Effect of total flavonoids from Forsythia suspensa leaf on zebrafish ulcerative colitis
This study aims to investigate the impact of total flavonoids extracted from Forsythia suspensa leaf on zebrafish ul-cerative colitis induced by 2,4,6-trinitrobenzene sulfonic acid(TNBS).The FSLF was purified using D101 macroporous res-in and its components were analyzed via UPLC-MS.Thirty-six wild zebrafish were divided into a control group,model group,low-dose FSLF group(10 μg/mL),medium-dose FSLF group(20 μg/mL),high-dose FSLF group(40 μg/mL),and dexa-methasone group(50 μg/mL).ulcerative colitis was induced in all groups except for the blank group by anal injection of 160 mmol/L TNBS.The colonic histopathology and goblet cell number of zebrafish were observed using hematoxylin-eosin and al-cian blue staining.The levels of reactive oxygen species(ROS)and superoxide dismutase(SOD)in zebrafish colon tissue were assessed using a detection kit.Fluorescence quantitative PCR detected the presence of MyD88 and TNF receptor associ-ated factor 6(TRAF6)in the colon tissue of zebrafish.Additionally,mRNA expression of nuclear factor-κB(NF-κB)p65,tumor necrosis factor-α(TNF-α),interleukin-1β(IL-1β),and interleukin-10(IL-10)was also observed.The highest con-centration of total flavonoids was found in the 30％ethanol-eluting fraction of FSLF,UPLC-MS analysis identified nine fla-vonoids,including rutin,isoquercetin,kaempferol-3-glucorhamnoside,hypericin,kaempferin-3-O-rutinoside,quercetin,Hesper-idin,kaempferol and porphyrin.Compared to the model group,FSLF demonstrates an improvement in the colonic structure of zebrafish,as evidenced by an increase in goblet cell number(P＜0.001),a decrease in ROS content(P＜0.05),downregu-lation of MyD88/TRAF6/NF-κBp65,TNF-α,IL-1β mRNA expression levels,and an increase in SOD activity(P＜0.01)and IL-10 mRNA expression levels(P＜0.05).This study indicates that FSLF exerts a significant therapeutic impact on ze-brafish ulcerative colitis,potentially through the modulation of inflammatory factors TNF-α and IL-1β,as well as the enhance-ment of IL-10 mRNA expression via regulation of the NF-κB signaling pathway.
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