不同结构的中/长链脂肪乳对肿瘤病人免疫细胞的影响

Impact of different structure medium-and long-chain triglyceride emulsions on immune cells in gastrointestinal tumor patients

目的:探讨不同结构的中/长链脂肪乳对胃肠肿瘤病人免疫细胞功能的影响。方法:①将胃肠肿瘤病人的外周血分别与不同剂量(10mg/mL、20mg/mL、100mg/mL)的结构脂肪乳(STG)、物理混合的中/长链脂肪乳(MCT/LCT)在体外共同孵育2h后,以FITC-葡聚糖(dextran)为吞噬标志,应用流式细胞技术检测中性粒细胞的吞噬率;②将分离的淋巴细胞分别在含不同剂量(10mg/mL、20mg/mL、100mg/mL)的结构脂肪乳或物理混合的中/长链脂肪乳培养基中孵育24h,应用Annexin-V/PI双染色方法测定淋巴细胞的凋亡率。结果:①在含10mg/mL脂肪乳的培养环境中,STG组、MCT/LCT组和空白对照组的中性粒细胞吞噬率间差异无统计学意义;②在含20mg/mL脂肪乳的培养环境中,STG组的中性粒细胞吞噬率显著低于MCT/LCT组(P=0.024),但其与空白对照组间差异无统计学意义;③在含100mg/mL脂肪乳的培养环境中,STG组的中性粒细胞吞噬率均显著低于MCT/LCT组和空白组(P<0.05);④随着脂肪乳浓度的增加,STG组的中性粒细胞吞噬率呈下降趋势(P<0.05);⑤STG组和MCT/LCT组的淋巴细胞凋亡率不随培养环境中脂肪乳浓度增加而改变,且2组间差异无统计学意义。结论:模拟体内常用的脂肪乳剂量时,结构脂肪乳不明显改变胃肠肿瘤病人的中性粒细胞吞噬功能和淋巴细胞凋亡率。大剂量应用脂肪乳时,结构脂肪乳能抑制中性粒细胞的吞噬功能,而物理混合的中/长链脂肪乳则无此现象。

Objective To study the influence of different structure medium-and long-chain triglyceride emulsions on immune cells functions in gastrointestinal tumor patients. Methods ① Human whole blood cells were cultured in vitro with the structured triglyceride （STG） emulsion or physical mixture of medium- and long-chain triglyceride （MCT/ LCT） emulsion with a final concentration of 10 mg/mL, 20 mg/mL and 100 mg/mL respectively. Taking the FITC-dextran particle as a marker of phagocytosis, the percentage of neutrophil phagoeytosis was detected by flow eytometry after 2 hours of the incubation. ② The isolated lymphocytes were incubated in the STG emulsion or MCT/LCT emulsion with a final concentration of 10 mg/mL, 20 mg/mL and 100 mg/mL respectively. After 24 hours of incubation, the apoptosis rate of lymphocytes was measured by Annexin V/PI staining with flow cytometry. Results ①With lipid emulsions at the concentration of 10 mg/mL, there was no difference in neutrophil phagocytosis rate among the STG, MCT/LCT and the blank groups. ②At the concentration of 20 mg/mL, the neutrophil phagocytosis rate in the STG group was significant lower than that in the MCT/LCT group（P〈0.05 ）, but there was no significant difference between the STG and the blank groups.③） At the concentration of 100 mg/mL, the neutrophil phagocytosis rate in the STG group was significant lower than that in the MCT/LCT and blank groups （P〈0.05）. ④） The tendency of neutrophil phagocytosis decreased following the upregulation of lipid emulsion dosage（P〈0.05）.⑤After 24-hour cell culture, there was no obvious change in the apoptosis rate of lymphocytes with the changes in concentration of lipid emulsions in the STG or MCT/LCT groups. Conclusions There is no significant change in neutrophil phagocytosis function and lymphocyte apoptosis with structured triglyceride emulsion at a regular dosage. However, neutrophil phagoeytosis function is inhibited with structured triglyceride emulsion at a high dosage. No such phenomena is observed when using the MCT/LCT emulsion.