| Chicken proventriculitis is a prevalent infectious disease,resulting in huge economic losses to the global poultry industry.The etiology of proventriculitis is quite complex,bringing trouble to the diagnosis and treatment of proventriculitis itself.Characterized by glandular stomach enlargement,chicken proventriculitis causes serious damage to the gastric mucosa.Oxidative stress is a significant driver of gastrointestinal diseases including gastric mucosal injury.Pomegranate peel contains numerous phenolic compounds with various biological functions,especially the antioxidant effect,thereby having high nutritive and medicinal values.Dahuang Sanxian Powder is composed of pomegranate peel,rhubarb charcoal,fried Divine Comedy,fried malt and hawthorn charcoal,in which pomegranate peel is the monarch drug.Dahuang Sanxian Powder has the functions of improving digestion,invigorating stomach,activating blood and resolving stasis,and stopping diarrhea,which may have a therapeutic effect on chicken proventriculitis.Punicalagin is the most abundant and active polyphenolic compound in pomegranate peel with strong antioxidant activity,and has been shown to have protective effects against gastric injury.Therefore,this study aimed to explore the antioxidant effect and mechanism of Dahuang Sanxian Powder and punicalagin on the gastric mucosal injury in chicken proventriculitis,and the detailed research contents and results are as follows:First,we explored the therapeutic effect of Dahuang Sanxian Powder on lipopolysaccharide(LPS)-induced chicken proventriculitis.We established a chicken proventriculitis model induced by 5 μg/bird LPS via intragastric administration.The chicks in the model group brought about diarrhoea,decreased body weight and increased glandular stomach index.Macroscopic lesions included intestinal gas and bleeding,mucosal erosion,swollen and flat glandular papilla and gizzard cuticle necrosis.Histopathological observation included the local necrosis and shedding of the glandular stomach mucosa epithelium and glandular epithelium.These characteristics in the model group were consistent with the clinical symptoms and pathological characteristics of chicken proventriculitis,indicating that the LPS-induced chicken proventriculitis model was successfully established.1 g/kg,2g/kg or 4 g/kg Dahuang Sanxian Powder were taken by intragastric administration for five days,clinical symptoms were observed after withdrawal for five days,and after the chicks were euthanized,the glandular stomach and other tissues were collected for pathological observation,and the secretion and activity of prostaglandin E2(PGE2)and superoxide dismutase(SOD)in the glandular stomach tissue were detected.The results showed that compared with the model group,in the treatment groups the body weight was increased,the glandular stomach index was decreased,duodenal bleeding,glandular papilla swelling and mucosal erosion was alleviated,the local necrosis and inflammation infiltration of glandular epithelium was mitigated,and the secretion and activity of PGE2 and SOD in the glandular stomach tissue were improved.These results suggested that Dahuang Sanxian Powder had a significant therapeutic effect on LPS-induced chicken proventriculitis,which may be related to its antioxidant activity.In order to further explore the mechanism of Dahuang Sanxian Powder on chicken proventriculitis,we explored the antioxidant effect and mechanism of punicalagin,the main component of pomegranate peel which is the monarch drug in Dahuang Sanxian Powder,on LPS-induced chicken glandular gastric epithelial cells.The chicken glandular stomach epithelial cells were stimulated by 4 μg/ml LPS for24 h,and treated with 4 μM,8 μM or 16 μM punicalagin for 24 h,and then the cell damage,oxidative stress,gastric mucosal protective factor expression,tight junction protein expression and Nrf2 signaling pathway activation were detected.The results showed that after LPS treatment,the cell viability of glandular gastric epithelial cells was significantly decreased and the LDH release was significantly increased;the ROS level was significantly increased,and total SOD and total GSH were significantly decreased;gastric mucosal protective factors PGE2 and NO levels and TFF2 m RNA expression were significantly decreased;tight junction proteins ZO-1,Occludin and Claudin-3 m RNA expression were significantly decreased;the expression of Nrf2 signaling pathway-related proteins,including Nrf2,NQO1 and Keap1,were significantly decreased.Punicalagin could significantly reverse these changes induced by LPS in chicken glandular gastric epithelial cells.After punicalagin treatment,the cell viability of chicken glandular gastric epithelial cells was significantly increased and the LDH release was significantly decreased;the ROS level was significantly decreased,and total SOD and total GSH were significantly increased;the secretion of PGE2 and NO and the m RNA expression of TFF2 were significantly improved;the m RNA expression of ZO-1,Occludin and Claudin-3 were significantly increased;the expression of Nrf2,NQO1 and Keap1 were significantly increased.The Nrf2 inhibitor,ML385,significantly inhibited the antioxidant effect of punicalagin on LPS-induced chicken glandular gastric epithelial cells.After ML385 treatment,the ROS level was significantly increased,and the m RNA expression of ZO-1,Occludin and Claudin-3were significantly decreased.In conclusion,Dahuang Sanxian Powder had a therapeutic effect on LPS-induced chicken proventriculitis,significantly increased body weight and reduced glandular stomach index,and alleviated the clinical symptoms and pathological lesions of LPS-induced chicken proventriculitis,and the therapeutic effect may be related to punicalagin,the main component of pomegranate peel which is the monarch drug in Dahuang Sanxian Powder.Punicalagin had an antioxidant protective effect on LPS-induced chicken glandular gastric epithelial cells,significantly reduced oxidative stress,and promoted the expression of gastric mucosal protective factors and tight junction proteins,and the antioxidant effect of punicalagin was achieved by regulating the activity of Nrf2 signaling pathway.Our research would provide new ideas and theoretical basis for the prevention and treatment of chicken proventriculitis,and offer positive promotion of significance to the healthy development of the poultry industry. |
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