CAJ | 학술논문

目的：探讨槲皮苷（ quercitrin ）对人外周血γδT细胞体外增殖和功能影响及相关机制。方法从健康人外周血中分离单个核细胞（PBMC），加入到含有异戊烯焦磷酸和IL-2的RPMI 1640完全培养基中诱导培养获得γδT细胞。用不同浓度的槲皮苷作用γδT细胞48 h后，用CCK-8法测定不同浓度槲皮苷组γδT细胞增殖和杀伤能力；采用流式细胞术检测各组γδT细胞颗粒酶B和穿孔素的表达；用Western blot法检测p-ERK、p-Akt和Bcl-2蛋白的表达。结果培养10 d后的γδT细胞纯度达到（88．94±2．36）％。槲皮苷浓度在10～80μg/ml时能显著促进γδT细胞的增殖及颗粒酶B、穿孔素的表达和对结肠癌HCT116细胞的杀伤能力。在蛋白水平p-ERK、p-Akt和Bcl-2的表达量均显著高于对照组，而内参GAPDH表达量无变化。结论槲皮苷可体外增强γδT细胞的增殖及杀伤功能，其机制可能通过p-ERK和p-Akt信号通路。
목적：탐토곡피감（ quercitrin ）대인외주혈γδT세포체외증식화공능영향급상관궤제。방법종건강인외주혈중분리단개핵세포（PBMC），가입도함유이무희초린산화IL-2적RPMI 1640완전배양기중유도배양획득γδT세포。용불동농도적곡피감작용γδT세포48 h후，용CCK-8법측정불동농도곡피감조γδT세포증식화살상능력；채용류식세포술검측각조γδT세포과립매B화천공소적표체；용Western blot법검측p-ERK、p-Akt화Bcl-2단백적표체。결과배양10 d후적γδT세포순도체도（88．94±2．36）％。곡피감농도재10～80μg/ml시능현저촉진γδT세포적증식급과립매B、천공소적표체화대결장암HCT116세포적살상능력。재단백수평p-ERK、p-Akt화Bcl-2적표체량균현저고우대조조，이내삼GAPDH표체량무변화。결론곡피감가체외증강γδT세포적증식급살상공능，기궤제가능통과p-ERK화p-Akt신호통로。
Objective To investigate the in vitro effects of quercitrin on the proliferation and the cytotoxicity of human γδT cells.Methods Peripheral blood mononuclear cells (PBMCs) were isolated from healthy subjects and cultured with isopentenyl pyrophosphate and IL -2 to induce human γδT cells.The hu-manγδT cells were cultured with quercitrin at various concentrations for 48 hours.CCK-8 kits were used to analyze the in vitro proliferation and cytotoxic activities of γδT cells.Flow cytometry was performed to meas-ure the expression of granzyme B and perforin in γδT cells.The expression of p-ERK, p-Akt and Bcl-2 at protein level were detected by Western blot .Results The percentage of human γδT cells in PBMCs was in-creased from (2.96±1.83)%to (88.94±2.36)%after 10 days of culture.The quercitrin at concentrations of 10 to 80 μg/ml could promote the growth of γδT cells and up-regulate the expression of granzyme B , per-forin, p-ERK, p-Akt and Bcl-2 in a dose dependent manner .The cytolytic activities of γδT cells against co-lonic carcinoma cells ( HCT116 ) were enhanced by quercitrin .Conclusion Quercitrin could promote the proliferation of γδT cells and enhance the expression of granzyme B and perforin at certain concentrations in vitro.ERK1/2 and Akt signal transduction systems might be involved in the process .