羧甲基茯苓多糖抑制人外周血源性树突状细胞凋亡的体外研究

目的 :探讨羧甲基茯苓多糖(carboxymethytl pachymaram,CMP)对人外周血源性树突状细胞(dendritic cells,DC)凋亡的影响。方法:人外周血单核细胞经rh GM-CSF和IL-4诱导分化为不成熟DC。LPS诱导成熟的基础上分别加入不同浓度的CMP(终浓度为10、50、100 mg/L),以不加CMP为对照组,应用流式细胞术检测细胞凋亡率和趋化因子受体CCR7表达情况,Q-PCR检测CCR7基因表达情况,Western blot检测磷酸化Akt、Akt蛋白表达情况。结果:经CMP处理后DC凋亡率低于对照组,而CCR7的表达高于对照组,磷酸化Akt蛋白表达亦增高,且呈剂量依赖趋势。100 mg/L CMP处理组细胞凋亡率显著低于对照组,差异有统计学意义(P<0.01)。50 mg/L和100 mg/L CMP处理组CCR7的表达和磷酸化Akt蛋白均显著高于对照组,差异均有统计学意义(P<0.05)。结论:CMP能抑制成熟DC的凋亡,其机制可能是通过上调CCR7的表达,经PI3K/Akt信号通路发挥作用。

Carboxymethytl pachymaram inhibits apoptosis of human monocyte-derived dendritic cells

Objective:To investigate the effect of carboxymethytl pachymaram(CMP) on inhibiting apoptosis of human monocyte-derived dendritic cells in vitro. Methods:Human dendritic cells were induced from the peripheral blood monocytes in vitro using recombined human GM-CSF and interleukin (IL)-4,and culture in the presence of CMP at different concentrations (10,50,and 100 mg/L). The percentage of apoptosis and phosphorylated Akt proteins in mature DCs was measured in the presence or absence of CMP by flow cytometry and Western blot,respectively;The expression of CCR7 in mature DCs was measured in the presence or absence of CMP by flow cytometry and Q-PCR,respectively. Results:In CMP-induced DC, the percentage of apoptosis was significantly lower(in the presence of 100mg/L CMP) and the expression of CCR7 and phosphorylated Akt protein significantly higher(in the presence of 50mg/L and 100mg/L CMP) than that of control group,both in a dose-dependent model. Conclusion:CMP can inhibit the apoptosis of human monocyte-derived dendritic cells and its anti-apoptotic mechanism is probably mediated by CCR7 and PI3K/Akt signaling pathway.