七氟烷体外暴露对乳腺癌患者外周血CD4^+T淋巴细胞亚群表达的影响

Effects of sevoflurane on the expression subsets of CD4+ T lymphocytes in the peripheral blood of breast cancer patients in vitro

目的研究七氟烷在体外对乳腺癌患者外周血CD4^+T淋巴细胞亚群表达的影响。方法采集60例乳腺癌患者外周静脉血,采用免疫磁珠分离出CD4^+T淋巴细胞、Th1细胞和调节性T细胞(Treg细胞),均分为七氟烷实验组(S组)和空白对照组(C组);S组细胞暴露于3%七氟烷2h后置于细胞培养箱,C组细胞直接置于细胞培养箱中。两组均加入T细胞生长因子进行细胞培养,培养后24h,用流式细胞仪检测两组CD4^+T淋巴细胞亚群表达情况,用Annexin V-FITC/PI细胞凋亡检测试剂盒检测两组Th1细胞凋亡情况,用羟基荧光素二醋酸盐琥珀酰亚胺脂(CFSE)细胞增殖检测试剂盒检测两组Treg细胞增殖情况。结果流式细胞仪检测结果显示S组CD4^+T淋巴细胞亚群中Th1细胞的表达较C组下降(P<0.001),而Treg细胞的表达较C组上升(P<0.001);凋亡检测结果显示七氟烷暴露后Th1细胞的凋亡较C组显著增加(P<0.01);CFSE检测结果显示七氟烷暴露后Treg细胞的增殖较C组显著增加(P<0.001)。结论七氟烷处理体外乳腺癌患者外周血CD4^+T淋巴细胞,可导致Th1细胞表达下降,Treg细胞表达增加,其机制可能与促进Th1细胞的凋亡及Treg细胞的增殖有关。

Objective To investigate the effects of sevoflurane on expression subsets of CD4^+T lymphocytes in the peripheral blood of breast cancer patients in vitro.Methods The peripheral blood was collected from 60 breast cancer patients,and then,the CD4^+T lymphocytes,Th1 cells,and Treg cells were sorted by immunomagnetic beads.All cells were divided into sevoflurane group(group S)and control group(group C).Group S was placed in a cell culture incubator after exposing to 3%sevoflurane for 2 h,and group C was placed directly in the cell culture incubator.T cell growth factors were added to both groups.We used flow cytometry to detect the expression of CD4^+T lymphocyte subsets in both groups after culturing for 24 h.The apoptosis of Th1 cells was detected by Annexin V-FITC/PI apoptosis assay kit and the proliferation of Treg cells was detected by 5,6-carboxyfluorescein diacetate,succinimidyl ester(CFSE)cell proliferation assay kit.Results Flow cytometry results showed that the expression of Th1 cells in CD4^+T lymphocyte in group S was lower than that in group C(P<0.001),while the expression of Treg cells was higher(P<0.001).The apoptosis of Th1 cells was significantly increased after exposure to sevoflurane(P<0.01).The results of CFSE showed that the proliferation of Treg cells in group S was significantly higher than that in group C(P<0.001).Conclusions The exposure of peripheral blood CD4^+T lymphocytes of breast cancer patients to sevoflurane in vitro can lead to a reduction in the expression of Th1 cells and an increase in the expression of Treg cells.The mechanism may be related to the promotion of apoptosis of Th1 cells and the proliferation of Treg cells.