不同气体条件对腹膜形态影响的实验研究

目的 在体实验研究提示,CO2气腹后,腹膜结构出现改变.本实验研究不同气体条件对腹膜形态的影响,探讨CO2气腹对腹腔肿瘤细胞影响的潜在机制.方法 在体实验:乳猪14头,年龄7～14 d,体重2～4 kg.随机分为两组:CO2气腹组(CO2组,n=7),N2O气腹组(N2O组,n=7).分别行100%CO2及100%N2O气腹,气腹时间为4 h,气腹压力为12 mm Hg.实验结束,采集腹膜标本.离体实验:C57BL/6小鼠10只,年龄4周左右,经0.125%胰酶预处理后,通过腹腔灌洗分离培养小鼠腹膜间皮细胞.离体细胞随机分为三组:对照组(5%CO2组).100%CO2组,8 cm H2O压力100%CO2组.细胞培养至连续单层后,将细胞分别暴露于5%CO2、100%CO2及8cmH2O压力100%CO2中4 h.不同气体条件下腹膜及分离培养的间皮细胞行电镜观察.结果 在体实验腹膜电镜结果提示压力12 mm Hg的100%CO2气腹维持4 h,导致腹膜间皮细胞层破坏,基底膜暴露,仅存细胞骨架;相同条件的100%N2O气腹使间皮细胞间隙增大,部分区域基底膜暴露.离体实验观察100%CO2破坏间皮细胞微绒毛,压力下的100%CO2对问皮细胞影响更加明显.结论 100%CO2使在体腹膜及离体间皮细胞超微结构发生明显改变,因此,小鼠模型中发现的CO2气腹后神经母细胞瘤转移增加可能与间皮细胞屏障削弱有关.

Abstract：

Objective Electron microscopic studies have shown significant morphologic changes of peritoneum after CO2 pneumoperitoneum in vivo. This experiment was to assess the effect of different gas on the morphology of peritoneum and the underlying mechanism of CO2 pneumoperitoneum on tumor cells. Methods In vivo, fourteen piglets (2-4 kilogram in weight, 7-14 days of age) were equally divided into the CO2 group(n = 7) and N2O group(n = 7). 100% CO2 or 100% N2O pneumoperitoneum was infused for 4 hours. Pneumoperitoneum pressure was 12 mmHg. At the end of the experiment, the samples of peritoneum were collected. In vitro, primary murine peritoneal mesothelial cells were36 collected by peritoneal lavage from ten C57BL/6 mice (4 weeks of age) after 0. 125% trypsin pretreatment. Isolated cells were divided into three groups: control group (5% CO2),100% CO2 group and 8 cm H2O pressure &. 100% CO2 group. After monolayers of mesothelial cells were established, cells were cultured with 5%CO2, 100% CO2 and 100% CO2 with 8 cm H2O pressure for 4 hours. Peritoneum and isolated mesothelial cells were examined by scanning electron microscopy. Results Scanning electron microscopy investigation suggested in vivo, 12 mmHg 100% CO2 pneumoperitoneum for 4 hours destroyed mesothelial cells layer of peritoneum, exposing the basal lamina. In contrast, 100% N2O pneumoperitoneum leaded to an increase of intercellular gaps and the basal lamina was exposed in part areas under same pressure and duration. In vitro, 100% CO2 exposition was associated with a significant destruction of the microvilli formation of isolated mesothelial cells. 100% CO2 with 8 cm H2O pressure had more significant impact on mesothelial cells. Conclusions The peritoneal mesothelial cells lose their typical cell morphology when exposed to 100% CO2. Thus, the increased neuroblastoma metastasis observed after CO2 pneumoperitoneum in mice might be related to an impaired mesothelial barrier function.

Experimental study on effects of different gas on peritoneum

Objective Electron microscopic studies have shown significant morphologic changes of peritoneum after CO2 pneumoperitoneum in vivo. This experiment was to assess the effect of different gas on the morphology of peritoneum and the underlying mechanism of CO2 pneumoperitoneum on tumor cells. Methods In vivo, fourteen piglets (2-4 kilogram in weight, 7-14 days of age) were equally divided into the CO2 group(n = 7) and N2O group(n = 7). 100% CO2 or 100% N2O pneumoperitoneum was infused for 4 hours. Pneumoperitoneum pressure was 12 mmHg. At the end of the experiment, the samples of peritoneum were collected. In vitro, primary murine peritoneal mesothelial cells were36 collected by peritoneal lavage from ten C57BL/6 mice (4 weeks of age) after 0. 125% trypsin pretreatment. Isolated cells were divided into three groups: control group (5% CO2),100% CO2 group and 8 cm H2O pressure &. 100% CO2 group. After monolayers of mesothelial cells were established, cells were cultured with 5%CO2, 100% CO2 and 100% CO2 with 8 cm H2O pressure for 4 hours. Peritoneum and isolated mesothelial cells were examined by scanning electron microscopy. Results Scanning electron microscopy investigation suggested in vivo, 12 mmHg 100% CO2 pneumoperitoneum for 4 hours destroyed mesothelial cells layer of peritoneum, exposing the basal lamina. In contrast, 100% N2O pneumoperitoneum leaded to an increase of intercellular gaps and the basal lamina was exposed in part areas under same pressure and duration. In vitro, 100% CO2 exposition was associated with a significant destruction of the microvilli formation of isolated mesothelial cells. 100% CO2 with 8 cm H2O pressure had more significant impact on mesothelial cells. Conclusions The peritoneal mesothelial cells lose their typical cell morphology when exposed to 100% CO2. Thus, the increased neuroblastoma metastasis observed after CO2 pneumoperitoneum in mice might be related to an impaired mesothelial barrier function.