Eight previously undescribed protolimonoids, including seven apotirucallane-type (1, 3, 6, and 14-17) and one tirucallane-type (11) triterpenoids, along with ten known analogues, were isolated from the root bark of Dictamnus dasycarpus Turcz. Their structures were unambiguously determined by comprehensive spectroscopic analysis and single-crystal X-ray diffraction. The absolute configurations of the flexible tetrahydrofuran side chains in 1 and 10 were definitively established, resolving a longstanding stereochemical challenge in this class of triterpenoids. All isolated compounds were evaluated for hepatoprotective, antifibrotic, and cardioprotective activities. Compounds 2, 3, 7, 8, 12-14, and 16-18 exhibited striking hepatoprotective activity against acetaminophen-induced toxicity in HepG2 cells. Compounds 4, 5, 11-13, and 15-17 showed significant inhibitory activity against transforming growth factor-β-induced fibrosis in HSC-T6 cells. Additionally, compounds 1-3, 7, 8, 10, and 11 demonstrated protective effects against hypoxia/reoxygenation-induced injury in H9c2 cells. These findings highlight the therapeutic potential of protolimonoids from D. dasycarpus and provide valuable structure-activity relationship insights for future drug development.

Five undescribed neolignans (1-5), and three known compounds (6-8) were isolated from the root bark of Magnolia officinalis Rehd. et Wils. Compounds 3 and 4 were obtained as racemates and separated completely using a chiral column. Their structures were elucidated through extensive analyses of 1D and 2D NMR, HRESIMS, and electronic circular dichroism (ECD) calculations. The hepatoprotective activities of all isolates were assessed. Results indicated that compounds 1 and 8 exhibited significant hepatoprotective activity in both APAP- and H2O2-induced liver injury models. Further in vitro experiments demonstrated that compound 1 attenuated apoptosis by reducing reactive oxygen species production and modulating the expression of key apoptotic proteins.

Three new abietane-type diterpenes, named pinusins A-C (1-3) were isolated from Pini Lignum Nodi (the dried tuberculate or branched nodes of Pinus massoniana Lamb.), together with five known diterpenoids (4-8), a flavonoid (9) and a monoterpene (10). The structures of these compounds were elucidated based on detailed spectroscopic analyses (1D, 2D NMR, HRESIMS, IR, and UV), while ECD calculations determined their absolute configurations. All compounds were assayed in vitro for their protective activities against acetaminophen (APAP)-induced hepatotoxicity at 10 μM. Compounds 1-4, 9, and 10 experimentally exerted protective effects against APAP-induced HepG2 cell damage.

Three new diterpenes, named pinusins D-F (1-3), were isolated from the Chinese medicine "Pini Lignum Nodi" (the dried tuberculate or branched nodes of Pinus massoniana), along with twelve known diterpenes. The structures of these new compounds were determined using detailed spectroscopic methods and ECD calculations. Compounds 4-15 displayed moderate hepatoprotective activity against H2O2-induced oxidative damage in HepG2 cells, while compounds 5-15 exhibited markedly enhanced efficacy -surpassing that of bicyclol (positive control).

