nsc-106399 and Inflammation

Osteoarthritis is a degenerative joint disease. Cartilage degeneration is the earliest and most important pathological change in osteoarthritis, and persistent inflammation is one of the driving factors of cartilage degeneration. Cucurbitacin E, an isolated compound in the Cucurbitacin family, has been shown to have anti-inflammatory effects, but its role and mechanism in osteoarthritic chondrocytes are unclear.. For in vitro experiments, human chondrocytes were stimulated with IL-1β, and the expression of inflammatory genes was measured by Western blotting and qPCR. The expression of extracellular matrix proteins was evaluated by immunofluorescence staining, Western blotting and saffron staining. Differences in gene expression between cartilage from osteoarthritis patients and normal cartilage were analysed by bioinformatics methods, and the relationship between Cucurbitacin E and its target was analysed by a cellular thermal shift assay, molecular docking analysis and molecular dynamics simulation. For in vivo experiments, knee osteoarthritis was induced by DMM in C57BL/6 mouse knee joints, and the effect of Cucurbitacin E on knee joint degeneration was evaluated.. The in vitro experiments confirmed that Cucurbitacin E effectively inhibited the production of the inflammatory cytokine interleukin-1β(IL-1β) and cyclooxygenase-2 (COX-2) by IL-1β-stimulated chondrocytes and alleviates extracellular matrix degradation. The in vivo experiments demonstrated that Cucurbitacin E had a protective effect on the knee cartilage of C57BL/6 mice with medial meniscal instability in the osteoarthritis model. Mechanistically, bioinformatic analysis of the GSE114007 and GSE117999 datasets showed that the PI3K/AKT pathway was highly activated in osteoarthritis. Immunohistochemical analysis of PI3K/Akt signalling pathway proteins in pathological slices of human cartilage showed that the level of p-PI3K in patients with osteoarthritis was higher than that in the normal group. PI3K/Akt were upregulated in IL-1β-stimulated chondrocytes, and Cucurbitacin E intervention reversed this phenomenon. The cellular thermal shift assay, molecular docking analysis and molecular dynamics experiment showed that Cucurbitacin E had a strong binding affinity for the inhibitory target PI3K. SC79 activated Akt phosphorylation and reversed the effect of Cucurbitacin E on IL-1β-induced chondrocyte degeneration, demonstrating that Cucurbitacin E inhibits IL-1β-induced chondrocyte inflammation and degeneration by inhibiting the PI3K/AKT pathway.. Cucurbitacin E inhibits the activation of the PI3K/AKT pathway, thereby alleviating the progression of OA. In summary, we believe that Cucurbitacin E is a potential drug for the treatment of OA.

Topics: Animals; Chondrocytes; Humans; Inflammation; Interleukin-1beta; Menisci, Tibial; Mice; Mice, Inbred C57BL; Molecular Docking Simulation; NF-kappa B; Osteoarthritis, Knee; Phosphatidylinositol 3-Kinases; Proto-Oncogene Proteins c-akt

Cucurbitacin E (CuE) shows potential to handle airway remodelling. In the current study, the effects of CuE on nicotine-induced airway remodelling were explored by focussing on its interaction with let-7c-5p/NGF axis. The potential microRNA (miR) as the therapeutic target for CuE treatment was determined using a microarray assay. Changes in viability, inflammation and let-7c-5p/NGF pathway in nicotine-treated bronchial epithelial cells (BECs) were detected under CuE treatment (5 μM). The pathways were manipulated with let-7c-5p inhibitor. Mice were subjected to nicotine treatment and handled with CuE. Changes in pulmonary function and structure were detected. Based on the microarray data, let-7c-5p was selected as the therapeutic target. Viability and inflammation of BECs were induced by nicotine and then restored by CuE. At molecular level, nicotine suppressed let-7c-5p while induced NGF, FN1 and COLIA levels. The effects of CuE were counteracted by let-7c-5p inhibition. In a mouse model, nicotine impaired the function and structure of lung, which was attenuated by CuE and then re-impaired by let-7c-5p antagomir. Collectively, CuE protected against nicotine-induced airway remodelling and partially depended on the induction of let-7c-5p; our future work would pay more attention to other downstream effectors of the miR to promote the treatment of nicotine-induced pulmonary disorders.

Topics: Airway Remodeling; Animals; Epithelial Cells; Inflammation; Mice; MicroRNAs; Nerve Growth Factor; Nicotine; Triterpenes

Professional athletes conduct high-intensitive hypoxic training often accompanied by the increase of many inflammatory-related cytokines and immunosuppression. Cucurbitacin E (CucE), as a triterpenoid isolated from Cucurbitaceae plants, exert potential anti-cancer and anti-inflammatory. However, it is unknown whether that the CucE could be used as dietary supplement for athletes to improve inflammatory response and immunosuppression. In this study, we established the simulative hypoxic training rat and monkey models and evaluated the effects of CucE on immune- and inflammation-related factors. Obvious improvement on pro-inflammatory factors and pro-lymphocyte proliferation activities were showed in CucE treated rats compared with the control. Further supplement of CucE in professional meals for cynomolgus monkeys with 4-weeks high-intensitive hypoxic training also exert effects on altitude-induced oxidative stress, inflammation and immunologic function. Furtherly, we explored the underlying mechanism of CucE in human Jurkat T cells and results showed that CucE may exhibit immunosuppressive effect by attenuating critical cytokine expression through down-regulating the NF-κB signaling pathway. In conclusion, CucE is expected to be a potential dietary supplement for athletes to ameliorate the inflammation and immunosuppression caused by high-intensitive exercise.

