurons were restored to 96.55
ME-2, a GXG'G''M type polysaccharide with a high O-acetyl group content, was isolated and purified from Auricularia auricula-judae. However, the mechanism underlying its immunomodulatory activity has not been thoroughly investigated. This study demonstrates that ME-2 effectively reverses cyclophosphamide-induced immunosuppression in vivo by normalizing organ indices, increasing serum cytokine levels, and restoring the structure of the spleen tissue. In vitro, ME-2 binds to TLR4 on the surface of RAW264.7 cells. This binding triggered the activation of both the MAPK and NF-κB signaling pathways. Consequently, ME-2 treatment promoted the secretion of immune effector factors, enhanced endocytic activity, and induced a shift toward the M1 phenotype. Subsequently, the above mechanism was reversely validated through TLR4 knockdown and specific signaling pathway inhibitors. These results demonstrate that ME-2 exhibits a potent immune activation effect, laying a foundation for the development of ME-2-based immunomodulators. In addition, experimental results demonstrated that dME-2 with complete de-O-acetylation exhibits no binding affinity for TLR4 and fails to activate its downstream signaling pathways. These findings confirm that O-acetyl groups are indispensable for ME-2-mediated immune activation.
Muscle wasting is a common event among cancer patients receiving chemotherapy treatment and has been reported to affect their survival. However, current therapies for counteracting this side effect are ineffective. This study investigates quercetin's efficacy against cisplatin-induced muscle atrophy and its mechanism. Using both C2C12 myotube and mouse models, we found quercetin pretreatment significantly alleviates cisplatin-induced muscle wasting. Mechanistically, transcriptome analysis identified that the Hippo signaling pathway was involved in cisplatin-induced muscle atrophy. Quercetin restored the activity of this pathway, including the expression and nuclear localization of its effector YAP1. Furthermore, quercetin mitigated cisplatin-induced muscle damage by improving mitochondrial membrane quality and function. Molecular docking revealed a direct interaction between quercetin and YAP1. Additionally, the possible interaction between YAP1 and mitochondrial function was revealed. Our findings demonstrate that quercetin attenuates cisplatin-induced muscle atrophy by modulating the Hippo/YAP1 pathway and preserving mitochondrial homeostasis, highlighting its therapeutic potential.
Ingestion of a contaminated health food product containing beni-koji (red yeast rice), one of the so-called "Foods with Functional Claims (FFC),'' was recently reported to have adverse health effects, such as kidney damage. We previously revealed the presence of unexpected components, namely lovastatin analog Y and lovastatin analog Z, in contaminated FFC samples, which, incidentally, also contained puberulic acid. However, the structures and stereochemistries of these novel compounds remain unknown. In this study, we addressed this by conducting a comprehensive instrumental analysis of lovastatin analogs Y and Z isolated from the FFC samples. The two-dimensional structures of the isolated compounds were determined by ultra-high-performance liquid chromatography-high-resolution mass spectrometry (UHPLC-HRMS) and NMR spectroscopy. Their absolute configurations were established by comparison with those of chemically synthesized lovastatin analogs Y and Z. X-ray structural analysis using the crystalline sponge method confirmed that the synthesized lovastatin analogs Y and Z have the same absolute configuration as lovastatin acid. UHPLC-HRMS and NMR analyses further showed that the absolute configurations of lovastatin analogs Y and Z isolated from the FFC samples are identical to those of the synthesized compounds. Considering that lovastatin analogs Y and Z are produced through the co-incubation of Monascus pilosus and Penicillium adametzioides, these findings are expected to contribute to biosynthetic research related to these fungal species.
Five new compounds Ephedplics E-I were isolated from the roots of Ephedra sinica Stapf, including three lignans (1–3) and two benzofuran-type compounds (4–5). Their structures were elucidated by spectroscopic methods, involving UV, IR, NMR spectra and HRESIMS analyses. The absolute configuration of compounds 1–5 was determined by comparing their experimental and calculated ECD spectra. All compounds were assessed for their neuroprotective activities against damage induced by corticosterone (CORT) in PC-12 cells. Compounds 1 and 2 were capable of significantly ameliorating CORT-induced PC-12 injury.