Hypertension and metabolic dysfunction-associated fatty liver disease (MAFLD) are both common chronic diseases globally. Nearly half of patients with hypertension are complicated by MAFLD. The mechanisms of the bidirectional promotion between the two remain unclear. The (pro) renin receptor ((P)RR) is one of the classic members of the renin–angiotensin system (RAS) and serves as the receptor for prorenin. Although the role of (P)RR in the induction and progression of hypertension has been extensively studied, its role and underlying mechanisms in MAFLD remain underreported. In this study, we aim to investigate the role of (P)RR in the pathogenesis of hypertension combined with MAFLD. In this study, SHRs were used for the model for hypertension combined with MAFLD. Liver lipid content analysis, liver H&E staining, the detection of (P)RR, ERK and downstream proteins related to fatty acid synthesis and transport, and RNA sequencing and data analysis were performed. In the in vitro experiments, we activated (P)RR using renin and established the lipid deposition model of HepG2 cells induced by renin for the first time. (P)RR was specifically blocked using handle region peptide (HRP), and Nile red fluorescence staining, (P)RR/ERK/PPARγ protein expression analysis, and immunofluorescence were performed to further verify the role of (P)RR in the pathogenesis of hypertension combined with MAFLD. Our results demonstrate that (P)RR plays a role in the development and progression of hypertension combined with MAFLD. The hepatic TG and FFA levels in the SHRs were increased, and the protein expression of the (P)RR/ERK/PPARγ pathway and downstream proteins related to fatty acid synthesis and transport were upregulated. HRP reversed the activation of these proteins and reduced intracellular lipid accumulation. In conclusion, our study first reveals that (P)RR is a potential therapeutic target for hypertension combined with MAFLD. And we found the (P)RR/ERK/PPARγ axis for the first time, which plays an important role in the progression of spontaneous hypertension combined with MAFLD.

Polygonati Rhizoma serve as medicinal and edible plants rich in carbohydrates, with its polysaccharides extensively studied, while research on oligosaccharides remains limited regarding their structural and bioactive properties. This study investigates the against non-alcoholic fatty liver disease (NAFLD) effects of Polygonatum kingianum oligosaccharides (PKO) and employs UPLC-Orbitrap-MS/MS technology to characterize oligosaccharide components. The PKO components were enriched through hot water extraction, 60% anhydrous ethanol precipitation, and membrane separation technology, with preliminary validation of their hepatoprotective effects in DL-ethionine-induced fatty liver mice. A novel LC-MS/MS analytical method was developed utilizing characteristic secondary fragment ions and retention time profiles of oligosaccharides to enable rapid identification and characterization of PKO. Thirty-four oligosaccharides were successfully identified from Polygonatum kingianum (PK), all exhibiting a monosaccharide composition of fructose and glucose. The structural backbone consists of fructose residues linked via (1→2) and (2→6) glycosidic bonds, with 32 compounds being reported for the first time in PK. This study marks the first discovery of oligosaccharides in PK with potential therapeutic effects against NAFLD. Additionally, it represents the first application of LC-MS/MS technology to systematically characterize neutral oligosaccharides in this species. These findings provide a novel analytical methodology for investigating potential anti-NAFLD oligosaccharide components in PK.

BACKGROUND:The prevalence of diabetes mellitus-associated metabolic dysfunction-associated fatty liver disease (MAFLD) is on the rise, with over 90% of patients with Type 2 Diabetes Mellitus (T2DM) also suffering from MAFLD. Currently, there are no specific drugs available for the treatment of diabetes mellitus-associated MAFLD. Ginsenoside is commonly used to treat both MAFLD and diabetes. Previous studies have used biosynthesis to obtain the ginsenoside precursor compound 20S-O-Glc-DM (C20DM), which is suitable for industrial production, has high bioavailability, and is promising as a new treatment option for diabetes mellitus-associated MAFLD. PURPOSE:This study is the first to reveal the protective effects of C20DM on diabetes mellitus-associated MAFLD and its potential pharmacological mechanisms. METHODS:We used DB/DB mice fed a high-fat diet as an animal model of diabetes mellitus-associated MAFLD and HepG2 cells treated with high glucose medium combined with oleic acid as an in vitro model. In vivo, eukaryotic transcriptome sequencing and 16S rRNA analysis were used to explore the mechanism of action of C20DM. In vitro, further validation of the relationship between PGC-1α and C20DM in improving diabetes mellitus-associated MAFLD was conducted using the SR18292 inhibitor. RESULTS:In both the in vitro and in vivo models of diabetes mellitus-associated MAFLD, C20DM improved hepatic lipid accumulation and mitochondrial function. 16S rRNA analysis revealed that C20DM ameliorated gut microbiota dysbiosis and increased the abundance of microbiota associated with mitochondrial function. Eukaryotic transcriptome analysis showed that differentially expressed genes were primarily involved in hepatic fatty acid metabolism and the PPAR pathway, with PGC-1α playing a key role. CONCLUSIONS:In conclusion, our study demonstrates that C20DM improves diabetes mellitus-associated MAFLD by regulating fatty acid metabolism and mitochondrial function through the PGC-1α/PPARα/CPT1A pathway.

