| Selenomethionine(SeMet)is one of the main selenium sources for cultured animals,which has many protective effects such as anti-inflammatory and anti-oxidation.However,the preventive and therapeutic effect of SeMet on liver injury in laying hens is still unclear.It is known that lipopolysaccharide(LPS)can cause inflammation.In order to explore the possible protective effect of SeMet on chicken liver tissue,on the basis of replicating the model of chicken liver injury induced by LPS,we constructed the interaction model between SeMet and LPS by adding SeMet to the diet.At the same time,chicken hepatocyte line(LMH)was used to construct the in vitro validation model.The histopathological damage was observed by transmission electron microscopy,H&E,Sirius red and Masson staining.Oxidative stress level was detected by oxidative stress kit and ROS fluorescence labeling,and immunofluorescence was used to detected hepatocyte pyroptosis.Finally,using q RT-PCR and Western-Blot to detect the expressions of inflammation,pyroptosis,fibrosis and heat shock protein related genes.The results were as follows(1)The liver appearance and H&E staining in NC group and SeMet group were normal,while in LPS group,there were more punctate bleeding and necrotic lesions,degeneration and necrosis of hepatocytes,inflammatory cell infiltration,collagen fiber deposition,liver function related indexes T-BIL,ALP,CHOL,AST and ALT increased significantly,while the changes and injuries were noteworthy improved in LPS+SeMet group(P<0.05).The mRNA expressions of inflammation-related factors TNF-α,IL-6 and IL-1β were markedly up-regulated in LPS group,while IL-2 was observable down-regulated compared with NC group(P<0.05),fibrosis-related genes(α-SMA,TGF-β and Vimentin)were signally enhanced and E-cadherin was signally descended in mRNA and protein expressions,while SeMet coprocessing could observable relieved(P<0.05).These results confirmed that SeMet could alleviate LPS-induced liver inflammation and fibrosis.(2)The results of CAT,SOD,GSH,MDA and ROS oxidative stress-related indicators showed that compared with the NC group,SeMet group antioxidant capacity was signally enhanced,while the LPS group oxidative stress level was significantly advanced accompanied by ROS accumulation(P < 0.05).The above changes in the LPS+SeMet group were memorable alleviated(P < 0.05).The results of HSP60,HSP70 and HSP90 detection in mRNA and protein expression showed that LPS expose could induce these genes expression enhancement,and SeMet could significantly inhibit them after interacting with them(P<0.05).Cell tests also confirmed the above trends.The above results confirmed that SeMet could alleviate LPS-induced oxidative stress and over-expression of heat shock protein in liver tissue.(3)The results of NLRP3,GSDMD-N’,GSDMA,GSDME,ASC,Caspase1,IL-1β and pro-IL-1βpyroptosis-related genes showed that,there was no remarkable difference in mRNA and protein level expression between the NC group and SeMet group(P > 0.05),but it was significantly increased in LPS group.Immunofluorescence detection showed that the expression levels of NLRP3 and GSDMD in the LPS group were increased,while the above expression changes were significantly relieved after interaction between SeMet and LPS(P < 0.05).The above results confirmed that SeMet could alleviate LPS-induced pyroptosis of liver tissue cells.To sum up,dietary SeMet can effectively alleviate the increase of heat shock protein expression and oxidative stress in the liver of laying hens induced by LPS,and can improve the liver injury induced by LPS by inhibiting the inflammation,fibrosis and pyroptosis of liver tissue.The results of this study can provide nutritional reference for poultry breeding and feed development,and provide new ideas for poultry disease treatment. |
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