An ionic liquid-mediated ultrasonic-assisted extraction (IL-UAE) method was developed for the extraction of Curcuma phaeocaulis polysaccharides (CPPs). The extraction process of IL-UAE was optimized using response surface methodology (RSM) and artificial neural networks (ANN). Compared to the RSM, the ANN model exhibits a higher R2 value (0.9900 vs. 0.9808) and a lower Residual Sum of Squares (26.40 vs. 49.52), indicating that the ANN model has stronger explanatory power and better fitting accuracy. Additionally, this study compared the yield and physicochemical properties of CPPs obtained by the IL-UAE and hot water extraction (HWE). The results indicate that the yield of IL-UAE (3.69 %) was significantly higher than that of HWE (1.51 %). Furthermore, the CPPs extracted using IL-UAE possess higher arabinose and galactose contents, a larger molecular weight, and better anticoagulant activity. Notably, the physicochemical properties and the density functional theory calculations demonstrated that IL-UAE could fragmentate the cell wall and interact with the target polysaccharides. This study offers a promising method for the extraction of functional polysaccharides using environmentally friendly solvents.

Inflammation is a key pathogenic mediator in the development of a multitude of diseases, such as metabolic disorders, cardiovascular events, and neurodegenerative diseases. Inspired by the excellent beneficial effects of Curcuma longa in anti-inflammatory, the EtOH extract of C. longa was phytochemically investigated, which led to the isolation of nineteen sesquiterpenoids (1-19) including eight previously unreported compounds (1-8). Their structures including absolute configurations were established by comprehensive spectroscopic analyses, ECD calculation, and NMR calculation. Furthermore, the anti-inflammatory activities of all the isolates were evaluated in LPS-induced RAW 264.7 cells, and compound 14 exhibited the best anti-inflammatory effect at 50 μM. Mechanistically, RNA sequencing indicated that the anti-inflammatory effect of compound 14 is mediated by the toll-like receptor signaling pathway. Western-blot, CETSA, and DARTS assays exhibited that compound 14 directly targeted TLR4, thereby influencing the TLR4/MyD88/NF-κB signaling pathway to alleviate inflammation. These findings suggested that the sesquiterpenoids from C. longa are a class of potential anti-inflammatory agents.

Atherosclerosis caused by disorders of lipid metabolism is regarded as the main pathological basis of atherosclerotic cardiovascular disease. Inspired by excellent beneficial effects of Ganoderma triterpenoids (GTs) on dyslipidemia, the methanol extract of Ganoderma cochlear was phytochemically investigated, which led to the isolation of 18 GTs including 9 new ones (1 9). Their structures including absolute configurations were established by comprehensive spectroscopic analyses, ECD calculation and NMR calculation. Of these, compound 1 featured a rare 6/4/6/5 tetracyclic system, and compounds 4 and 5 were two unusual 3,4-seco-20,21,22,23,24,25,26,27-octonorlanostane triterpenoids. Furthermore, compound 1 can inhibit lipid accumulation in ox-LDL-induced RAW264.7 cells in a dose-dependent manner. Mechanistically, compound 1 efficiently promoted LXRa nuclear translocation and subsequently accelerated cholesterol efflux by upregulating ABCG1 expression. This investigation illustrated the structural diversity of GTs and revealed that GTs may serve as a class of potential antiatherosclerotic agents.

Background and aims The mechanism of cholestatic liver injury (CLI) is unclear, and effective therapies are lacking. While peroxisome proliferator-activated receptor alpha (PPARα) agonists show potential hepatoprotective effect and pyroptosis is implicated in hepatocellular damage, how PPARα activation mitigates lithocholic acid (LCA)-induced pyroptosis remains unknown. Methods The hepatoprotective effect of PPARα agonists was evaluated in a mouse model of intrahepatic cholestasis induced by LCA. Liver injury was assessed via serum biochemistry, hematoxylin and eosin and TUNEL staining, and electron microscopy. Pyroptosis pathways were analyzed using real-time quantitative polymerase chain reaction, Western blot, and co-immunoprecipitation. Results Combined morphological, histopathological, and biochemical analyses confirmed that PPARα activation protects against CLI. Compared with LCA treatment alone, PPARα activation significantly attenuated the elevation of serum lactate dehydrogenase (LDH), the increased TUNEL-positive cells, and the formation of hepatocyte membrane pores. Mechanistically, PPARα activation suppressed both NOD-like receptor protein 3 (NLRP3) inflammasome-mediated pyroptosis and apoptosis protease-activating factor-1 (APAF-1)/CASPASE-3/GSDME-mediated pyroptosis. Furthermore, PPARα agonist pretreatment inhibited activation of the nuclear factor-kappa B (NF-κB) and forkhead box O1 (FOXO1) signaling pathways. Conclusions PPARα protects against LCA-induced CLI by inhibiting both NLRP3 inflammasome-mediated pyroptosis associated with NF-κB and APAF-1/CASPASE-3/GSDME-mediated pyroptosis associated with the FOXO1 signaling pathway.

