Potential influences of expression levels of MFGE8 and HMGB1 on the intestinal mucosal barrier function and inflammatory response after blunt abdominal injury in ratsPurpose: To explore the influence of milk fat globule-EGF factor 8 protein (MFGE8) on blunt abdominal injury in Sprague Dawley (SD) rats through the RhoA/ROCK signaling pathway.
Methods: The blunt abdominal injury model was generated in SD rats. A total of 44 rats was randomly assigned into three groups. Rat blunt abdominal injury was assessed by the abbreviated injury scale (AIS). The rats were sacrificed for observing the morphology of the abdominal cavity and intestines. Hematoxylin and eosin staining was performed to visualize the pathological changes of rat intestines. Positive expressions of MFGE8 and high mobility group box 1 (HMGB1) in rat intestines were examined by immunohistochemical staining. Protein levels were determined by Western blot. Serum levels of tumor necrosis factor α (TNF-α), IL-1β, IL-6 and malondialdehyde (MDA) were measured by enzyme linked immunosorbent assay (ELISA).
Results: Blunt abdominal injury resulted in inflammatory response of intestinal tissues, increased serum levels of TNF-α, IL-1β, IL-6 and MDA, upregulation of HMGB1, RhoA and ROCK2, and downregulation of MFGE8 in rats, which were significantly alleviated by intervention of rhMFGE8.
Conclusions: MFGE8 protects the intestinal mucosal barrier function after blunt abdominal injury in rats by downregulating HMGB1. Moreover, it alleviates inflammatory response and oxidative stress caused by blunt abdominal injury in rats through downregulating RhoA and ROCK.
Key words
Abdominal Injuries; rho-Associated Kinases; HMG-Box Domains; Rats
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MDA: malondialdehyde; TNF-α: tumor necrosis factor α; ELISA: enzyme linked immunosorbent assay; *p < 0.05, control group vs. the remaining groups;#p < 0.05, model group at 4 h vs. model + rhMFGE8 group; &p < 0.05, model group at 24 h vs. model + rhMFGE8 group.
HMGB1: high mobility group box 1; GAPDH: Glyceraldehyde-3-Phosphate Dehydrogenase;; MFGE8: milk fat globule-EGF factor 8 protein; *p < 0.05, control group vs. the remaining groups; #p < 0.05, model group at 4 h vs. model + rhMFGE8 group; &p < 0.05, model group at 24 h vs. model + rhMFGE8 group.
HMGB1: high mobility group box 1; MFGE8: milk fat globule-EGF factor 8 protein; *p < 0.05, control group vs. the remaining groups;#p < 0.05, model group at 4 h vs. model + rhMFGE8 group; &p < 0.05, model group at 24 h vs. model + rhMFGE8 group.