Preliminary Investigation On The Anti-Inflammatory Effect And Mechanism Of Carboxyamido-triazole

OBJECTIVE:Carboxyamido-triazole(CAI) is an experimental anti-cancer agent developed by National Institute of Cancer(NCI) of National Institutes of Health(NIH) of the United States of American.The anti-cancer effects of CAI have been proved by many preclinical experiments and several clinical trails in different stages carried out in the United States.We have found the mechanism of CAI's anti-tumor effect may be associated with inhibition of cell cycle.Meanwhile,we found CAI has a effect of anti-inflammatory.In this paper,we will prove the anti-inflammatory effect of CAI,and research the mechanism in macrophage.Base on these datas we hope to find the same target between anti-tumor and anti-inflammatory effects.Contents:1.Evaluate the effect of CAI on chronic inflammation and immunity inflammation in adjuvant-induced arthritis rat model.2.Use the model of lipopolysaccharide(LPS) stimulated rat alveolar macrophage cell line(NRg383),test the inhibition effect of CAI on TNF-α,IL-1βand NO.And then observe the different in cell signal pathway.Methods:This study adopted adjuvant-induced arthritis rat model to evaluate the effect of CAI on chronic inflammation and immunity inflammation;this study cultured NR8383 cells were stimulated by LPS,added CAI,then use enzyme-linked immunosorbent assay (ELISA) to detect cell supematant TNF-α,NO and IL-1βlevels,and Western blot to detect celt lysates of IκBα,phosphorylated IκBα,and phosphorylated JNK content changes. Results:1.CAI inhibited the primary lesions and secondary lesions of AIA rats at 20 mg/kg,significantly(compare with PEG400 control group,P＜0.05,P＜0.01).2.CAI at 10 and 40μmol·L^-1 both decreased the levels of NO in supematant of NR8383 cell line stimulated by LPS.The level of NO for the DMSO control group was found to be 33.63±0.80μmol·L^-1.Treatment with CAI at 10 and 40μmol·L^-1 decreased the levels to 5.11±0 and 4.43±0.32μmol·L^-1,respectively.The inhibitory rate are 84.8% and 86.8%,respectively.(P＜0.01)3.CAI at 40μmol·L^-1 both decreased the levels of TNF-αin supernatant of NR8383 cell line stimulated by LPS.The level of TNF-αfor the DMSO control group was found to be 224.03±8.94 pg·ml^-1.Treatment with CAI at 40μmol·L^-1 decreased the levels to 185.5±8.53 pg·ml^-1.The inhibitory rate is 17.2%.(P＜0.05)4.CAI at 10 and 40μmol·L^-1 both decreased the levels of IL-1βin supernatant of NR8383 cell line stimulated by LPS.The level of IL-1βfor the DMSO control group was found to be 31.97±3.14 pg·ml^-1.Treatment with CAI at 10 and 40μmol·L^-1 decreased the levels to 13.5±0.9 and 8.43±0.34 pg·ml^-1,respectively.The inhibitory rate are 57.8 and 73.6%,respectively.(P＜0.05,P＜0.01)5.CAI at 10 and 40μmol·L^-1 have no significant effect on IκBα,phosphorylated IκBα, and phosphorylated JNK in which NR8383 cell line stimulated by LPS.Conclusion:CAI can probably play an anti-inflammatory effect via inhibiting cytokines NO,IL-1βand TNF-α,but not acting on the NF-κβand JNK such classical signaling pathways.