The role regenerating islet-derived protein 3βin intestinal repair and lactylation mechanisms in septic mice
10.3969/j.issn.1008-9691.2025.01.007
- VernacularTitle:胰岛再生蛋白3β对脓毒症小鼠肠道的修复作用及乳酸化机制研究
- Author:
Xihua ZHEN
1
;
Zexian SUN
;
Yonghao YU
;
Xin WANG
Author Information
1. 天津医科大学总医院麻醉科,天津 300052
- Publication Type:Journal Article
- Keywords:
Sepsis;
Regenerating islet-derived protein 3β;
Intestinal barrier;
Lactylation;
Macrophage
- From:
Chinese Journal of Integrated Traditional and Western Medicine in Intensive and Critical Care
2025;32(1):37-42
- CountryChina
- Language:Chinese
-
Abstract:
Objective To investigate the effects of regenerating islet-derived protein 3β(Reg3β)on intestinal function and glycolysis in septic mice,as well as its role in promoting lactylation.Methods ① In vivo experiments:a total of 36 adult male C57BL/6 mice,including wild-type(WT)and Reg3β knockout(KO)mice,were randomly divided into six groups using a random number table:WT sham group,WT cecal ligation and puncture(CLP)-induced sepsis group(WT CLP group),WT sham+Reg3β intervention group(WT sham group),WT CLP+Reg3β intervention group(WT CLP+Reg3β group),KO sham group,and KO CLP group(n=6 per group).Blood glucose levels were measured at 24 hours and 48 hours after modeling;At 48 hours after modeling,ileum tissues were collected for hematoxylin-eosin(HE)staining to observe histopathological changes,immunofluorescence staining was performed to assess the positive expression levels of lactylated proteins,Western blotting was used to detect the expression levels of lactylated proteins in ileum tissues.② In vitro experiments:Cultured RAW264.7 cells were randomly divided into four groups using a random number table:blank control group,lipopolysaccharide(LPS)-induced sepsis model group(LPS group),Reg3β group,and LPS+Reg3β group.After 24 hours of drug induction,cells were collected,and Western blotting was performed to measure the levels of lactylated proteins,the culture medium was collected to determine lactylation levels.Results ① Histopathological observations showed that compared with the WT CLP group,the WT CLP+Reg3β group exhibited milder villus breakage and inflammatory cell infiltration.The KO CLP group showed more severe damage,with significantly shortened intestinal villi and separation of the epithelial layer from the lamina propria.Compared with the WT CLP group,blood glucose levels were significantly higher in the KO CLP group(mmol/L:6.83±1.15 vs.4.78±1.37,P<0.05).Both Western blotting and immunofluorescence staining results indicated that,compared with the WT CLP group,lactylation levels were significantly decreased in the KO CLP group[lactylated protein expression(lactylated protein/β-actin):0.48±0.20 vs.0.78±0.09;positive lactylated protein expression(mean fluorescence intensity):59.84±6.02 vs.100.00±5.26,both P<0.01].② Western blotting results of RAW264.7 cells cultured for 24 hours showed that compared with the LPS group,the LPS+Reg3β group exhibited significantly increased lactylated protein expression levels(lactylated protein expression/β-actin:3.67±0.48 vs.1.64±0.49,P<0.01).Compared with the blank control group,the lactate levels in the culture medium of the LPS group were significantly increased(mmol/L:4.95±0.20 vs.3.82±0.09,P<0.01).Compared with the LPS group,the lactate levels in the culture medium of the LPS+Reg3β group were also significantly increased(mmol/L:6.03±0.32 vs.4.95±0.20,P<0.01).Conclusion Reg3β promotes intestinal protein lactylation and exerts a protective effect on the intestine in sepsis,suggesting that Reg3β may serve as a novel therapeutic target for sepsis.
