模拟CO2气腹对腹膜间皮细胞功能及其对胃癌细胞黏附的影响

Effects of simulated CO2 pneumoperitoneum upon function of peritoneal mesothelial cells and adherence to gastric cancer cells

目的通过体外模拟CO2气腹环境，观察其对腹膜间皮细胞损伤、增殖、分泌功能及对人胃腺癌细胞AGS、MKN-45与间皮细胞黏附的影响。方法以体外原代培养腹膜间皮细胞为实验对象。实验分为对照组和CO2处理组，其中CO2处理组又按照压力不同分为10、12、15mmHg组。间皮细胞经不同压力CO2处理后用乳酸脱氢酶法检测细胞损伤，氯化硝基四氦唑蓝法检测细胞增殖活性，免疫组化法检测细胞分泌功能，流式细胞仪检测胃腺癌细胞与间皮细胞黏附率。结果10、12mmHg组LDH水平与对照组相比差异无统计学意义（P〉0．05），15mmHg组较对照组高（122．79±4．28比100．89±1．22，P〈0．05）。10、12mmHg组间皮细胞增殖活性与对照组比较差异无统计学意义（P〉0．05）；15minHg组前48h内与对照组比较差异无统计学意义（P〉0．05），72h时较对照组低（1．08±0．05比1．21±0．05，P〈0．05）。各CO2气腹组白细胞介素6（TL-6）表达水平较对照组高（P〈0．05），且15mmHg组IL-6表达水平持续升高（P〈0．05）。各CO2气腹组AGS、MKN45细胞与腹膜间皮细胞黏附率与对照组比较差异均无统计学意义（均P〉0．05）。结论模拟10、12mmHgCO2气腹环境对腹膜间皮细胞的损伤、增殖活性无明显影响，而15mmHgCO2气腹环境抑制腹膜间皮细胞增殖并造成间皮细胞损伤。CO2气腹环境使间皮细胞IL-6表达水平升高，可能有利于腹膜间皮细胞发挥免疫功能。CO2气腹环境并不促进胃癌细胞与腹膜间皮细胞黏附。

Objective To investigate the effects of CO2 insufflation with different pressures upon the injury, proliferation and secretion of peritoneal mesothelial cells and the possible effects of gastric cancer cells adhering to mesothelial cells. Methods Primary cultured peritoneal mesothelial cells were divided into CO2 and control group. In CO2 group, the cells were exposed to CO2 at different pressures （10, 12 and 15 mm Hg）. And then the cellular injury was detected by LDH assay, the proliferation quantified by MTF assay and the secretion measured by ELISA. Flow cytometry was used to analyze the adherent rate between gastric cancer cells and mesothelial cells. Results The LDH assay showed that 10 and 12 mm Hg subgroups were not significantly different from that in the control group （ P 〉 0.05 ） whereas the 15 mm Hg subgroup was higher than the control group（ 122.79 ±4.28 vs 100.89± 1.22, P 〈0.05）. MTT assay indicated that 10 and 12 mm Hg subgroups were similar to that of the control group （ P 〉 0.05）. However the 15 mm Hg subgroup decreased to 1.08±0.05 at 72 h post-experiment. But the same finding was not observed during first 48 h. The concentration of IL-6 in each group increased as compared with the control group （ 1.21 ± 0. 05, P 〈 0.05）. Moreover this tendency was persistent only in the 15 mm Hg subgroup （P 〈 0.05）. The results of flow cytometry showed that the adherent rate between gastric cancer cells and mesothelial cells in each CO2 group were similar to the control group （all P 〉 0.05 ）. Conclusion Pressures of CO2 at 10 aud 12 mm Hg have no effect upon the injury and proliferation of peritoneal mesothelial cells. When pressure rises to 15 mm Hg, the cellular injury becomes apparent and the proliferation is inhibited. The CO2 environment up-regulates the level of IL-6. It may benefit to the local immune reaction of peritoneum. The CO2 pneumoperitoneum does not promote the adhesion of gastric cancer cells to mesothelial cells.