Two novel phenolic glycosides, 3-methoxyacetophenone-4-O-(i-D-apiofuranosyl-(1-*2)-(i-D-glucopyranoside (1) and 3-methoxyacetophenone-4-O-(i-D-(5 ''-O-p-hydroxybenzoyl)-apiofuranosyl-(1-*2)-(i-D-glucopyranoside (2), along with one known analogue (3), were isolated from the ethyl acetate extract of Cyathula officinalis. Extensive spectroscopic analyses and chemical methods determined their structures and absolute configurations. The relaxant effects of the isolated compounds on uterine contractions induced by oxytocin were investigated using a rat uterine smooth muscle contraction model. The results indicated that both compounds 1 and 2 had inhibitory effects on oxytocin-induced uterine smooth muscle contraction.

Inspired by the intriguing structures and significant activities of Ganoderma triterpenoids (GTs), the EtOAc extract of Ganoderma applanatum was phytochemically investigated, leading to the isolation of 11 GTs, including 10 new one (1-10). Their structures including absolute configurations, were elucidated through IR, UV, HRESIMS, 1D NMR and 2D NMR data analyses. Notably, applanoids J (1) and K (2) represent the first example of GTs with an unprecedented B-seco-lanostane architecture featuring 1,4-cyclohexanedione motif. Meanwhile, compounds 1-7 and 9-11 were evaluated for their hepatoprotective activity using TGF-β1-induced liver fibrosis model, and some of them such as compounds 2, 4, 6, 7 and 10 significantly suppressed the abnormal upregulation of fibrosis-related genes FN, ACTA2 (encode α-SMA) and COL1A1. Mechanistic studies suggested that the anti-liver fibrosis effect of compound 7 may be mediated through inhibition of the TGF-β/Smad signaling pathway. This study not only illustrates the structural diversity of GTs but also highlights their potential as promising anti-liver fibrosis agents.

Rhododendron, the largest genus of Ericaceae, consists of approximately 1000 species that are widely distributed in Europe, Asia, and North America but mainly exist in Asia. Rhododendron plants have not only good ornamental and economic value but also significant medicinal potential. In China, many Rhododendron plants are used as traditional Chinese medicine or ethnic medicine for the treatment of respiratory diseases, pain, bleeding and inflammation. Rhododendron is known for its abundant metabolites, especially diterpenoids. In the past 13 years, a total of 610 chemical constituents were reported from Rhododendron plants, including 222 diterpenoids, 122 triterpenoids, 103 meroterpenoids, 71 flavonoids and 92 other constituents (lignans, phenylpropanoids, phenolic acids, monoterpenoids, sesquiterpenoids, coumarins, steroids, fatty acids). Moreover, the bioactivities of various extracts and isolates, both in vitro and in vivo, were also investigated. Our review summarized the research progress of Rhododendron regarding traditional uses, phytochemistry and pharmacology in the past 13 years (2010 to December 2022), which will provide new insight for prompting further research on Rhododendron application and drug development.