Atractylodimers A-D (1-4), sesquiterpenoid dimers (SDs) featuring a unique cage-like structure, were isolated from the rhizomes of Atractylodes macrocephala. The most distinctive characteristic of these isolates was the highly twisted "cap" structure based on highly twisted five-membered oxygen heterocyclic rings. Notably, compound 1 contained a furo[2,3b]furan ring, a caged 3,10-oxa-tricyclo[5.2.1.04,9]decane moiety, and 6/6/5/5/5/5/6/6 octocyclic skeleton. Compounds 2-3 exhibited a spirotetrahydrofuran ring, while compound 4 incorporated a caged spiro2,5,9-oxa-tricyclo[5.2.1.04,10]decane scaffold for unit linkage. Their structures were definitively established through spectroscopic methods and X-ray diffraction experiments. Plausible biosynthetic pathways of compounds 1-4 were proposed. Compounds 1 and 2 demonstrated significant neuroprotective effects against serum deprivation-induced PC12 cell damage.

Promiscuous enzymes are distributed widely in nature. These promiscuous enzymes recognize multiple substrates and yield a variety of metabolites. Hence, the enzyme promiscuity is usually avoided to improve the specificity of metabolites. In this study, the enzyme promiscuity, however, is used to diversify flavonol 3-O-galactoside 6''-O-acetates (F3Gal-6''As), which are active compounds in plants. Specifically, a pathway harbouring four promiscuous flavanone 3-O-hydroxylase (CsF3Ha), flavonol synthase (AcFLS-HRB), flavonol 3-O-galactosyltransferase (PhUGT), and acetyltransferase (GAT), was successfully constructed and introduced into Escherichia coli BL21 (DE3) cells to form engineered strains via modular engineering. An engineered strain, MT4, was selected for further optimization. Protein engineering and condition optimization were thus performed to generate a MT4-derived strain MT4-D17W capable of synthesizing multiple F3Gal-6''As. The medium-dependent yields of three F3Gal-6''As, trifolin 6''-O-acetate (T6''A), hyperoside 6''-O-acetate (H6''A), and myricetin-3-O-(6''-O-acetyl)-galactoside (MGal6''A), in the shake-flask culture of MT4-D17W were 404.4, 255.5, and 117.1 mg/L, respectively. The T6''A yield in the 6-L reactor reached 750.8 mg/L. These enzymatically synthesized F3Gal-6''As displayed an increased liposolubility, while T6″A and MGal6″A displayed hepatoprotective activity. This research not only provides a new insight for the promiscuous enzymes, but also lays a foundation for the scale preparation of flavonol 3-O-galactoside 6''-O-acetates.

Six sulfur-substituted triptolide (TPL) analogs (STP1-6) were synthesized and evaluated for their biological functions. Among them, STP2 had significant antitumor activity both in vitro and in vivo. Notably, the intraperitoneal injections of 1 g/kg STP2 did not cause mice death and apparent pathological damage, while the mice in the TPL group (2 mg/kg) lost weight and all died within 4 days. The antitumor effect of STP2 could mediated by the inhibition of SRSF1 expression to regulate Bcl-x pre-mRNA splicing, which in turn induces autophagy and promotes cell death. This mechanism was the first time discovered in the field of TPL research. These results indicated that compound STP2 could be a promising lead compound for further studies.

