Lee S W, Feingold D L, Carter J J, Zhai C, Stapleton G, Gleason N, Whelan R L
Division of Colon and Rectal Surgery, New York Presbyterian Hospital, Cornell-Weill University, New York, NY 10021, USA.
Surg Endosc. 2003 Dec;17(12):1996-2002. doi: 10.1007/s00464-003-8154-5. Epub 2003 Oct 23.
It has been well established that open abdominal surgery results in systemic immunosuppression postoperatively; in contrast, laparoscopic surgery is associated with significantly better preserved systemic immune function. However, when intraperitoneal (local) immune function is considered, laparoscopic procedures done under a CO2 pneumoperitoneum (pneumo) have been shown to result in greater immunosuppression compared to that of open surgery. Few studies have simultaneously assessed systemic and local immune function. The purpose of this study was to assess peripheral blood mononuclear cell (PBMC) and peritoneal macrophage tumor necrosis factor-alpha (TNF-alpha) levels, H2O2 production, and MHC class II antigen expression after open and laparoscopically assisted cecectomy in a rat model.
A total of 75 Sprague Dawley rats were used for three separate experiments. For each study, rats were randomly divided into three groups: anesthesia alone (AC), laparoscopic-assisted cecectomy (LC), and open cecectomy via full laparotomy (OP). A CO2 pneumo was used for laparoscopic operations. On postoperative day 1 the animals were sacrificed, macrophages were harvested via intraperitoneal lavage, and PBMCs were isolated from whole blood obtained by cardiac puncture. In experiment 1, macrophages and PBMC from each animal were stimulated with lipopolysaccharide, after which TNF-alpha levels of the supernatant were determined. In experiment 2, after stimulation with PMA, H2O2 release was assessed by measuring fluorescence. In experiment 3, via flow cytometry, the number of cells with surface MHC class II proteins were determined. Data from the three groups in each experiment were compared using analysis of variance Tukey-Kramer tests.
Macrophages and PBMC from rats in the OP group released significantly more TNF-alpha than cells from rats in the LC ( p < 0.05) or AC ( p < 0.05) groups. Macrophages from rats in the OP group released significantly less H2O2 than cells from the AC ( p < 0.01) and LC ( p < 0.05) groups. There was no difference between the AC and LC results. No significant differences in PBMC H2O2 release were noted among any of the groups. OP group macrophages expressed significantly less MHC class II antigen than did AC group macrophages ( p < 0.05). No differences were noted among the LC results and either the OP or AC group's outcomes. No differences were noted in PBMC MHC class II expression among any of the groups.
In all instances, the LC group's macrophage results were similar to the AC group's results. OC group macrophages produced significantly more TNF-alpha and less H2O2 than both the AC and LC groups. MHC class II protein expression was less for the OC group than for the AC group. OC group PBMCs produced more TNF-alpha. No differences in PBMC H2O2 release or MHC class II expression were noted. Laparoscopic methods better preserves the baseline values of the parameters studied.
众所周知，开腹手术术后会导致全身免疫抑制；相比之下，腹腔镜手术与更好地保留全身免疫功能相关。然而，当考虑腹腔内（局部）免疫功能时，与开腹手术相比，在二氧化碳气腹（气腹）下进行的腹腔镜手术已被证明会导致更大程度的免疫抑制。很少有研究同时评估全身和局部免疫功能。本研究的目的是评估大鼠模型中开腹和腹腔镜辅助盲肠切除术后外周血单核细胞（PBMC）和腹腔巨噬细胞肿瘤坏死因子-α（TNF-α）水平、H2O2产生以及MHC II类抗原表达。
总共75只Sprague Dawley大鼠用于三个独立实验。对于每项研究，大鼠被随机分为三组：单纯麻醉组（AC）、腹腔镜辅助盲肠切除组（LC）和经全腹剖腹的开腹盲肠切除组（OP）。腹腔镜手术使用二氧化碳气腹。术后第1天处死动物，通过腹腔灌洗收集巨噬细胞，并从心脏穿刺获得的全血中分离PBMC。在实验1中，用脂多糖刺激每只动物的巨噬细胞和PBMC，然后测定上清液中的TNF-α水平。在实验2中，用佛波酯刺激后，通过测量荧光评估H2O2释放。在实验3中，通过流式细胞术测定表面有MHC II类蛋白的细胞数量。每个实验中三组的数据使用方差分析Tukey-Kramer检验进行比较。
OP组大鼠的巨噬细胞和PBMC释放的TNF-α明显多于LC组（p < 0.05）或AC组（p < 0.05）大鼠的细胞。OP组大鼠的巨噬细胞释放的H2O2明显少于AC组（p < 0.01）和LC组（p < 0.05）的细胞。AC组和LC组的结果无差异。任何组之间PBMC的H2O2释放均无显著差异。OP组巨噬细胞表达的MHC II类抗原明显少于AC组巨噬细胞（p < 0.05）。LC组的结果与OP组或AC组的结果之间无差异。任何组之间PBMC的MHC II类表达均无差异。
在所有情况下，LC组的巨噬细胞结果与AC组的结果相似。OC组巨噬细胞产生的TNF-α明显多于AC组和LC组，而H2O2则少于这两组。OC组的MHC II类蛋白表达低于AC组。OC组PBMC产生更多的TNF-α。PBMC的H2O2释放或MHC II类表达无差异。腹腔镜方法能更好地保留所研究参数的基线值。
