The Study Of The Anti-Inflammatory Activity And Its Mechanisms Of Neuropeptide FF And Novel Chiral Spirooxindole Compound

Inflammation is a complex pathological process that causes many chronic diseases. In the present study, the anti-inflammatory activity of two types of potential therapeutics, neuropeptide FF and chiral spirooxindole compound JP-8g, were investigated to provide novel therapeutic agents for the anti-inflammatory drug development.Neuropeptide FF (FLFQPQRFa) is an octapeptide that isolated from mammals in1985. Previous studies have shown that NPFF is capable of regulating a variety of physiological functions, including cardiovascular, gastrointestinal motility, diet and pain transduction. However, the effect of NPFF in anti-inflammatory field, has not been reported yet. In the present study, we report for the first time that NPFF is capable of modulating inflammatory process. By using reverse transcription polymerase chain reaction (RT-PCR), western blot and immunofluorescence experiments, we observed that NPFF2receptor is naturally functionally expressed in differentiated neuroblastoma cell line dSH-SY5Y, and NPFF activated the NF-κB signal pathway at this cell model. Subsequently, two inflammatory cell models, the primary peritoneal macrophages and RAW264.7macrophages, were used to investigate the anti-inflammatory activity of NPFF in vitro. NPFF2receptor was also found naturally functionally expressed in these two cell models. Moreover, NPFF NPFF dose-dependently modulated the cell proliferation and nitric oxide (NO) production of macrophages. Furthermore, NPFF exerted anti-inflammatory activity in carrageenan-induced foot swelling inflammation mice model in vivo, which may be involved by NPFF receptors. Collectively, these lines of evidence suggest that NPFF is capable of modulating the inflammation process.Non-steroidal anti-inflammatory drugs (NSAIDs) are among the most widely used prescription and nonprescription drugs in the world. However, the side effects of NSAIDs, such as gastrointestinal damage, severally limit the widely clinical application of NSAIDs. In the present study, the anti-inflammatory potency of JP-8g, a novel chiral spirooxindole compound synthesized by our laboratory, was investigated. Our findings including as follows:by using computer molecular docking method and kinase inhibition experiments, JP-8g was found to inhibit the cyclooxygenase (COX) activity in vitro, suggesting that JP-8g belongs to the family of the cyclooxygenase inhibitors; by using primary peritoneal macrophage as cell model, JP-8g was observed to suppress the production of NO in a dose-dependent manner; by using inflammation animal models, JP-8g was found to possess good peripheral (ear edema and foot swelling models in mice) and central (acute neuroinflammation model in mice) anti-inflammatory activity; by using rat stomach injury model, JP-8g was observed to cause very low gastrointestinal toxicity; by using pharmacological antagonists, COX and nitric oxide synthases were found to present the potential targets of JP-8g. Collectively, our data suggest that novel chiral spirooxindole compound JP-8g represents a new type of COX inhibitor with good anti-inflammatory activity and low gastrointestinal toxicity. Furthermore, our present findings not only provide new promising anti-inflammatory therapeutic agents, but also reveal novel mechanism of COX inhibitor, which will be helpful for the development of anti-inflammatory candidates in future.