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探讨辣椒素对人γδT细胞体外增殖及杀伤骨肉瘤细胞的影响及作用机制。分离健康人PBMC,加入到含异戊烯焦磷酸(isopentenyl pyrophosphate,IPP)和IL-2的DMEM-F12完全培养基中诱导培养γδT细胞。不同浓度的辣椒素作用于γδT细胞48h后,CCK-8法检测各药物浓度组γδT细胞的增殖能力,采用流式细胞术检测各组γδT细胞穿孔素、颗粒酶B、CD107a及IFN-γ的表达,用LDH释放法检测各组γδT细胞对骨肉瘤HOS细胞的体外杀伤活性。结果显示,培养7d时各组γδT细胞纯度达到了(85.33±6.07)%。浓度为3.2~50μmol/L的辣椒素能显著促进γδT细胞的增殖,在浓度为25μmol/L时其穿孔素、颗粒酶B、CD107a及IFN-γ的表达均显著高于对照组,且对人骨肉瘤HOS细胞的体外杀伤活性显著增强。以上结果提示辣椒素在体外能通过促进γδT细胞穿孔素、颗粒酶B和IFN-γ的表达上调显著增强其抗骨肉瘤细胞活性。
To investigate the effect of capsaicin on human γδT cells in vitro proliferation and kill osteosarcoma cells and its mechanism. Healthy PBMCs were isolated and cultured in DMEM-F12 complete medium containing isopentenyl pyrophosphate (IPP) and IL-2 to induce the induction of γδT cells. After treated with different concentrations of capsaicin for 48 h, the proliferation of γδT cells in each drug concentration group was determined by CCK-8 assay. The perforin, granzyme B, CD107a and IFN-γ in each group were detected by flow cytometry The cytotoxicity of γδT cells to HOS cells in vitro was detected by LDH release assay. The results showed that the purity of γδT cells in each group reached (85.33 ± 6.07)% at 7 days. The concentration of 3.2 ~ 50μmol / L capsaicin can significantly promote the proliferation of γδT cells, the concentration of 25μmol / L perforin, granzyme B, CD107a and IFN-γ were significantly higher than the control group, and the human bone Sarcoma HOS cells in vitro cytotoxic activity was significantly enhanced. The above results suggest that capsaicin can significantly enhance the anti-osteosarcoma cell activity in vitro by up-regulating the expression of perforin, granzyme B and IFN-γ in γδT cells.