大黄干预炎症小体活化调控线粒体自噬改善脓毒症全身炎症反应及保护肠道结构机制研究OA

Objective:To investigate the mechanism of rhubarb by activating the PINK1/Parkin pathway to regulate mitophagy and intervene in inflammasome activation to inhibit systemic inflammatory response and protect intestinal structure in sepsis mice.Method:A total of 40 6-8-week-old C57BL/6J male mice were randomly divided into high-dose rhubarb group,medium-dose rhubarb group,low-dose rhubarb group,sepsis model group and blank group.The high-dose rhubarb group,the medium-dose group,the low-dose group,and the sepsis model group were all modeled by cecal ligation and perforation to establish a mouse model of sepsis,After successful modeling,each dose group of rhubarb was given 0.52 mL/day of rhubarb liquid by gavage every day for 7 days,and on the 8th day,the eyeball blood of mice in each group was collected after successful modeling,serum was prepared,and intestinal tissue was retained;HE staining was used to observe the histopathological changes of intestinal tissue,and the levels of TNF-α,IL-1β and IL-6 in serum were detected by ELISA,Western blot was used to detect the expression levels of NLRP3,PINK1,Parkin,LC3I,LC3II in ileal tissues of mice in each group.Results:HE staining showed that the intestinal mucosal epithelial cell injury was the most serious in the sepsis model group,with multiple ulcers formed,obvious intestinal villi with a large amount of necrosis and large sloughing,accompanied by mucosal bleeding and edema,inflammatory cell infiltration reached the muscularis mucosa,goblet cells were significantly reduced,paneth cells were significantly increased,and inflammatory cells infiltrated into the muscularis mucosa;Compared with the model group,the intestinal mucosal epithelial cell damage,microvilli destruction and inflammatory cell infiltration in the rhubarb group were improved,and the number of goblet cells increased,while the number of paneth cells decreased,and the inflammatory cell infiltration was basically limited to the lamina propria.The serum TNF-α,IL-1β,IL-6 level in the sepsis model group was significantly increased(P<0.01),and the serumTNF-α,IL-1β,IL-6 level in the rhubarb group decreased after treatment(P<0.05,P<0.01).The expression levels of NLRP3,PINK1,Parkin,LC3I,LC3II were significantly increased in the intestinal tissues of mice in the sepsis model group(P<0.01).After treatment,the level of NLRP3 in each dose group of rhubarb decreased(P<0.01),which was negatively correlated with the dose,and the expression level of PINK1,Parkin,LC3I,LC3II increased to varying degrees(P<0.05,P<0.01),which was positively correlated with the dose.Conclusion:By inhibiting the activation of intestinal NLRP3 inflammasomes,rhubarb activates the PINK1/Parkin pathway and increases the degree of mitophagy,thereby reducing the production of systemic inflammatory factors and protecting the intestinal structure.