A Study On The Intervention Effect Of Total Saponin From Panax Japonicus C.A.Mey On Kawasaki Disease Based On The TLR4/NF-ΚB Signaling Pathway

Background: Kawasaki disease occurs in infants under 5 years of age,and its incidence is increasing year by year.It has become the primary cause of acquired heart disease in children.It is an urgent problem to find specific experimental indicators for early diagnosis of KD to improve the timeliness and accuracy of disease diagnosis and to give effective treatment.Objective This study was based on TLR4/NF-κB signaling pathway to investigate the intervention effect of total saponins from Panax japonicus C.A.Mey on Kawasaki disease.Methods The BALB/c male mice aged 4-6 weeks were randomly divided into 4 groups,namely normal control group,KD model group,low-dose group and high-dose group of total saponins from Panax japonicus C.A.Mey,with 10 mice in each group and the body weight of each mouse was about 15-20 g.The mice in normal control group were intraperitoneally injected 0.5 m L of PBS solution,and mice in KD model group,low-dose group and high-dose group of total saponins from Panax japonicus C.A.Mey were intraperitoneally injected 0.5 m L of LCWE solution,respectively.From the 4th day after modeling,different concentrations of total saponins from Panax japonicus C.A.Mey in the low-dose and high-dose groups of total saponins of Panax japonicus C.A.Mey were given intragastric administration.Mice in normal control group and model group were given the same amount of PBS intragastric administration for 10 days.(1)During feeding period,the general condition of mice was observed and the changes in body weight of mice were recorded.(2)The inflammation of the coronary arteries in the hearts of mice were stained with hematoxylin and eosin(HE).(3)The relative m RNA levels of TLR4,My D88,NF-κB p65 and TNF-α in heart tissues of mice in each group were determined by Real-time PCR.(4)The relative protein levels of TLR4,My D88,p-NF-κB p65 and TNF-α in heart tissues of mice in each group were determined by Western blot.Results(1)The mouse model of Kawasaki disease was successfully established.(2)The body weight of mice in the model group increased more slowly than that in the control group.After intervention treatment of total saponins from Panax japonicus C.A.Mey,the body weight of mice in the low-dose group of total saponins of Panax japonicus C.A.Mey increased compared with that in the model group,while the body weight of mice in the high-dose group of total saponins of Panax japonicus C.A.Mey increased more obviously than that in the model group.(3)The results of H&E staining showed that the infiltration of inflammatory cells around the coronary arteries of mice in the KD model group was more than that in the control group,and the coronary arteries were thickened and the inner wall of the vessels was less smooth.After the intervention of low dose of total saponins from Panax japonicus C.A.Mey,coronary artery inflammation was reduced,while after the intervention of high dose of total saponins from Panax japonicus C.A.Mey,the improvement of coronary artery inflammation was more obvious.(4)The relative m RNA levels of TLR4,My D88,NF-κB p65 and TNF-αin heart tissue of mice in KD model group were significantly higher than those in normal control group.Compared with KD model group,The relative m RNA levels of TLR4,My D88,NF-κB p65 and TNF-αin heart tissues of mice in low and high dose groups of total saponins from Panax japonicus C.A.Mey had different degrees of reduction.(5)The relative protein levels of TLR4,My D88,p-NF-κB p65 and TNF-α in heart tissue of mice in KD model group were significantly higher than those in normal control group.Compared with KD model group,The relative protein levels of TLR4,My D88,p-NF-κB p65 and TNF-α in heart tissues of mice in low and high dose groups of total saponins from Panax japonicus C.A.Mey had different degrees of reduction.Conclusion(1)The TLR4/NF-κB signaling pathway was activated and the expression of related factors was increased in Kawasaki disease model mice induced by intraperitoneal injection of LCWE.(2)The total saponins of Panax japonicus C.A.Mey have therapeutic effect on Kawasaki disease,possibly through inhibiting the activation of TLR4/NF-κB signaling pathway.