Crafts Trevor D, Hunsberger Erin Bailey, Jensen Amanda R, Rescorla Frederick J, Yoder Mervin C, Markel Troy A
Section of Pediatric Surgery, Department of Surgery, Riley Hospital for Children at Indiana University Health and, The Indiana University School of Medicine, Indianapolis, Indiana.
Section of Neonatology, Department of Pediatrics, Riley Hospital for Children at Indiana University Health and, The Indiana University School of Medicine, Indianapolis, Indiana.
J Surg Res. 2015 Dec;199(2):428-34. doi: 10.1016/j.jss.2015.06.031. Epub 2015 Jun 18.
Direct peritoneal resuscitation (DPR) has previously been shown to alter blood flow in the small bowel mesenteric vessels in models of intestinal ischemia. However, a survival advantage or its effects on local tissue inflammation have not been previously demonstrated. We hypothesized that DPR would increase survival and decrease intestinal tissue inflammation after intestinal ischemia and reperfusion (I/R) injury.
Eight-week-old male C57Bl6J mice were anesthetized and underwent midline laparotomy. I/R and DPR groups were exposed to superior mesenteric artery occlusion for 60 min with a nontraumatic clamp. Immediately after removal of the clamp, 1 mL of phosphate-buffered saline, 1 mL of minimal essential media, or 1 mL of minimal essential media supplemented with fetal bovine serum, penicillin and/or streptomycin, and glutamine were placed into the abdominal cavity of DPR groups. Animals were then closed in two layers and allowed to reperfuse for 6 h (cytokine analysis, n = 6 per group) or 7 d (survival analysis, n = 10 per group). After 6 h of reperfusion, animals were euthanized. Intestines were harvested and homogenized. Extracts were quantified for total protein content (Bradford assay), myeloperoxidase activity, tissue inflammatory cytokine, and growth factor production. P < 0.05 was significant.
I/R caused marked intestinal ischemia, significant mortality, and a significant increase in tissue cytokine and growth factor levels (P < 0.05). Seven-day survival was 30% for I/R without treatment and rose to 60% with DPR therapy using phosphate-buffered saline as the dialysate. DPR using plain MEM or MEM with supplements after ischemia increased 7-d survival to 90% (P < 0.05). DPR also significantly decreased intestinal tissue levels of myeloperoxidase, as well as intestinal tissue levels of multiple growth factors and inflammatory cytokines.
DPR increases survival and decreases intestinal inflammation after intestinal I/R injury. Translational applications are readily achievable and should be considered for patients with intestinal ischemic pathology.
先前已证明,在肠缺血模型中,直接腹膜复苏(DPR)可改变小肠肠系膜血管中的血流。然而,此前尚未证明其具有生存优势或对局部组织炎症的影响。我们推测,DPR可提高肠缺血再灌注(I/R)损伤后的生存率,并减轻肠道组织炎症。
对8周龄雄性C57Bl6J小鼠进行麻醉,并实施中线剖腹术。I/R组和DPR组使用无创夹夹闭肠系膜上动脉60分钟。移除夹子后,立即将1 mL磷酸盐缓冲盐水、1 mL基本培养基或1 mL补充有胎牛血清、青霉素和/或链霉素以及谷氨酰胺的基本培养基注入DPR组的腹腔。然后将动物分两层缝合,并使其再灌注6小时(细胞因子分析,每组n = 6)或7天(生存分析,每组n = 10)。再灌注6小时后,对动物实施安乐死。取出肠道并匀浆。对提取物进行总蛋白含量(Bradford法)、髓过氧化物酶活性、组织炎性细胞因子和生长因子产生量的定量分析。P < 0.05具有统计学意义。
I/R导致明显的肠缺血、显著的死亡率以及组织细胞因子和生长因子水平的显著升高(P < 0.05)。未经治疗的I/R组7天生存率为30%,使用磷酸盐缓冲盐水作为透析液的DPR治疗可使其升至60%。缺血后使用普通MEM或补充剂的MEM进行DPR可使7天生存率提高至90%(P < 0.05)。DPR还显著降低了肠道组织中的髓过氧化物酶水平,以及多种生长因子和炎性细胞因子的肠道组织水平。
DPR可提高肠I/R损伤后的生存率并减轻肠道炎症。转化应用很容易实现,对于患有肠道缺血性病变的患者应予以考虑。