Therapeutic Effect Of Artemisia Argyi On Enteritis In Zebrafish

Objective: Animal enteritis is a inflammatory disease regulated by a variety of mechanisms,which is characterized by mucosal and deep tissue necrosis,local tissue congestion,bleeding,and a large number of inflammatory cells infiltration.In recent years,the incidence rate of animal enteritis has increased year by year.Therefore,the etiology,pathogenesis and development and utilization of animal enteritis have become a hot topic of many scholars at home and abroad.Artemisia argyi is a common Chinese herbal medicine and has many pharmacological effects including anti-inflammatory,antioxidant,antibacterial.However,there are few reports on the treatment of animal enteritis with Artemisia argyi.Methods: In order to explore the therapeutic effect of Artemisia argyi on zebrafish enteritis,zebrafish were randomly divided into control group(CK),ethanol control group(EtOH),model group(Oxa),treatment group(LA,MA,HA)and positive group(Van).Zebrafish in CK group were injected physiological saline solution.Oxazolone(dissoloved in 50% ethanol solution)was injected into the rectum of zebrafish in Oxa,treatment groups,and Van group.Zebrafish in EtOH group were injected with the same amount of 50% ethanol.After 5 h of oxazolone injection,the zebrafish in treatment groups and Van group were treated with different concentrations of Artemisia argyi decoction(0.03 g/L,0.06 g/L,0.12 g/L)and vancomycin(10 mg/L),respectively.After 5 h,3 d and 6 d,the intestinal tract was randomly taken for histomorphological observation by HE staining and AB-PAS staining,for detection of the immune related enzymes and oxidative stress related indicators by reagent test kit,for the relative levels of inflammatory factors and genes in MyD88/TRAF6/NF-KB pathway by real-time fluorescent quantitative PCR to explore whether the inflammatory model was successfully established and to clarify the therapeutic effect of Artemisia argyi on zebrafish enteritis.Results:(1)After the injection of oxazolone in rectum for 5 h,compared with CK group,the intestinal mucosal structure of zebrafish in EtOH group and Oxa group was seriously damaged,including seriously exfoliated intestinal epithelial cells,infiltrated inflammatory cells,exhausted goblet cells,thickened intestinal submucosa and lamina propria,and increased acidic mucin(P< 0.05).The activities of CAT,SOD,ACP and AKP were significantly decreased(P< 0.05),while the contents of MDA,ROS and MPO activity were significantly increased(P< 0.05).The relative expressions of IL-10 mRNA was decreased,while the relative expressions of IL-1β,MyD88,TRAF6 and P65 mRNA were increased significantly(P< 0.05).Compared with female zebrafish,the change in male zebrafish was more obvious than that in female zebrafish.(2)After 3 days of treatment with Artemisia argyi decoction and vancomycin,compared with CK group,the intestinal mucosa of male and female zebrafish in Oxa group was damaged,intestinal epithelial cells fell off seriously,inflammatory cells infiltrated,and acidic mucin increased significantly;the activities of CAT,SOD,ACP and AKP decreased,and the activities of MDA,ROS and MPO decreased(P < 0.05);the relative expression of IL-10 mRNA decreased,and the levels of IL-1β,MyD88,TRAF6 and P65increased(P< 0.05).In female zebrafish,compared with Oxa group,the activities of CAT,SOD,ACP and AKP in Van group were significantly increased,while the activities of MDA and ROS were significantly decreased(P< 0.05);the relative expression of IL-10 mRNA was increased,while the relative expression of IL-1β,MyD88,TRAF6 and P65 mRNA was significantly decreased(P < 0.05).In male zebrafish,compared with Oxa group,the activities of CAT,SOD,ACP and AKP in HA group and Van group were significantly increased,while the activities of MDA and ROS were decreased(P < 0.05);the relative expression of IL-10 mRNA was increased,while the relative expression of IL-1β,MyD88,TRAF6 and P65 mRNA was significantly decreased(P < 0.05).(3)After 6 days of treatment with Artemisia argyi decoction and vancomycin,compared with CK group,the intestinal mucosa of male and female zebrafish in Oxa group was damaged,intestinal epithelial cells fell off,inflammatory cells infiltrated,and the number of acidic mucin increased significantly;the activities of CAT,SOD,ACP and AKP decreased,and the activities of MDA,ROS and MPO increased(P <0.05);the relative expression of IL-10 mRNA decreased,and the levels of IL-1β,MyD88,TRAF6 and P65 relative expression of mRNA was increased(P < 0.05).In female zebrafish,compared with Oxa group,the activities of CAT,SOD,ACP and AKP in HA group and Van group were significantly increased,while the activities of MDA and ROS were significantly decreased(P < 0.05);the relative expression of IL-10 mRNA was increased,while the relative expression of IL-1β,MyD88,TRAF6 and P65 mRNA was significantly decreased(P < 0.05).In male zebrafish,compared with oxa group,the activities of CAT,SOD,ACP and AKP in Ma group,HA group and Van group were significantly increased,while the activities of MDA and ROS were decreased(P < 0.05);the relative expression of IL-10 mRNA was increased,while the relative expression of IL-1β,MyD88,TRAF6 and P65 mRNA was significantly decreased(P < 0.05).Conclusion: oxazolone(50% ethanol solution)was injected into the rectum of male and female zebrafish to establish enteritis model.For female zebrafish,adding high-dose Artemisia argyi decoction and vancomycin can effectively improve the intestinal pathological damage,slow down the intestinal oxidative stress,repair the intestinal immune ability,inhibit the expression of IL-1β and MyD88/TRAF6/NF-KB pathway related genes and promote the expression of IL-10,thus alleviating zebrafish enteritis;for male zebrafish,adding medium dose and high-dose Artemisia argyi decoction can effectively alleviate zebrafish enteritis Conclusion Artemisia argyi decoction and vancomycin can effectively relieve zebrafish enteritis.This study can provide a theoretical basis for further study on the rationality of Artemisia argyi replacing feed antibiotics.