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目的检测牙龈卟啉单胞菌脂多糖(LPS)对人外周血中T淋巴细胞活化及凋亡的作用。方法选取10名全身及牙周组织健康受试者,分离外周血中T淋巴细胞,在有/无LPS情况下培养0~96h,用荧光探针(Annexin VFITC、PI、CD69)进行标记,并进行流式细胞仪检测。结果 T淋巴细胞+LPS组Annexin V+/PI-细胞百分数在48h、72h、96h分别为16.36±2.22,27.16±2.81,47.17±4.70。T淋巴细胞组AnnexinV+/PI-细胞百分数在48h、72h、96h分别为7.55±2.76,6.32±1.48,12.03±3.24,两组间存在明显差异(P<0.01)。CD69+T淋巴细胞+LPS组和CD69+T淋巴细胞组AnnexinV+/PI-细胞百分数除24 h外的4个时间点上都无明显差异(P>0.05)。CD69+淋巴细胞+LPS组Annexin V+/PI-细胞百分数在各个时间点上都明显高于T淋巴细胞+LPS组(P<0.01)。结论 LPS能够诱导人外周血中T淋巴细胞活化,并且能够通过活化促进其凋亡。
Objective To detect the effect of Porphyromonas gingivalis lipopolysaccharide (LPS) on the activation and apoptosis of T lymphocytes in human peripheral blood. Methods T-lymphocytes were isolated from 10 whole body and periodontal healthy volunteers. Peripheral blood T lymphocytes were isolated and cultured for 0-96h with or without LPS. The cells were labeled with Annexin VFITC (PI, CD69) Flow cytometry. Results The percentages of Annexin V + / PI-cells in T lymphocytes + LPS group were 16.36 ± 2.22, 27.16 ± 2.81 and 47.17 ± 4.70 at 48h, 72h and 96h respectively. The percentages of Annexin V / PI-cells in T lymphocyte group were 7.55 ± 2.76, 6.32 ± 1.48 and 12.03 ± 3.24 at 48h, 72h and 96h, respectively. There was significant difference between the two groups (P <0.01). The percentages of Annexin V / PI-cells in CD69 + T lymphocytes + LPS group and CD69 + T lymphocyte group had no significant difference at 4 time points except 24 h (P> 0.05). The percentage of Annexin V + / PI-cells in CD69 + lymphocytes + LPS group was significantly higher than that in T lymphocytes + LPS group at all time points (P <0.01). Conclusion LPS can induce T lymphocyte activation in human peripheral blood and promote its apoptosis through activation.