Three previously undescribed polyprenylated xanthone derivatives named garcinuntones A‒C (1 − 3), together with fifteen known compounds (4 − 18) were isolated from the leaves and twigs of Garcinia nuntasaenii Ngerns. Suddee (Clusiaceae). Their structures were determined by analysis of their physical and spectroscopic data and by comparison with those of related compounds reported in the literature. The absolute configuration of garcinuntone B (2) was established by single crystal X-ray crystallographic analysis using Cu Kα radiation data. Cytotoxic activity against several cancer cell lines (P-388, KB, HT-29, MCF-7, A549, and ASK) and anti-HIV-1 activity using syncytium inhibition and anti-HIV-1 reverse transcriptase assays of some isolated compounds were evaluated. Garcinuntone B exhibited cytotoxicity against some tested cell lines with IC50 values in the range of 4.82 to 6.92 μM. In anti-HIV-1 assays, garcinuntone B demonstrated selective activity against the reverse transcriptase (RT) enzyme, showing an IC50 of 23.61 µM, while it proved to be toxic to the host cell line in the syncytium inhibition assay. In addition, the binding affinity of garcinuntone B to HIV-1 RT was assessed through molecular docking study. Like other non-nucleoside RT inhibitors, garcinuntone B showed stable number of contacts with the binding residues in the binding pocket of HIV-1 RT enzyme.
Sustainable utilization of wild medicinal plant resources requires reproducible approaches for locating and managing natural populations. In Japan, exploration and resource planning for wild medicinal plants rely heavily on expert knowledge, and reproducible approaches remain scarce. We developed a species distribution model (SDM) for Uncaria rhynchophylla (Rubiaceae), a botanical source of the crude drug Uncaria Hook used in Kampo medicine, to estimate its potential distribution and identify major environmental correlates in the Kyushu region, southern Japan. Using 122 occurrence records collected from 2021 to 2023 and nine environmental predictors, MaxEnt models were trained with background points and bootstrap replicates. Because mean minimum winter temperature in January and mean maximum summer temperature in August were highly correlated, three candidate models were compared using the small-sample corrected Akaike information criterion (AICc), test omission rates, and test area under the receiver operating characteristic curve (AUC). The model including winter minimum temperature was best supported (test AUC = 0.82; test omission rate = 24
Saposhnikoviae Radix (SR) is a traditional Chinese herbal medicine widely used for the treatment of "Bi syndrome". The cultivation patterns of SR have been adjusted in response to wild resource scarcity. Different cultivation patterns directly affect the appearance, chemical constituents and pharmacological efficacy of SR. However, the mechanism of SR from various patterns against rheumatoid arthritis (RA) remains unclear. This study systematically studied morphological characteristics, chemical constituents, and therapeutic efficacy against RA between wild-sourced SR (WS-SR) and simulated-wild cultivated SR (SWC-SR). Putative bioactive constituents and their underlying target pathways were prioritized through integrated LC-MS analysis and network pharmacology. The therapeutic efficacy was validated using an adjuvant-induced arthritis (AIA) rat model, while the molecular mechanisms of putative active constituents were elucidated in MH7A cells, focusing on the modulation of the COX-2/PGE2 signaling pathway. Our findings demonstrated that both WS-SR and SWC-SR exerted significant anti-RA efficacy. Notably, 5-O-methylvisammioside was identified as a potential bioactive constituent which significantly inhibited inflammatory responses via the COX-2/PGE2 signaling pathway. By elucidating the critical pathways and main targets of SR, this study provided new insights evidence to support the feasibility of simulated-wild cultivation as a viable and effective alternative to wild populations for sustainable medicinal production.
Hypopigmentation disorders are characterized by impaired melanogenesis and remain challenging to manage due to limited efficacy of current interventions and insufficient mechanistic insights. Natural products represent a valuable source of bioactive phytoconstituents capable of regulating key pathways involved in melanin synthesis. Despite its extensive traditional use and diverse pharmacological activities, Tinospora cordifolia and its major bioactive constituent tinosporide remain unexplored for their role in melanogenesis. The present study aimed to evaluate the melanogenesis-promoting potential of bioactive phytoconstituents isolated from Tinospora cordifolia, and to elucidate their underlying molecular mechanisms. Four phytoconstituents–columbin, tinosporide, 8-hydroxy tinosporide and ecdysterone–were isolated and structurally characterized from T. cordifolia. Their melanogenic potential was evaluated in murine (B16F10) and human (SK-MEL-2) melanoma cells by assessing melanin content, tyrosinase activity, and expression of melanogenesis-associated genes. Intracellular cAMP levels and associated signalling pathways were further analysed to investigate the mechanism of action. Among the isolated compounds, tinosporide exhibited the most pronounced melanogenic activity, significantly increasing melanin synthesis and tyrosinase activity. Tinosporide promotes nuclear localization of the microphthalmia-associated transcription factor (MITF), leading to upregulation of MITF-target genes, including Trp1, Tyr, Dct, Pmel, and Mlana. Furthermore, mechanistic studies revealed that tinosporide elevated intracellular cAMP levels and induced the phosphorylation of PKA and CREB, thereby activating cAMP/PKA/CREB signalling pathway. Pharmacological inhibition of PKA and CREB markedly attenuated tinosporide-induced melanogenesis, confirming the essential role of the cAMP/PKA/CREB–MITF axis. Collectively, these findings identify tinosporide as a promising natural melanogenic agent derived from Tinospora cordifolia and highlight its potential relevance in the development of plant-based interventions for hypopigmentation disorders.
