| Purpose: The model of inflammatory damage were induced by radiation in order to study the mechanism of ferulic acid the to regulate NLRP3 inflammatory bodies at the cell and animal level,and to observe the role of Sirt1 in this regulatory pathway,and to investigate whether ferulic acid could reduce the inflammatory response through Sirt1-NLRP3 inflammatory pathway.This will help to discover radiation protection drugs and molecular mechanisms related to radiation-induced inflammation damage.Methods: The research was divided into three parts.The first part,at the cellular level,the effect of ferulic acid on the inflammatory response of AHH-1 cells induced by radiation were explored,cck-8 method was used to detect the effect of ferulic acid on the activity of AHH-1 cells,and the appropriate dose was determined.Secretion of inflammatory factors were detected by ELISA method,and protein expression in AHH-1 cells after irradiation were investigated by Western Blot method.In the second part,based on the AHH-1 cell inflammation model,the regulation effect of ferulic acid on NLRP3 inflammatory bodies and the mechanism of reducing the inflammation induced by radiation were studied.The effects of ferulic acid on Sirt1 were observed using Sirt1 activators and inhibitors,and the role of ferulic acid in reducing the activation pathway of NLRP3 inflammatory bodies induced by radiation was detected by Western Blot.In the third part,radiation-induced mouse model of immune inflammation damage was established,and in vivo experiments were conducted to further verify that ferulic acid could reduce the inflammatory response caused by radiation through Sirt1-NLRP3 inflammatory pathway.120 male and female C57BL/6 mice were randomly divided into 6 groups:(1)normal control group;(2)radiation model group: 5Gy whole body irradiation;(3)positive drug group,amifostine was intraperitoneal injected at 40 minutes before irradiation;(4)Ferulic acid low dose group: 25mg/kg + 5Gy irradiation;(5)Ferulic acid medium dose group: 50mg/kg + 5Gy whole body irradiation;(6)Ferulic acid high dose group: 100mg/kg + 5Gy whole body irradiation.Mice were administrated with gastric solvent or ferulic acid every day for 12 consecutive days before irradiation.The peripheral blood and serum of mice were collected 24 hours after irradiation.The peripheral blood was used to detect blood leukocytes.The content of inflammatory factors IL-1?,IL-18,IL-6 and TNF-? were observed by ELISA kit.Mouse immune organs spleen and thymus tissues were fixed in 10% formalin solution for pathological testing,immunohistochemistry,and immunofluorescence detection,the other half frozen store in-80 ? refrigerator for Western Blot detection.Results: The results showed that the inflammatory damage model of AHH-1 cells could be established by irradiating at 4Gy,at 12 h after irradiation was the appropriate time to detect the secretion of inflammatory factors and the expression of corresponding proteins.The concentration of ferulic acid(0.01,0.1 and 1 ?mol/L)has good radiation protection effect on AHH-1 cells.According to the above conditions,the regulatory effect of ferulic acid on NLRP3 inflammatory bodies was found at 6h after irradiation.The expression of NLRP3 inflammatory bodies was obvious,and ferulic acid could inhibit the activation of NLRP3 inflammatory bodies in AHH-1 cells induced by radiation.It was further found that ferulic acid could reduce injuries of the radiation-induced AHH-1 cells by increasing the expression of Sirt1 protein,and decreasing the expression of NLRP3 and inflammatory factors.Finally,it was also verified at the animal level that ferulic acid had protective effect on radiation-induced mouse inflammation,significantly reducing the secretion of inflammatory factors in mouse serum and the expression of inflammatory factor proteins in immune organs.Ferulic acid could reverse the reduction of lymphocytes in spleen and thymus tissue of mice induced by radiation.Conclusion: In summary,ferulic acid could reduce the inflammatory effects caused by radiation and could reduce the expression of NLRP3 inflammatory bodies and the secretion of inflammatory factors by activating the expression of Sirt1. |
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