Pregnane X receptor (PXR) has been considered as a promising therapeutic target for cholestasis due to its crucial regulation in bile acid biosynthesis and metabolism. To search promising natural PXR agonists, the PXR agonistic activities of five traditional Chinese medicines (TCMs) with hepatoprotective efficacy were assayed, and Hypericum japonicum as the most active one was selected for subsequent phytochemical investigation, which led to the isolation of eight nonaromatic acylphloroglucinol-terpenoid adducts including seven new compounds (1 - 4, 5a, 5b and 6). Their structures including absolute configurations were determined by comprehensive spectroscopic, computational and X-ray diffraction analysis. Meanwhile, the PXR agonistic activities of aplenty compounds were evaluated via dual-luciferase reporter assay, RT-qPCR and immunofluorescence. Among them, compounds 1 - 4 showed more potent activity than the positive drug rifampicin. Furthermore, the molecular docking revealed that 1 - 4 were docked well on the PXR ligand binding domain and formed hydrogen bonds with amino acid residues Gln285, Ser247 and His409. This investigation revealed that H. japonicum may serve as a rich source of natural PXR agonists.

Our previous study shows that activation of pregnane X receptor (PXR) exerts hepatoprotection against lithocholic acid (LCA)-induced cholestatic liver injury. In this study we investigated whether PXR activation could inhibit hepatocyte pyroptosis, as well as the underlying mechanisms. Male mice were treated with mouse PXR agonist pregnenolone 16α-carbonitrile (PCN, 50 mg·kg−1·d−1, i.p.) for 7 days, and received LCA (125 mg/kg, i.p., bid) from D4, then sacrificed 12 h after the last LCA injection. We showed that LCA injection resulted in severe cholestatic liver injury characterized by significant increases in gallbladder size, hepatocellular necrosis, and neutrophil infiltration with a mortality rate of 68

(±)-spiroganoapplanin A ( 1 ) with a polycyclic meroterpenoid from Ganoderma applanatum showed potential anti-AD’s effect by reducing Aβ42 production and inhibiting Tau phosphorylation through BACE1, CDK5, and GSK3β-mediated pathways.

Previously, we demonstrated that Schisandrol B (SolB) protected against lithocholic acid (LCA)-induced cholestatic liver injury (CLI) through pregnane X receptor (PXR). Additionally, growing evidence has revealed that pyroptosis is involved in CLI. Whether the hepatoprotective effect of SolB driven by PXR activation is related to pyroptosis in CLI remains unclear. First, the hepatoprotective effect of SolB was confirmed, as evidenced by the decreased mortality, morphological and histopathological changes, and biochemical parameters. The upregulated serum lactic dehydrogenase (LDH) level, increased number of TUNEL-positive cells, and formation of hepatocyte membrane pores induced by LCA were significantly alleviated after SolB pretreatment, indicating that SolB attenuated LCA-induced hepatocyte damage. Further analysis revealed that both NOD-like receptor protein 3 (NLRP3) inflammasome-induced canonical pyroptosis and apoptosis protease activating factor-1 (Apaf-1) pyroptosome-induced noncanonical pyroptosis were significantly inhibited after SolB pretreatment, as illustrated by the decreased expression levels of NLRP3, ASC, caspase-1, and GSDMD and the levels of Apaf-1, caspase-11 p20, caspase-3 p20, and GSDME. Furthermore, the activation of the NF-κB and FoxO1 signaling pathways was inhibited after SolB pretreatment. In addition, the activation of PXR via SolB was proven by luciferase reporter gene assays and the upregulation of PXR targets. The results illustrated that SolB could significantly inhibit NLRP3 inflammasome-induced canonical pyroptosis through the PXR/NF-κB/NLRP3 axis and inhibit Apaf-1 pyroptosome-induced noncanonical pyroptosis through the PXR/FoxO1/Apaf-1 axis. Collectively, this study revealed that SolB protected against CLI by inhibiting pyroptosis through PXR, providing new insights for understanding the molecular mechanism of SolB as a promising anti-cholestatic agent.

Ganoderma triterpenoids(GTs),a class of major active constituents of Ganoderma fungi,possess diverse structures and remarkable activities.In the present study,nine new GTs,namely applanoids A—I(1-9),were isolated from the medicinal fungus of Ganoderma applanatum.Their structures including absolute configurations were established by comprehensive spectroscopic analyses and ECD calculation.Applanoids A—E(1-5)represent the first example of GTs with 6/6/5/6/5 pentacyclic system and the formation of the ether ring between C-15 and C-20 involves Michael addition reaction.Furthermore,compounds 1-8 were evaluated for their hu-man pregnane X receptor(hPXR)agonistic activity using dual-luciferase reporter gene assay,and the results showed that compounds 1,2 and 4 can dose-dependently activate hPXR.This investigation further illustrated the structural diversity of GTs and provided new insights for searching PXR agonists from GTs.

