Assessing The Anti-inflammatory Activity Of Drugs And Their Applications Based On Zebrafish Model

Objective Inflammation is the protective immune response of the innate immune system to harmful stimuli(such as pathogens,dead cells or stimuli),which is strictly regulated by the human body.Excessive inflammation can lead to chronic or systemic inflammation,leading to various inflammation-related diseases.At present,there are a large number of patients suffering from inflammation-related diseases,which has caused a huge economic and disease burden on society and individuals,and has become a major public health problem.The study of new antiinflammatory compounds with good efficacy and safety is a new direction for disease prevention and treatment in the future,and zebrafish as an excellent model organism has excellent prospects in the development of anti-inflammatory compounds.Therefore,we developed a new type of phenanthroimidazole derivative,and studies its anti-inflammatory effect and its mechanism by constructing a zebrafish inflammation model,which provides reference for the in vivo anti-inflammatory treatment and the other application(such as tumors).Methods(1)Transgenic zebrafish Tg(coro1a: e GFP)was taken to cross-cut the tail fin to construct an in vivo inflammation model,and to evaluate the effect of phenidimidazole derivatives on macrophages and neutrophils in the wound;To construct an inflammation model through lipopolysaccharide-induced inflammation in zebrafish,and evaluate the effects of phenanthrmidazole derivatives on apoptotic cells induced by inflammation;(2)By constructing an inflammation model through lipopolysaccharide-induced inflammation in zebrafish,the study was found that the anti-inflammatory effect of phenoimidazole derivatives were exerted by affecting the production of oxidative stress(ROS),nitric oxide(NO)and inflammatory factors(IL-1,IL-6 and TNF-α)in zebrafish.(3)Evaluate the application of phenanthroimidazole derivatives in the field of anti-tumor by in vitro MTT method and in vivo liver cancer zebrafish model;(4)The zebrafish embryos were continuously exposed to phenanthrimidazole derivatives,and the safety of the phenanthrimidazole derivatives was evaluated by counting zebrafish embryo mortality,hatching rate and deformity rate.Results(1)The anti-inflammatory effect evaluation results show that phenanthrimidazole derivatives can inhibit the accumulation of macrophages and neutrophils in zebrafish wounds and promote the elimination and absorption of inflammatory cells,which indicates that the compounds are effective in all periods of inflammation.In addition,phenanthrimidazole derivatives can inhibit the apoptosis induced by inflammation.(2)The results of anti-inflammatory mechanism research found that phenanthrimidazole derivatives can effectively inhibit the production of reactive oxygen species(ROS),NO and inflammatory factors(IL-1,IL-6 and TNF-α)in the zebrafish inflammation model,indicating Phenanthimidazole derivatives can relieve oxidative stress(ROS)and regulate the process of inflammation by inhibiting the release of IL-1,IL-6 and TNF-α and reducing the production of NO.(3)After applying the compound to the field of anti-tumor,it was discovered that the phenanthrimidazole derivative inhibits the proliferation of tumor cells in vivo and in vitro,and exhibits anti-tumor activity.(4)The safety evaluation results indicate that when zebrafish is 96 hpf,zebrafish embryos cannot be hatched in high concentration(≥160μM)of phenidimidazole derivatives,but there is no death or morphological abnormality in each concentration range.As reported in the reference literature,zebrafish died when the concentration of aspirin was 11.1μM.Combined with this study,the acute toxicity of the new phenanthrimidazole derivatives is relatively small.Conclusion Based on the constructed zebrafish inflammation model,the anti-inflammatory activity and mechanism of the new compound can be efficiently evaluated,and it is found that phenanthroimidazole derivatives have a good inhibitory effect on the number of inflammatory neutrophils and macrophages,showing a certain degree Inhibit the apoptotic damage induced by inflammation.Part of the mechanism is to regulate the release of inflammatory factors(IL-1,IL-6 and TNF-α)and reduce the production of NO to relieve oxidative stress(ROS)and regulate the process of inflammation.In addition,through the use of zebrafish,a model organism,it was discovered that phenanthroimidazole derivatives can also be applied to the development and utilization of anti-tumor drugs.At the same time,the developmental toxicity and acute toxicity of phenanthrimidazole derivatives are small,and they have broad application prospects in the future.