Study On The Protection Of Cibai Capsule And Its Effective Components Against Low Dose Radiation

Objective: Explore the protective effects of Cibai capsule and its active components on low dose radiation in mice by establishing a low dose radiation injury model.Investigate the interventional effects of Cibai capsule and its active components on immune and inflammatory related pathways in low dose radiation mice,in order to provide a new therapeutic strategy for low dose radiation protection.Method:C57BL/6J mice were subjected to different low dose irradiation schemes(0.02 Gy,0.04 Gy,0.1 Gy,and 0.2 Gy for 5 consecutive days to accumulate 0.1 Gy,0.2 Gy,0.5Gy,and 1.0 Gy respectively)to cause varying degrees of immune damage.After irradiation,samples were taken and corresponding indicators were measured on days 1,3,7,and 14,respectively.The expression of inflammatory factors in the serum of mice was measured by ELISA,and changes in peripheral blood indicators were measured by blood cell analyzer.Subsets of B lymphocytes and T lymphocytes in the spleen and thymus of mice were analyzed by flow cytometry.The optimal irradiation scheme was selected to establish a low dose radiation injury model in mice,preparing for subsequent drug intervention experiments.C57BL/6J mice were randomly divided into five groups: control group,model group,low dose administration group of Cibai capsule(1 g/kg),medium dose administration group of Cibai capsule(2 g/kg),and high dose administration group of Cibai capsule(4 g/kg).Before irradiation,Cibai capsule was continuously administered by gavage,while the control group and the model group were given physiological saline by gavage once a day for 7 consecutive days.The peripheral blood,spleen,and thymus of mice were collected on the 1st day after irradiation.The total number of leukocytes and lymphocytes in the peripheral blood of mice was measured.The subsets of B lymphocytes and T lymphocytes in the spleen and thymus of mice were analyzed by flow cytometry.The changes of oxidative stress indicators in immune organs and inflammatory factors in serum were detected.The histological changes of spleen in each group were observed by hematoxylin eosin(HE)method.To evaluate whether Cibai capsule has a protective effect on immune damage in mice exposed to low dose radiation,and to predict the potential effective components and potential targets of Cibai capsule against low dose radiation based on network pharmacology.Based on network pharmacology,explore the potential effective components of Cibai capsule to protect against low dose radiation.Through a mouse injury model induced by low dose radiation,give Cibai capsule,syringin,quercetin,kaempferol,and ferulic acid before irradiation to intervene in treatment.After irradiation,the peripheral blood,spleen,and thymus of mice were collected,and the total number of white blood cells and lymphocytes in the peripheral blood of mice was measured.Flow cytometry was used to analyze the subsets of B lymphocytes and T lymphocytes in the spleen and thymus of mice,detect oxidative stress indicators in immune organs,and detect changes in inflammatory factors in serum,in order to explore the effect of effective ingredients on low dose radiation protection.The mechanism of the effective components in Cibai capsule on low dose radiation protection was verified through cell cycle,apoptosis experiments,Western Blot,q-PCR,and other experiments.Result:After low dose radiation,the index of spleen and thymus in mice decreased to varying degrees.The increase of inflammatory factors(IL-1 β、IL-2、IL-6、IFN-γ)in mice serum was dose dependent,with MDA content and SOD activity in the spleen and thymus increasing or decreasing to varying degrees.White blood cells in the peripheral blood,spleen,and thymus decreased to varying degrees.B-lymphocyte and T-lymphocyte subsets in the spleen and thymus of mice were disordered,and the damage caused by radiation gradually recovered over time.Based on the above results,a low dose radiation injury model in mice was selected as the irradiation regimen of 0.2Gy for 5 consecutive days,with a cumulative dose of 1.0 Gy.Index testing was performed one day after the end of radiation.Cibai capsule can increase the total number of peripheral blood,white blood cells,and lymphocytes in irradiated mice,reduce the content of MDA in the spleen and thymus of mice,increase the activity of SOD,reduce the content of inflammatory factors in serum,regulate the subsets of B and T lymphocytes in the spleen and thymus of mice,and improve the pathological changes of the spleen of mice.The improvement effect is dose dependent.Syringa,quercetin,kaempferol,and ferulic acid can increase the total number of peripheral blood,leukocytes,and lymphocytes in highly irradiated mice to varying degrees,reduce the content of MDA and increase the activity of SOD in the spleen and thymus of mice,reduce the content of inflammatory factors in serum,and regulate the subsets of B and T lymphocytes in the spleen and thymus of mice.Syringa,quercetin,kaempferol,and ferulic acid can promote the phosphorylation of AKT in the spleen and thymus of mice,downregulate the expression of Bax and IL-6 proteins,reverse the apoptosis and inflammatory signal pathways induced by low dose radiation,and thereby promote the proliferation of spleen cells and inhibit their apoptosis.Conclusion: Cibai capsule and its effective components have a certain protective effect on spleen and thymus damage caused by low dose radiation.In addition,network pharmacology predicts that the mechanism of action of Cibai capsule is related to AKT,Bax,IL-6 and other targets.Through experimental verification,the active ingredient activated AKT can directly phosphorylate various transcription factors,which can inhibit the expression of apoptotic genes,enhance the expression of anti-apoptotic genes and promote cell survival,thereby playing a protective role against low dose radiation.