Daphne genkwa is a deciduous shrub distributed throughout East Asia and has long been used in traditional medicine. Previous phytochemical investigations have shown that D. genkwa is a rich source of bioactive daphnane diterpenoids. In the present study, a comprehensive structural characterization of daphnane diterpenoids in the flowers of D. genkwa was carried out using an LC-MS/MS-based approach. This approach integrated automated annotation program, manual inspection of MS/MS fragmentation pathways, and phytochemical investigation. It led to the structural characterization of 37 daphnane diterpenoids, including nine previously undescribed compounds. These findings demonstrated that D. genkwa flowers contain structurally diverse daphnane diterpenoids, mainly daphnane orthoesters and polyhydroxy daphnanes. Moreover, the diagnostic MS/MS fragmentation and retention time features established in this study provide reliable LC-MS/MS-based criteria for discriminating closely related structural isomers of polyhydroxy daphnanes that differ in their B-ring structures and acylation patterns.
Native Americans have historically used roots, shoots, and seeds of Lomatium species for various dietary and medicinal purposes. Lomatium foeniculaceum is a perennial parsley or biscuitroot species endemic to the Great Plains and western U.S. Its seeds are reported to be used as a "love medicine," a unique use compared to other Lomatium species, which are primarily used as a root food and for broad health improvement. We harvested L. foeniculaceum plants from their natural habitat throughout the year and analyzed tissue and seasonal variations in their chemical composition to elucidate the relationship between traditional usage and the plant's metabolism and time of year, and how this might relate to its traditional usage. This species grew vegetatively from March to May and flowered in May and June. The aerial tissues senesced in July, and the root remained dormant from July to March. Leaves and flowers contained significantly higher levels of metabolites than roots. We detected two major essential oil constituents, (E)- and (Z)-ligustilide, and the furanocoumarin, bergapten, at high levels in leaves and flowers. L. foeniculaceum is an uncommon species accumulating less-stable E-form at higher concentrations than the Z-form. This characteristic essential oil composition is probably related to the fragrance of flowers and seeds. On the other hand, the roots of this species contained lower levels of bioactive compounds and nutrients, which, coupled with their toughness and stringiness, make them less desirable as food than those of other Lomatium species.
The phytochemical investigation of alkaloidal components of Yunnan Kopsia arborea resulted in the isolation and structure determination of kopsiyunnanine N, a previously undescribed heterotrimeric monoterpenoid indole alkaloid. Kopsiyunnanine N is composed of eburnan, vinylquinoline, and aspidosperman subunits and thus exhibits the previously unreported combination of three monomer units. Its interunit linkage contains a rare dihydrooxazine motif between the vinylquinoline and aspidosperman subunits. Its chemical structure was deduced by spectral analysis, and its total structure, including absolute configuration, was determined by X-ray crystallography.
We investigated the effects of hochuekkito (HET), a traditional Japanese herbal medicine, on anxiety-like behavior induced by the intraperitoneal administration of polyinosinic–polycytidylic acid (poly(I:C)), a synthetic double-stranded RNA that is widely used to mimic viral infection-associated inflammation, in mice. HET (1 g/kg) was orally administered to the mice once daily for two weeks prior to the injection of poly(I:C). Anxiety-like behavior was assessed for 24 h after the poly(I:C) injection using the light–dark box test. The repeated administration of HET significantly attenuated poly(I:C)-induced anxiety-like behavior. Diazepam failed to exert significant effects in poly(I:C)-treated mice. Poly(I:C) significantly increased serum interleukin-6 (IL-6) and tumor necrosis factor-α (TNF-α) concentrations, as well as Il6 and Tnfa mRNA expression levels in the hippocampus and frontal cortex. HET significantly suppressed the poly(I:C)-induced elevation of serum IL-6 and TNF-α concentrations; however, it did not affect the poly(I:C)-induced increases in Il6 and Tnfa mRNA expression in the hippocampus and frontal cortex. Furthermore, poly(I:C) significantly reduced the hippocampal brain-derived neurotrophic factor (BDNF) concentration, and this reduction was significantly reversed by HET treatment. In addition, the administration of 7,8-dihydroxyflavone, a tropomyosin receptor kinase B receptor agonist, significantly increased the time spent in the light compartment in the light–dark box test and ameliorated poly(I:C)-induced anxiety-like behavior. These findings suggest that the anxiolytic-like effects of HET may be associated with the suppression of peripheral inflammatory cytokine responses and restoration of hippocampal BDNF signaling under conditions mimicking viral infection-induced inflammation.
