医源性细胞外基质破坏作为术后肠梗阻的局部触发因素。
Iatrogenic extracellular matrix disruption as a local trigger for postoperative ileus.
机构信息
Department of General, Visceral, Thoracic, and Vascular Surgery, University of Bonn, Bonn, Germany.
出版信息
J Surg Res. 2012 Dec;178(2):632-9. doi: 10.1016/j.jss.2012.05.044. Epub 2012 Jun 1.
BACKGROUND
Active matrix metallopeptidase 9 (MMP-9) disruption of the extracellular matrix (ECM) plays an important role in inflammatory disorders. In this study, we investigated the inflammatory role of MMP-9 and the ECM breakdown product hyaluronan as a trigger for the postoperative intestinal inflammatory response of postoperative ileus.
METHODS
We performed a standardized intestinal surgical manipulation on rats to produce ileus assessed by the oral non-digestible fluorescein isothiocyanate-dextran transit assay. We studied isolated intestinal muscularis extracts for mRNA expressions of interleukin 6 (IL-6), MMP-9 and CD44. We quantified peritoneal MMP-9 activity using zymography, and quantified peritoneal fluid and serum for hyaluronan and tissue inhibitor of metalloproteinase 1 levels by enzyme-linked immunosorbent assay (ELISA). We cultured peritoneal macrophages and exposed them to peritoneal fluid or synthetic hyaluronan for ELISA analysis of IL-6 and macrophage inflammatory protein-1α.
RESULTS
Transit was significantly delayed after surgical manipulation, and extracts of the isolated jejunal and colonic muscularis demonstrated a significant induction of IL-6, MMP-9, and CD44 mRNAs compared with controls. Zymography confirmed significant MMP-9 activity in peritoneal fluid compared with controls. Enzyme-linked immunosorbent assay measurements showed a significant up-regulation in hyaluronan and tissue inhibitor of metalloproteinase 1 in the peritoneal fluid and serum. In addition, ELISA and reverse transcriptase-polymerase chain reaction measurements of peritoneal macrophages stimulated with postsurgical peritoneal fluid and synthetic hyaluronan resulted in higher expressions of IL-6 and macrophage inflammatory protein-1α in the macrophage supernatant.
CONCLUSIONS
Our results confirm that MMP-9 disruption in the ECM with hyaluronan release and muscularis CD44 receptor induction has the potential to trigger muscularis proinflammatory cascades that cause postoperative ileus. Matrix metallopeptidase 9 inhibition may be a novel therapeutic approach to limit postoperative ileus.
背景
活性基质金属蛋白酶 9（MMP-9）对细胞外基质（ECM）的破坏在炎症性疾病中起着重要作用。在这项研究中，我们研究了 MMP-9 的炎症作用以及 ECM 分解产物透明质酸作为术后肠梗阻术后肠道炎症反应触发因素的作用。
方法
我们对大鼠进行了标准化的肠道手术操作，通过口服不可消化的荧光素异硫氰酸酯-葡聚糖转运试验评估肠梗阻。我们研究了分离的肠肌提取物中白细胞介素 6（IL-6）、MMP-9 和 CD44 的 mRNA 表达。我们使用酶谱法定量腹膜 MMP-9 活性，并通过酶联免疫吸附试验（ELISA）定量腹膜液和血清中的透明质酸和金属蛋白酶组织抑制剂 1 水平。我们培养了腹膜巨噬细胞，并将其暴露于腹膜液或合成透明质酸中，通过 ELISA 分析 IL-6 和巨噬细胞炎症蛋白-1α。
结果
手术后，转运明显延迟，与对照组相比，分离的空肠和结肠肌提取物中 IL-6、MMP-9 和 CD44 mRNA 的诱导明显增加。酶谱法证实腹膜液中 MMP-9 活性明显增加。酶联免疫吸附试验测量显示，腹膜液和血清中的透明质酸和金属蛋白酶组织抑制剂 1 水平明显上调。此外，用术后腹膜液和合成透明质酸刺激腹膜巨噬细胞的 ELISA 和逆转录-聚合酶链反应测量结果表明，巨噬细胞上清液中 IL-6 和巨噬细胞炎症蛋白-1α的表达更高。
结论
我们的结果证实，ECM 中 MMP-9 的破坏与透明质酸的释放和肌层 CD44 受体的诱导有可能引发导致术后肠梗阻的肌层促炎级联反应。基质金属蛋白酶 9 抑制可能是限制术后肠梗阻的一种新的治疗方法。
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