Stiff skin syndrome is a very rare non-inflammatory reactive skin disease, characterized by skin sclerosis and limited joint mobility. The paper reports one case of child with stiff skin syndrome and treated with combined therapy of acupuncture and cupping. Acupuncture was used at the lateral line 1 of vertex (MS8) on the right side, Jiaji (EX-B2) of L2 to L4, Huantiao (GB30), ashi point, Juliao (GB29), Fengshi (GB31), Weizhong (BL40), and etc. on the left side. After deqi, the electrodes of KWD-808Ⅰimpulse electronic therapeutic device were attached to Jiaji (EX-B2) of L4 and Huantiao (GB30), Fengshi (GB31) and Yanglingquan (GB34) on the left side respectively, at disperse-dense wave, a frequency of 2 Hz/100 Hz, and a current of 2 mA. The needles were retained for 20 min. Acupuncture was operated once every 2 days, 3 interventions a week. When acupuncture was completed in each intervention, moving cupping was followed till the skin turned to be red, along the distribution of the governor vessel, foot-shaoyang gallbladder meridian and foot-taiyang bladder meridian on the left side, of the lumbar region and leg. Moving cupping was delivered once every 2 days, 3 times a week. Once a week, after moving cupping, the cups were retained on the areas with skin stiffness for 8 min to 10 min. One course of the combined therapy of acupuncture and cupping was composed of 6 treatments. After 2 courses of treatment, the skin stiffness on the left buttock region and the lateral side of the lower limb was ameliorated, the swelling on the left lower limb relieved and the walking improved; and the patient could walk continuously for 2 000 m. The combined therapy of acupuncture and cupping provides a new idea for the clinical treatment of stiff skin syndrome.

Background Left ventricular diastolic dysfunction (LVDD) is a manifestation of heart failure, with both its incidence and prevalence increasing annually. Currently, no pharmacological treatments are available for LVDD, highlighting the urgent need for new therapeutic discoveries. Ginsenosides are commonly used in cardiovascular therapy. Previous research has synthesized the ginsenoside precursor molecule, 20S-O-Glc-DM (C20DM), through biosynthesis. C20DM shows greater bioavailability, eco-friendliness, and cost-effectiveness compared to traditional ginsenosides, positioning it as a promising option for treating LVDD. Purpose This study firstly documents the therapeutic activity of C20DM against LVDD and unveils its potential mechanisms of action. It provides a pharmacological basis for C20DM as a new cardiovascular therapeutic agent. Methods In this study, models of LVDD in mice and ISO-induced H9C2 cell damage were developed. Cell viability, ROS and Ca2+ levels, mitochondrial membrane potential, and proteins associated with mitochondrial biogenesis and autophagy were evaluated in the in vitro experiments. Animal experiments involved administering medication for 3 weeks to validate the therapeutic effects of C20DM and its impact on mitochondria and autophagy. Results Research has shown that C20DM is more effective than Metoprolol in treating LVDD, significantly lowering the E/A ratio, e'/a' ratio, and IVRT, and ameliorating myocardial inflammation and fibrosis. C20DM influences the activity of PGC-1α, downregulates PINK1 and Parkin, thereby enhancing mitochondrial quality control, and restoring mitochondrial oxidative respiration and membrane potential. Furthermore, C20DM reduces excessive autophagy in cardiomyocytes via the AMPK-mTOR-ULK1 pathway, diminishing cardiomyocyte hypertrophy and damage. Conclusions Overall, our research indicates that C20DM has the potential to enhance LVDD through the regulation of mitochondrial quality control and cellular autophagy, making it a promising option for heart failure therapy.