Topics: Altitude; Animals; Cytokines; Humans; Inflammation; Oxidative Stress; Rats; Triterpenes

Asthma is a chronic inflammatory disorder of airway affecting people from childhood to old age, and is characterized by airway epithelial dysfunction. Cucurbitacin E (CuE), a tetracyclic triterpene isolated from Cucurbitaceae plants, has been recently proved to exert anti-inflammation and immunology regulation activities. Nevertheless, its roles in asthma remains poorly defined. In the current study, CuE had little cytotoxicity on cell viability of human bronchial epithelial cell line BEAS-2B. Moreover, lipopolysaccharide (LPS) exposure inhibited cell viability and induced cell apoptosis, which was reversed following CuE pretreatment. Additionally, CuE administration suppressed LPS-induced inflammatory cytokine production, including TNF-α, IL-6, and IL-8. Simultaneously, supplementation with CuE decreased the transcripts and releases of mucin 5AC (MUC5AC) in LPS-treated BEAS-2B cells. Intriguingly, CuE inhibited LPS-evoked activation of the high-mobility group box1 (HMGB1)-TLR4-NF-κB signaling by reducing the expression of HMGB1, TLR4 and p-p65 NF-κB. Notably, restoring this pathway by elevating HMGB1 expression largely offset the protective function of CuE against LPS-triggered cell injury, inflammatory response and MUC5AC expression. Consequently, these findings highlight that CuE can ameliorate human bronchial epithelial cell insult and inflammation under LPS-simulated asthmatic conditions by blocking the HMGB1-TLR4-NF-κB signaling, thereby supporting its usefulness as a promising therapeutic agent against asthma.

Topics: Apoptosis; Asthma; Bronchi; Cell Line; Cells, Cultured; Epithelial Cells; HMGB1 Protein; Humans; Inflammation; Interleukin-1beta; Lipopolysaccharides; Mucin 5AC; NF-kappa B; Signal Transduction; Toll-Like Receptor 4; Transcription Factor RelA; Triterpenes; Tumor Necrosis Factor-alpha

Emerging evidence suggests that neuroinflammatory responses are involved in the neuronal injury. Neuroinflammatory response is mediated by cellular components such as microglia and molecular components, including nitric oxide, prostaglandins and inflammatory cytokines, activation of complement proteins etc. Cucurbitacins is a class of highly oxidized tetracyclic triterpenoids isolated mainly from Cucurbitaceae but also from other plan families and has been reported to have pharmacological activities. The present study aimed to investigate the anti-neuroinflammatory effects of Cucurbitacins on TLR 2/4 agonists (amyloid-β, LTA, and LPS)-induced neuroinflammatory response in microglia and the underlying mechanism for Nrf2/ARE pathways. Results indicates that pretreatment with Cucurbitacins significantly reduced the pro-inflammatory cytokine (TNF-α, IL-1β and IL-6) and attenuated iNOS and COX-2 expression in TLR 2/4 agonists-stimulated microglia. In addition, Cucurbitacins inhibited JNK and p38 MAPKs activation and attenuated JAK-STAT and NF-κB activation in TLR 2/4 agonists-stimulated microglia. Next, we evaluate the potential involvement of Cucurbitacins in the activation of Nrf2/ARE signaling pathways and phase II detoxification enzymes activity. Results indicate that Cucurbitacins markedly promoted the activation of Nrf-2/ARE pathway-related downstream factors including NQO-1 and HO-1. Furthermore, anti-neuroinflammatory effects of Cucurbitacins are attenuated in the knockdown of Nrf2, HO-1 and NQO-1 respectively. Cucurbitacins also has neuroprotective effect against microglia over-activation related neuronal damage. This study demonstrates that Cucurbitacins is potent activator of the Nrf2/ARE pathway and is therapeutically relevant not only to neuroinflammatory responses of microglia but also neuroinflammation mediated neuronal injury.

Topics: Amyloid beta-Peptides; Animals; Anti-Inflammatory Agents; Antioxidant Response Elements; Cell Survival; Cells, Cultured; Culture Media, Conditioned; Inflammation; Janus Kinases; Lipopolysaccharides; Mice, Inbred ICR; Microglia; Neurons; Neuroprotective Agents; NF-E2-Related Factor 2; NF-kappa B; Signal Transduction; STAT Transcription Factors; Teichoic Acids; Toll-Like Receptor 2; Toll-Like Receptor 4; Triterpenes

The in vivo and in vitro mechanistic anti-inflammatory actions of cucurbitacin E (CE) (Citrullus lanatus var. citroides) were examined. The results showed that LPS/INF-γ increased NO production in RAW264.7 macrophages, whereas L-NAME and CE curtailed it. CE did not reveal any cytotoxicity on RAW264.7 and WRL-68 cells. CE inhibited both COX enzymes with more selectivity toward COX-2. Intraperitoneal injection of CE significantly suppressed carrageenan-induced rat's paw edema. ORAC and FRAP assays showed that CE is not a potent ROS scavenger. It could be concluded that CE is potentially useful in treating inflammation through the inhibition of COX and RNS but not ROS.

Topics: Animals; Anti-Inflammatory Agents; Carrageenan; Cell Line; Citrullus; Cyclooxygenase 2 Inhibitors; Cyclooxygenase Inhibitors; Edema; Humans; Inflammation; Interferon-gamma; Lipopolysaccharides; Mice; NG-Nitroarginine Methyl Ester; Nitric Oxide; Phytotherapy; Plant Extracts; Reactive Nitrogen Species; Triterpenes