Ankyrin repeat domain-containing protein 55 (ANKRD55) is genetically associated with multiple sclerosis, but its cellular function remains incompletely defined. To investigate its potential role in neuroinflammatory pathways, we mapped ANKRD55-interacting proteins in neuroblastoma, astrocytic, microglial, and monocytic cell lines by affinity purification–mass spectrometry. Across all four cell lines, 14-3-3 isoforms were the most reproducible partners, whereas the IMhu-M microglial interactome uniquely contained eight intraflagellar transport complex B (IFT-B) components. In visible immunoprecipitation assays, IFT46–IFT56 was the smallest tested IFT-B subassembly sufficient to support detectable association with ANKRD55. Confocal microscopy localized endogenous ANKRD55 to pericentriolar material and basal bodies, but not to primary cilia, and identified partial association with P-body markers. These data distinguish a conserved 14-3-3 interaction core from cell-restricted centrosomal/IFT-B and RNP-associated ANKRD55 pools.
Obesity is a metabolic syndrome characterized by the excessive accumulation of adipose tissue. Ursolic acid (UA) modulates diverse metabolic processes, including the amelioration of metabolic syndrome and attenuation of hepatic steatosis. However, the specific effects and underlying mechanisms by which UA ameliorates high-fat diet (HFD)-induced obesity and associated metabolic disorders remain to be elucidated. In this study, we investigated the ameliorative effects of UA on obesity using an HFD-induced obese mouse model and a sodium oleate-induced HepG2 cell model. Our results demonstrated that UA supplementation significantly reduced HFD-induced weight gain and visceral fat mass, lowered serum and hepatic total cholesterol (TC), triglyceride (TG), and low-density lipoprotein cholesterol (LDL-C) levels while increasing high-density lipoprotein cholesterol (HDL-C) levels, attenuated hepatic steatosis with a marked reduction in hepatic TG content, and suppressed inflammation with decreased serum and hepatic interleukin-18 (IL-18) and interleukin-1β (IL-1β) levels. Correspondingly, in HepG2 cells, UA mitigated sodium oleate-induced TG and TC accumulation, decreased intracellular ROS, and lowered IL-18 and IL-1β secretion. Additionally, UA upregulated the lipolytic enzymes adipose triglyceride lipase (ATGL) and hormone-sensitive lipase (HSL) at both protein and mRNA levels, downregulated the lipogenic factors sterol regulatory element-binding protein 1c (SREBP1c) and stearoyl-CoA desaturase 1 (SCD-1), and suppressed absent in melanoma 2 (AIM2) and apoptosis-associated speck-like protein containing a CARD (ASC) expression. Notably, we identified growth differentiation factor 15 (GDF15) as a critical upstream mediator of UA’s multifaceted effects. UA robustly upregulated GDF15 expression both in vivo and in vitro, and siRNA-mediated knockdown of GDF15 completely abolished UA’s beneficial actions, including its regulation of lipolysis/lipogenesis, inhibition of the AIM2 inflammasome, reduction in oxidative stress, and attenuation of lipid accumulation. In summary, our findings reveal that UA ameliorates obesity-related metabolic disorders and inflammation by modulating GDF15 expression, which in turn coordinately regulates lipid catabolism/anabolism pathways and AIM2 inflammasome activation. These results establish the UA-GDF15 axis as a promising therapeutic target for the treatment of obesity and its metabolic complications.
Computational intelligence is catalyzing a paradigm shift in protein science, moving from sequence-based homology inference, through template-dependent structure prediction, to data-driven machine learning, and ultimately toward end-to-end foundation models that jointly predict structure and function [...]
Saussurea lappa is a medicinal plant rich in sesquiterpene lactones, particularly costunolide and dehydrocostus lactone. These compounds have attracted interest in their potential applications in cancer and inflammatory diseases. This review examines their molecular actions, critically evaluates the available preclinical evidence, and discusses emerging nanoformulation approaches. Studies suggest that these compounds influence NF-κB, NLRP3, STAT3, PI3K/AKT/mTOR, p53, and MAPK signaling, with associated effects on inflammation, apoptosis, autophagy, and ferroptosis. However, direct molecular targets have been established for only selected mechanisms, and most therapeutic findings remain preclinical. Poor aqueous solubility and limited pharmacokinetic information further complicate their development. Nanoformulation offers a possible approach to improving delivery, with costunolide-loaded bilosomes showing greater cellular activity than the free compound. Nevertheless, evidence of improved systemic exposure and therapeutic efficacy in vivo remains limited. Human studies have mainly examined botanical preparations rather than purified compounds. By bringing together molecular mechanisms, disease-specific evidence, and delivery strategies, this review identifies the main challenges and research priorities for developing costunolide and dehydrocostus lactone as potential therapeutic agents.
Hepatocellular carcinoma (HCC) ranks among the leading causes of cancer death, underscoring the pressing need for therapeutic agents with better efficacy and lower toxicity. Theaflavin (TF) is a polyphenol abundant in black tea that possesses broad-spectrum anti-tumor activity. However, the direct targets and molecular mechanisms of TF in HCC have not been completely defined. Here, reverse virtual screening was employed to predict candidate targets of TF, which were then validated by molecular docking, molecular dynamics (MD) simulation, drug affinity responsive target stability (DARTS), thermal shift assay (TSA) and endogenous fluorescence spectroscopy. The fluorescence titration results showed that TF could directly bind to CDK2, JNK1 and JNK2, with binding free energies of −37.08, −38.77 and −37.27 kJ/mol, and binding constants Ka of 3.16 × 106, 6.25 × 106 and 3.42 × 106 L⋅mol−1, respectively. TF inhibited the viability of HepG2, SK-Hep-1 and Huh-7 cells in a dose-dependent manner, enhanced reactive oxygen species (ROS) accumulation, reduced mitochondrial membrane potential, elevated the Bax/Bcl-2 ratio and activated caspase-9/3, thereby inducing apoptosis through the mitochondrial pathway. Moreover, TF induced G0/G1-phase arrest and curtailed cell migration and invasion, although the latter effects may be partly attributable to reduced cell viability. In nude mice bearing subcutaneous xenografts, TF markedly restrained tumor growth and promoted apoptosis. In summary, TF, a multi-target compound potentially interacting with CDK2 and JNK1/2, was associated with alterations in cell cycle- and MAPK-related signaling and may serve as a potential lead compound for HCC treatment.