Cisplatin is a widely used drug for the clinical treatment of tumors. However, nephrotoxicity limits its widespread use. A series of compounds including eight analogs (G3-G10) and 40 simplifiers (G11-G50) were synthesized based on the total synthesis of Psiguamer A and B, which were novel meroterpenoids with unusual skeletons from the leaves of Psidium guajava. Among these compounds, (d)-G8 showed the strongest protective effect on cisplatin-induced acute kidney injury (AKI) in vitro and vivo, and slightly enhanced the antitumor efficacy of cisplatin. A mechanistic study showed that (d)-G8 promoted the efflux of cisplatin via upregulating the copper transporting efflux proteins ATP7A and ATP7B. It enhanced autophagy through the activation of the adenosine monophosphate-activated protein kinase (AMPK) signaling pathway. (d)-G8 showed no acute toxicity or apparent pathological damage in the healthy mice at a single dose of 1 g/kg. This study provides a promising lead against cisplatin-induced AKI.

We established myocardial injury models in vivo and in vitro to investigate the cardioprotective effect of gomisin D obtained from Schisandra chinensis. Gomisin D significantly inhibited isoproterenol-induced apoptosis and hypertrophy in H9C2 cells. Gomisin D decreased serum BNP, ANP, CK-MB, cTn-T levels and histopathological alterations, and inhibited myocardial hypertrophy in mice. In mechanisms research, gomisin D reversed ISO-induced accumulation of intracellular ROS and Ca2+. Gomisin D further improved mitochondrial energy metabolism disorders by regulating the TCA cycle. These results demonstrated that gomisin D had a significant effect on isoproterenol-induced myocardial injury by inhibiting oxidative stress, calcium overload and improving mitochondrial energy metabolism.

To find structurally previously undescribed compounds with pharmacological effects from Prismatomeris tetrandra (Roxb.) K. Schum (Rubiaceae), thirteen undescribed tetrahydroanthraquinones (1⎼13) named prisconnatanones J⎼V and seven known anthraquinones (14⎼20) were isolated and characterized. The structures of these compounds were elucidated by detailed spectroscopic analyses, and their absolute configurations were established by modified Mosher's method and ECD calculations. The antitumor cell proliferative activities of prisconnatanones J⎼V were determined. Among them, prisconnatanones J possessed high antitumor cell proliferation in HGC27 cells (IC50, 0.792 μM) by blocking HGC27 cells in the S phase and significantly inducing apoptosis in HGC27 cells. Prisconnatanone J has no cytotoxicity to normal gastric cells line (GES-1) at 10 μM and showed a considerable selectivity for HGC27 cells. Prisconnatanone J can potentially inhibit tumor cell proliferation and should be further investigated.

Four undescribed sesquiterpenes, atramacrolodes A-D (1-4), along with six known compounds 5-10 were isolated from the rhizome of Atractylodes macrocephala. Compound 3 possessed a new skeleton based on an unprecedented carton-carton connection. Their structures were determined by UV, IR, HRESIMS, NMR spectra, 13C NMR calculation with DP4+ analysis, and the comparison of experimental and calculated ECD spectra. Compounds 5 and 8 showed protective effects against paracetamol-induced liver cell injury.

Green acetylation of puerarin to form puerarin 6′′-O-acetate using engineered Escherichia coli capable of synthesizing acetyl-CoA.

Magterpenes A–C (1–3), three unprecedented meroterpenoids featuring a unique 6/6/6/6/6 polycyclic skeleton, were isolated from the ethanol extract of Magnolia officinalis Rehd. et Wils. The compounds were obtained as racemic mixtures that were completely resolved through chiral columns. Their structures were elucidated by extensive analyses of one-dimensional (1D) and 2D nuclear magnetic resonance, high-resolution electrospray ionization mass spectrometry, chemical calculations of 1H/13C NMR, and electronic circular dichroism calculations. The compounds were constructed via two Diels–Alder reactions in the proposed biosynthetic pathway. All isolates were evaluated for their nephroprotective and hepatoprotective activities. The results demonstrated that (+)-1 and (−)-1 possessed promising nephroprotective activities in a dose-dependent manner, while (−)-2 and (+)-3 exhibited moderate hepatoprotective activities.