Xiaotansanjiefang, a traditional Chinese medicine formula containing pinellia rhizomes, is investigated for anti-gastric cancer (GC) activity. Cycloartenol, a triterpenoid saponin from pinellia, exhibits emerging anti-tumor properties. This study evaluates its effects on GC cell migration, invasion, and angiogenesis. Cytotoxicity of cycloartenol (0-160 μM) was assessed in GC cells (AGS and MKN-45) and immortalized normal gastric epithelial cells (GES-1) via MTT assays. Colony formation, wound healing, and Transwell assays were conducted to assess proliferation, migration, and invasion. Angiogenesis was evaluated by HUVEC tube formation using conditioned media harvested from cycloartenol-treated GC cells. VEGFA and VEGFR2 protein levels were measured by Western blotting. Cycloartenol showed dose-dependent cytotoxicity in GC cells with minimal toxicity toward GES-1 cells, indicating selectivity. It suppressed colony formation, delayed wound closure, reduced invading cells, and inhibited HUVEC tube formation. Consistently, cycloartenol downregulated VEGFA and VEGFR2 expression in GC cells and HUVECs. In conclusion, cycloartenol selectively suppresses GC cell growth, migration, invasion, and angiogenesis through inactivation of the VEGFA/VEGFR2 pathway.
Allergic rhinitis (AR) is characterized by inflammation of the nasal mucosa caused by an allergic reaction. Cornuside has been reported to possess anti-allergic properties. This study aims to investigate whether cornuside can suppress AR. Mice were induced by ovalbumin (OVA) to establish AR models. The AR symptoms including sneezing and nasal rubbing were recorded, and the eosinophils in mouse mucosal tissues were analyzed by hematoxylin and eosin staining. Total leucocytes in nasal lavage fluid (NLF) were quantified under a hemocytometer. Wright's Giemsa staining was utilized to quantify eosinophils, lymphocytes, and neutrophils in NLF. Enzyme-linked immunosorbent assay was conducted to detect histamine, serum cytokines (OVA-specific IgE, OVAspecific IgG1, interleukin-4, and leukotriene C4), and serum inflammation cytokines (tumor necrosis factor-α, interleukin-6, interleukin-13, and interleukin-17). Cornuside administration significantly reduced sneezing and nasal rubbing behaviors in mice with AR. Mice with AR exhibited more inflammatory cells and higher levels of histamine, serum cytokines, and inflammatory cytokines in the blood or NLF than the control group. However, these upregulations were all reversed by the administration of cornuside. In addition, we found that cornuside treatment improved the inflammatory condition of mice with AR dose-dependently Cornuside can attenuate the inflammatory response in mice with AR.
Rheumatoid arthritis (RA) causes inflammatory bone and cartilage damage. While electroacupuncture (EA) alleviates joint inflammation, its anti-inflammatory mechanisms remain unclear. Sprague-Dawley rats were randomly assigned to control, collagen-induced arthritis (CIA), EA (EA+CIA), and sham-EA (sham-EA+CIA) groups. Rats in the EA group received EA at unilateral ST36 (Zusanli) every other day from day 1. Paw swelling and paw withdrawal latency (PWL) were measured during the treatment. The levels of caspase-1, GSDMD, IL-1β, and IL-18 in the synovium were assessed after 28 days of EA treatment. Our findings demonstrated that inflammatory responses, such as the arthritis index, toe volume, PWL, and inflammatory cell infiltration, were significantly improved in the EA group compared with the CIA and sham-EA groups. Significant decreases in the levels of caspase-1, GSDMD, IL-1β, and IL-18 were observed in the synovium of RA rats that received EA. Furthermore, peripheral blood levels of the inflammatory cytokines IL-1β and IL-18 were significantly reduced in rats in the EA group. Electroacupuncture as implemented at ST36 was associated with reduced caspase-1-mediated pyroptosis-related markers in the joint synovium and decreased IL-1β and IL-18 levels in both synovium and serum in rats with CIA. Acupoint-specificity requires further validation using active-sham or componentresolved control designs.
Osteoarthritis (OA) is a common orthopedic disorder with complicated pathological mechanisms. This study aimed to uncover a novel regulatory network related to vir-like m6A methyltransferase-associated protein KIAA1429 (also known as VIRMA) and ubiquitin-specific protease 3 (USP3) in the progression of OA. An in vitro model of OA was constructed by treating chondrocytes (C28/I2) with interleukin-1β (IL-1β). qRT-PCR and Western blot were applied for gene and protein expression detection. Cell viability was examined using a CCK-8 assay. Cell death was evaluated by flow cytometry following Annexin V/PI staining. Cell senescence was assessed by senescence-associated β-galactosidase (SA-β-gal) staining. Ferroptosis-related events were detected by commercial kits. The interaction between KIAA1429 and USP3 was confirmed via RNA immunoprecipitation assay and dual-luciferase reporter assay. OA mice model was induced by surgical destabilization of the medial meniscus (DMM). Results showed that KIAA1429 expression was up-regulated in C28/I2 cells after IL-1β exposure. Inhibition of KIAA1429 impeded senescence and ferroptosis in IL-1β-treated C28/I2 cells. Mechanistically, KIAA1429 reduced USP3 mRNA expression through m6A methylation modification. Moreover, the protective effects of KIAA1429 knockdown on IL-1β-induced senescence and ferroptosis in C28/I2 cells were rescued by USP3 downregulation. In vivo, KIAA1429 downregulation suppressed the development of OA in DMM mice. These results elucidate that KIAA1429 promotes the senescence and ferroptosis of chondrocytes by increasing m6A methylation of USP3 to suppress its stability, thereby contributing to the pathogenesis of OA.
Non-small cell lung cancer (NSCLC) is a malignant tumor with high morbidity and mortality. Given the reported associations of both SET domain bifurcated histone lysine methyltransferase 1 (SETDB1) with various cancers and ubiquitin-specific protease 7 (USP7) with NSCLC progression, this study aims to elucidate the role of SETDB1 and USP7 in NSCLC progression. mRNA and protein levels were measured by qRT-PCR and Western blot, respectively. Cell biological phenotypes were evaluated using MTT, EdU, flow cytometry, transwell, and sphere formation assays. Ferroptosis was analyzed by measuring Fe2+, MDA GSH, and ROS levels. Relationship between USP7 and SETDB1 was examined through Co-IP, cycloheximide (CHX) chase, and deubiquitination assays. Xenograft tumor models were established to investigate the role of SETDB1 and USP7 in NSCLC in vivo. In NSCLC tissues and cells, SETDB1 levels were elevated. Knockdown of SETDB1 inhibited NSCLC cell viability, proliferation, invasion, and sphere formation ability, while promoting apoptosis and ferroptosis. Mechanistically, USP7 enhanced the stability of SETDB1 protein by its deubiquitination. USP7 knockdown suppressed NSCLC cell biological behavior by downregulating SETDB1. Moreover, USP7 knockdown repressed in vivo tumor growth of NSCLC. Taken together, USP7 suppresses ferroptosis and enhances growth in NSCLC cells by stabilizing SETDB1 protein through deubiquitination.
