Bacterial Metabolites In Hyperuricemia Regulate The Intestinal Barrier Through The PKC Pathway

Research background Gout is a metabolic disease related to hyperuricemia caused by disorder of purine metabolism or decreased excretion of uric acid.Due to the increased production of uric acid and decreased excretion,the typical clinical symptom of gout is hyperuricemia.Hyperuricemia is closely related to hypertension,hyperlipidemia,cardiovascular and cerebrovascular diseases and other related metabolic diseases.One-third of uric acid in healthy people is excreted through the intestine.Intestinal microbial metabolism disorder and intestinal barrier function are influencing factors that cause various metabolic diseases,and microbial metabolism affects the intestinal barrier function to a certain extent.Research purposes Hyperuricemia can lead to increased intestinal permeability.It has been confirmed in previous studies that increased intestinal permeability is closely related to the occurrence of various metabolic diseases.Increased intestinal permeability has been found in diabetes,fatty liver,obesity and other diseases.A large number of related mechanisms have been studied,but the mechanism of increased intestinal permeability in hyperuricemia is currently unclear.Early sequencing results found that PKC changes significantly.By exploring the influence of PKC signaling pathway on the intestinal barrier,it can provide new possibilities for the mechanism of increased intestinal permeability of hyperuricemia.Research methods 1.The colon tissues of C57BL/6 wild-type mice and Uox gene knockout hyperuricemia model mice(Uox-/-)were collected,and Western Blot was used to detect PKC and MLC in the colon tissues of the two groups of mice.,MLCK,the expression changes of PPKC,PMLC,PMLCK and the tight junction protein complex Occludin and Claudin-1 at the phosphorylation level.2.The colon cancer cell Caco-2 was used as a model,and no treatment was used as a blank control group.Normal mouse feces and gout model mouse fecal supernatants were used to stimulate cells to simulate in vivo experiments,and the tight junction protein complex and PKC signaling pathway were measured respectively.Variety.In order to further verify whether PKC plays a key role in regulating intestinal permeability,PKC inhibitors were further added to measure the changes of tight junction protein complex and PKC signaling pathway.Results The protein levels of tight junction protein macromolecules Occludin and Claudin-1in the colon tissue of hyperuric acid mice were lower than those of wild-type mice,while the protein levels of tight junction protein small molecules Occludin and Claudin-1 increased(P<0.05).The protein levels of phosphorylated PMLCK,PMLC,and PPKC in hyperuricemia mice were significantly higher than those of wild-type mice,and their phosphorylation levels were significantly higher than those of wild-type mice(P<0.05).In the simulated in vitro experiment,Caco-2 cells were stimulated with high uric acid mouse fecal supernatant for 24 hours,the phosphorylated PPKC,PMLC,PMLCK protein expression levels were significantly higher than the blank control group and wild-type mouse fecal supernatant stimulation group,The phosphorylation level was significantly increased(P<0.05).After adding PKC inhibitor,the expression of PKC was suppressed,and the depolymerization of tight junction protein macromolecular complex was inhibited to a certain extent(P<0.05).Conclusion and significance The metabolites of the high uric acid mouse flora regulate the intestinal barrier through the PKC signaling pathway,which depolymerizes tight junction protein macromolecules,resulting in increased expression of tight junction protein small molecule protein levels and increased intestinal permeability.By inhibiting the expression of the PKC gene,it is concluded that the expression of tight junction protein is contrary to the previous results.It can be seen that PKC is involved in the regulation of the intestinal barrier to a certain extent,and the inhibition of PKC can inhibit the increase in intestinal permeability to a certain extent.The mechanism of increased intestinal permeability caused by hyperemia has certain clinical significance.Research on the mechanism of intestinal permeability of hyperuricemia can provide a new perspective for the future treatment of hyperuricemia and its complications.