采用硅胶柱色谱、MCI柱色谱、制备薄层色谱和半制备高效液相色谱等分离手段,从黄边灵芝Ganoderma luteomarginatum乙醇提取物的乙酸乙酯萃取部位中分离得到4个羊毛脂烷型三萜化合物,根据NMR、MS、IR数据和X-ray 单晶衍射分析,分别鉴定为(24S,25R)-ganodermanontriol-25-ethyl ether(1)、ganodermanontriol(2)、ganoder-manondiol(3)、hainanaldehyde A(4),其中化合物1为新化合物,所有化合物均为首次从黄边灵芝中分离得到.采用MTT法测定化合物1～3对人肺癌细胞A549、胃癌细胞HGC-27、肝癌细胞SMMC-7721、宫颈癌细胞HeLa的细胞毒活性,结果显示化合物1～3对4种肿瘤细胞具有不同程度的抑制增殖作用,尤其化合物1对A549和HGC-27细胞表现出了显著的细胞毒活性,IC5o分别为4.29±0.89和5.63±0.90μmol·L-1.

Our previous research has shown that lanostane triterpenoids from Ganoderma applanatum exhibit significant anti-adipogenesis effects. In order to obtain more structurally diverse lanostane triterpenoids to establish a structure–activity relationship, we continued the study of lanostane triterpenoids from the fruiting bodies of G. applanatum, and forty highly oxygenated lanostane-type triterpenoinds (1–40), including sixteen new compounds (1–16), were isolated. Their structures were elucidated using NMR spectra, X-ray crystallographic analysis, and Mosher’s method. In addition, some of their parts were evaluated to determine their anti-adipogenesis activities in the 3T3-L1 cell model. The results showed that compounds 16, 22, 28, and 32 exhibited stronger anti-adipogenesis effects than the positive control (LiCl, 20 mM) at the concentration of 20 μM. Compounds 15 and 20 could significantly reduce the lipid accumulation during the differentiation process of 3T3-L1 cells, comparable to the untreated group. Their IC50 values were 6.42 and 5.39 μM, respectively. The combined results of our previous and present studies allow us to establish a structure-activity relationship of lanostane triterpenoids, indicating that the A-seco-23→26 lactone skeleton could play a key role in anti-adipogenesis activity.

Previously, we have demonstrated the antiadipogenic benefits of Ganoderma triterpenoids (GTs), which indicated GTs have potential therapeutic implications for obesity. In this study, the EtOAc extract of Ganoderma applanatum was further phytochemically investigated for searching new antiadipogenic agents, which led to the isolation of a total of 15 highly oxygenated lanostane triterpenoids, including 9 new compounds (1-9) and 6 known analogues (10-15). Structurally, ganodapplanoic acids A and B (1, 2) are two rearranged 6/6/5/6-fused lanostane-type triterpenoids with an unusual C-13/C-15 oxygen bridge moiety. In addition, the EtOAc extract (GAE) and isolates (1-4,6-15) were assayed for their antiadipogenic effects in 3T3-L1 adipocytes. The results revealed that compound 9 effectively repressed adipogenesis through down-regulating the expression of major proteins (PPARγ, CEBPβ and FAS) involving differentiation and adipogenesis in 3T3-L1 adipocytes. Thus, the present study further demonstrated the antiadipogenic potential of GTs and provided a possible perspective for obesity treatment.

Applanmerotic acids A and B (1 and 2) with a polycyclic skeleton isolated from Ganoderma applantum showed anti-inflammatory activity via inhibiting the activation of FPR2.

A pair of new 3',7-epoxy-8,4'-oxyneolignan enantiomers [(+)-1 and (-)-1] as well as a known phenylpropanoid (2) were isolated from the seeds of Croton tiglium Linn. Their structures were established based on extensive spectroscopic analyses. The absolute configurations of (+)-1 and (-)-1 were determined by NMR data calcula-tions and electronic circular dichroism calculations. All compounds were isolated from the genus Croton for the first time. Particularly, (+)-1 and (-)-1 were the first 3',7-epoxy-8,4'-oxyneolignanes reported in Croton. The chemotaxonomic significance of these compounds was discussed.