细胞外组蛋白诱导小鼠原代单核细胞死亡的实验研究

The study on the induction of mouse primary monocyte death by extracellular histone

为探讨细胞外组蛋白对小鼠原代单核细胞活力的影响以及可能机制,将小鼠原代单核细胞培养于含不同浓度组蛋白的培养基上,采用MTT染色法检测细胞活性并筛选合适的组蛋白浓度。将原代单核细胞培养于特定浓度组蛋白培养基上,利用MTT染色法检测不同时间的细胞活性。分别用自噬抑制剂和死亡抑制剂预处理细胞,采用MTT染色法结合PI-Annexin V双染法、流式细胞术检测细胞活性和死亡率。与对照组比较,组蛋白组细胞死亡率明显升高(P < 0.05),应用死亡抑制剂后,细胞死亡率明显下降(P < 0.05)。细胞外组蛋白可抑制原代单核细胞活性,诱导原代单核细胞死亡,且具有浓度和时间依赖性。自噬可能会增加组蛋白诱导的单核细胞死亡。

To investigate the effect of extracellular histone on the viability of mouse primary monocytes and its possible mechanism, mouse primary monocytes were cultured in mediums containing different concentrations of histone. The cell viability was detected and the appropriate histone concentration was determined screened by MTT staining. Primary monocytes were cultured in a specific concentration of histone medium, and the cell viability at different culture times was measured by MTT staining. In addition, cells were pretreated with autophagy inhibitor and cell death inhibitor, respectively. And the cell viability and mortality were detected by MTT staining combined with PI-Annexin V double staining and flow cytometry. Compared with the control group, the cell death rate of the histone group was significantly increased (P < 0.05), and the cell death rate was significantly decreased after the cell death inhibitor was used (P < 0.05). Taken together, our study indicates extracellular histone could inhibit primary monocyte viability and induce death of primary monocytes in a concentration- and time-dependent manner. Autophagy could increase histone-induced monocyte death.