Experimental Study On Protective Effect Of Ydrogen-rich Saline On Intestine Damage In Scalded Ats Following Delayed Resuscitation

Background and objectiveLarge number of body fluids oozing or amass in the tissue space after severe burn resulting inhypovolemia and even shock. The intestine is the most earlist and vulnerable organ to ischemiadue to blood flow redistribution which is regulated by the nervous and endocrine systems.Sometime after ischemia,xanthine dehydrogenase is converted to xanthine oxidase due to the role ofcalcium in the epithelial cells.During reperfusion, molecular oxygen is reintroduced into thetissue, where it reacts with hypoxanthine or XO to produce a burst of reactive oxygenspecies.Excessive ROS damage the intestinal mucosa and the barrier, thus bacteria and toxins iseasy to shift.This result systemic infection,systemic inflammatory response syndrom(SIRS) andeven multiple organ dysfunction syndrome (MODS).Infection and MODS currently is themost important cause of death in burn patients.The role of hydrogen saline in scavengingreactive oxygen species have a potential application, and may reduce oxidative stress and therelease of inflammatory mediators, and to protect intestinal and distant organ. The aim ofthis study is to investigate whether hydrogen-rich saline exerts intestine oxidative damageprotection against severe burn.MethodsForty-eight rats were divided into four groups: sham plus normal saline(S,n=12),burn injuryplus normal saline(B+N,n=12), burn injury plus hydrogen-rich saline(B+H,n=12), and burninjury plus vitami C(B+C,n=12). Animals were subjected to full-thickness burn wound (30%total body surface area) using boiling water(98℃,12S).Lactated Ringer’s solution was given at6h post-burn. The rats in hydrogen group received5ml/kg of hydrogen-rich saline, sham and burncontrols obtained the same amount of saline, and the VitC group was treated with 9ml/kg(250mg/kg) of VitC in saline at6,18,30,42hour post scalding by intraperitoneal injectionrespectively.6rats of each group were put to death and then we collected intestine tissue todetect related indicators at24,48hour post scalding respectively.1.Observe the changes ofintestinal pathological （H&E staining）,and then evaluate the intestine tissue injury andintestinal mucosal damage;2. intestinal tissue malondialdehyde (Malondialdehyde, MDA) andIL-1beta content, comparing the groups of degree of oxidative damage and inflammatorychanges;3. intestinal tissue DAO (diamine oxidase, DAO) content, evaluate rat intestinalmucosal integrity of each group;4. intestinal villous epithelial cell apoptosis rate, evaluateintestinal epithelial cell apoptosis of each group.Results1.Delayed resuscitation after severe burns can cause the intestinal tissue injury in rats. Changesof intestinal mucosa pathological can be found such as villi shorter,the intestine epithelial celldegeneration, necrosis, shedding,vills gap widened. The application of hydrogen-rich saline andvitamin C can improve the degree of damage of the intestinal mucosa.Intestinal pathologicalwere damaged significantly heavier at48hour post scalding than the24h (P <0.01).Hydrogen-rich saline and vitamin C treatment groups were no significant difference for theimprovement of the intestinal pathological at24,48hour post scalding.2. Compared with S group, the intestinal tissue malonylaldehyde(MDA) of B+N group wassignificantly increased (P<0.01), while both B+H group and B+C group is significantly lessthan B+N group (p <0.01).MDA content peaked at24hour post scalding.And effect of vitaminC treatment group is more protective than hydrogen-rich saline at48hour post scalding(P<0.05).3. Compared with S group,intestinal tissue IL-1β expression of B+N group was significantlyincreased (P<0.01).Expression of intestinal tissue IL-1β peaked at48hour post scalding.Both hydrogen-rich saline and vitamin C treatment group significantly reduce expression ofinflammatory cytokines (B+H vs. B+N, p <0.01; B+C vs. B+N, P<0.01).Effect ofhydrogen-rich saline treatment group is more protective than hydrogen-rich saline at24hourpost scalding(P <0.05).4.From the result of DAO content test, intestinal tissue DAO content of B+N group was significantly reduced compared with S group (P <0.01). DAO content peaked at48hour postscalding. Both hydrogen-rich saline and vitamin C treatment group significantly increased DAOcontent (B+H vs. B+N,P <0.01; B+C vs. B+N, P<0.01).And DAO content of hydrogen-richsaline is increased than vitamin C at48hour post scalding(P <0.05).5. Compared with S group, the rate of apoptosis of intestinal villus epithelial cells of B+N groupis significantly higher (P <0.01), hydrogen-rich saline and vitamin C can significantly reducethe rate of apoptosis of intestinal villus epithelial cells.(B+H vs. B+N, P <0.01; B+C vs. B+N, P<0.01). The rate of apoptosis of intestinal villus epithelial cells peaked at24hour postscalding. Hydrogen-rich saline and vitamin C treatment groups were no significant differencefor the improvement of the rate of apoptosis of intestinal villus epithelial cells at24,48hourpost scalding.ConclusionsHydrogen-rich saline can reduce oxidative stress to protect intestine damage which is caused bydelayed resuscitation after severe burns.