Alleviative Effects Of Interference With SLC6A14 Against Ulcerative Colitis Via Inhibiting NLRP3 Inflammasome

Objective: Ulcerative colitis(UC)is a kind of colorectal inflammation with unknown etiology.The incidence of UC is increasing in China,and its treatment is facing challenges.Studies have confirmed that the expression of Solute carrier family 6 member 14(SLC6A14)is up-regulated in UC,but its role in the pathogenesis of UC has not been elucidated.Nucleotide binding oligomerization domain-like receptor protein 3(NLRP3)inflammasome is activated in UC and is closely related to UC.It is unknown whether SLC6A14 can affect the progression of UC by regulating NLRP3 inflammasome.This study explored the mechanism of SLC6A14 on the occurrence and development of UC based on NLRP3 inflammasome.Methods: In this study,the mechanism of knockdown of SLC6A14 inhibiting the activation of NLRP3 inflammasome and antagonizing UC was investigated at the cellular and animal levels.The first step was to culture human normal intestinal epithelial cells,construct SLC6A14 interference plasmid,transfect cells,and use LPS to construct an inflammatory cell model.CCK-8 and Ed U staining were used to detect cell proliferation;Apoptosis was detected by flow cytometry;the expression levels of ASC,Pro-IL-1β,IL-1β,Pro-IL-18,IL-18,NLRP3 and SLC6A14 were detected by Western blot;The levels of IL-1β and IL-18 in the supernatant were detected by ELISA;The expression level of SLC6A14 was detected by q PCR.In the second step,on the basis of the first step,SLC6A14 interference plasmid and NLRP3 overexpression plasmid were constructed and transfected into cells.The same method was used to detect the changes of cell proliferation and apoptosis,and the same indicators as the first step.Finally,an animal model of colitis was established,and SLC6A14 interference lentivirus was used for intervention.Observe and record the physiological state of mice;HE staining was used to observe the structure of colon tissue and the degree of inflammatory cell infiltration.The same method was used to detect changes in the same indicators as the first step.Results: 1.Interference with SLC6A14 inhibits NLRP3 inflammasome activation to antagonize cell inflammation.Compared with the control group,LPS group had less cell proliferation,more apoptosis,and up-regulated SLC6A14 and NLRP3 inflammasome related indicators;compared with the LPS group,after interfering with SLC6A14,SLC6A14 expression was down-regulated,cell proliferation was more,apoptosis was less,and NLRP3 inflammasome-related indicators were down-regulated.2.The activation of NLRP3 inflammasome can reduce the protection of SLC6A14 on inflammation and promote cell inflammation.After overexpression of NLRP3 in SLC6A14-interfering cells,compared with the SLC6A14-interfering group,cell proliferation was less,apoptosis was more,and NLRP3 inflammasome-related indicators were up-regulated.3.Interference with SLC6A14 affects the activation of NLRP3 inflammasome and antagonizes colitis in mice.Compared with the control group,the body weight of mice in the DSS group was significantly decreased,the Disease activity index(DAI)score was significantly increasedthe pathological damage of the colon was aggravatedand SLC6A14 and NLRP3 inflammasome-related in dicators were up-regulated.Compared with DSS group,after SLC6A14 intervention,th e weight of mice increased,the survival rate increased,the DAI score decreased,the p athological damage of colon was alleviated,and SLC6A14 and NLRP3 i nflammasome-related indicators were down-regulated.Conclusion: The increased expression of SLC6A14 is closely related to the occurrence and development of UC.SLC6A14 participates in UC by activating NLRP3 inflammasome.