Khaneki Sina, Jensen Amanda R, Drucker Natalie A, Markel Troy A
Section of Pediatric Surgery, Department of Surgery, Indiana University School of Medicine, Riley Hospital for Children and Indiana University Health, Indianapolis, Indiana.
Section of Pediatric Surgery, Department of Surgery, Indiana University School of Medicine, Riley Hospital for Children and Indiana University Health, Indianapolis, Indiana.
J Surg Res. 2017 Jun 1;213:274-280. doi: 10.1016/j.jss.2017.02.060. Epub 2017 Mar 4.
Direct peritoneal resuscitation (DPR) has been shown to increase survival after intestinal ischemia and reperfusion injury (I/R). We have previously appreciated that minimum essential medium (MEM), a synthetic cell culture medium with bovine serum, glutamine, and antibiotics, contributes to these benefits. We hypothesized that (1) DPR using MEM as a dialysate would increase mesenteric perfusion, improve intestinal mucosal injury, and limit intestinal and hepatic inflammation after intestinal I/R and (2) these improvements would be dependent on endothelial nitric oxide pathways.
Eight-week-old C57Bl6J wild-type (WT) and eNOS Knock Out (eNOS KO) male mice were anesthetized and intestinal ischemia was induced for 60 min. After ischemia, 1 mL of phosphate buffered saline vehicle or MEM was injected into the abdominal cavity. Intestinal perfusion was reassessed after 48 h. Animals were then euthanized, and intestines and livers explanted for histologic and molecular analyses.
DPR with MEM significantly improved mesenteric perfusion compared with vehicle (phosphate buffered saline) as measured by Laser Doppler Imaging (WT + MEM 91.58 ± 13.74%, WT + Vehicle 44.27 ± 11.93%, P < 0.05); however, these benefits were lost when endothelial nitric oxide signaling pathways were ablated (eNOS KO + MEM 21.72 ± 5.67 %, eNOS KO + Vehicle 45.24± 11.31%). WT mice treated with MEM also had significantly better preservation of their mucosal architecture (WT + MEM Mdn = 1.0, interquartile range [IQR] = 1.25, WT + Vehicle Mdn = 3.0, IQR = 2.0, P < 0.05). When we compared eNOS KO mice treated with either MEM or vehicle the protective effect of MEM disappeared (eNOS KO + MEM Mdn = 2.0, IQR = 2.25, eNOS KO + Vehicle Mdn = 2.0, IQR = 1.0 P > 0.05). Intestinal levels of interleukin (IL)-1β were increased in WT animals treated with MEM compared with eNOS KOs, whereas concentrations of intestinal IL-6 were similar between groups. Hepatic levels of both IL-1β and IL-6 were significantly elevated in eNOS KOs compared with WT treated with MEM.
DPR with MEM has significant therapeutic potential for improving mesenteric perfusion, intestinal injury, and the local inflammatory response after intestinal I/R. These benefits appear to be dependent on nitric oxide signaling within the endothelium.
直接腹膜复苏（DPR）已被证明可提高肠缺血再灌注损伤（I/R）后的生存率。我们之前已经认识到，最低限度基本培养基（MEM），一种含有牛血清、谷氨酰胺和抗生素的合成细胞培养基，有助于这些益处。我们假设：（1）使用MEM作为透析液的DPR将增加肠系膜灌注，改善肠黏膜损伤，并限制肠I/R后的肠道和肝脏炎症；（2）这些改善将依赖于内皮型一氧化氮途径。
将8周龄的C57Bl6J野生型（WT）和内皮型一氧化氮合酶基因敲除（eNOS KO）雄性小鼠麻醉，诱导肠缺血60分钟。缺血后，向腹腔内注射1mL磷酸盐缓冲盐水赋形剂或MEM。48小时后重新评估肠灌注。然后对动物实施安乐死，取出肠道和肝脏进行组织学和分子分析。
通过激光多普勒成像测量，与赋形剂（磷酸盐缓冲盐水）相比，MEM进行的DPR显著改善了肠系膜灌注（WT + MEM为91.58±13.74%，WT + 赋形剂为44.27±11.93%，P < 0.05）；然而，当内皮型一氧化氮信号通路被消除时，这些益处消失了（eNOS KO + MEM为21.72±5.67%，eNOS KO + 赋形剂为45.24±11.31%）。用MEM治疗的WT小鼠的黏膜结构也得到了显著更好的保存（WT + MEM中位数 = 1.0，四分位间距[IQR] = 1.25，WT + 赋形剂中位数 = 3.0，IQR = 2.0，P < 0.05）。当我们比较用MEM或赋形剂治疗的eNOS KO小鼠时，MEM的保护作用消失了（eNOS KO + MEM中位数 = 2.0，IQR = 2.25，eNOS KO + 赋形剂中位数 = 2.0，IQR = 1.0，P > 0.05）。与eNOS KO小鼠相比，用MEM治疗的WT动物的肠道白细胞介素（IL）-1β水平升高，而各组之间肠道IL-6的浓度相似。与用MEM治疗的WT小鼠相比，eNOS KO小鼠肝脏中的IL-1β和IL-6水平均显著升高。
MEM进行的DPR在改善肠I/R后的肠系膜灌注、肠损伤和局部炎症反应方面具有显著的治疗潜力。这些益处似乎依赖于内皮细胞内的一氧化氮信号传导。
