硫化氢对U937细胞吞噬能力的影响及其机制研究

Effect of hydrogen sulfide on the phagocytosis by U937 cells and the mechanism

目的研究硫化氢对U937细胞吞噬能力的影响及其可能的机制。方法传代培养U937细胞,进行佛波酯诱导后,分为三组[对照组、硫氢化钠(NaHS)组、炔丙基甘氨酸(PPG)组]进行酵母菌吞噬实验,吞噬作用时间4小时。另分四组[对照组、NaHS组、吡咯烷二硫代氨基甲酸(PDTC)组、PDTC+NaHS组]进行墨汁颗粒吞噬实验,吞噬作用时间2小时。酵母菌吞噬结束后,进行显微镜下计数每百个U937细胞吞噬的酵母菌的数目(即吞噬指数)以及发生吞噬的细胞数目(即吞噬率)。墨汁颗粒吞噬结束时,进行细胞洗涤、破碎,Elisa测定540nm吸光度值。结果 NaHS显著地增加了U937细胞对酵母菌的吞噬率以及吞噬指数。而PPG显著地抑制了U937细胞的吞噬率以及吞噬指数。墨汁颗粒吞噬实验显示,NaHS组U937细胞的吞噬墨汁颗粒OD值显著高于对照组,而PDTC+NaHS组显著低于NaHS组。结论:NaHS促进了U937细胞的吞噬能力,其机制之一可能是通过促进核因子-κB的表达实现的。

Objective To study the effect of hydrogen sulfide on the phagocytosis by U937 cells and the possible mechanism. Method After induction with phorbol myristoyl acetate, U937 cells were divided into 3 groups, and were treated with sodium hydrogenuid （ NaHS, at the final concentration of 200μM） , DL-propargylglycine （ PPG, at the final concentration of 200μM） or no intervention respectively. Then, yeast cells were added into each group. U937 cells with phagocytic yeast cells were calculated. Also, yeast cells phagocyted by U937 were also calculated. Another experiment was conducted to observe the phagocytosis of India ink particles by U937 cells. U937 cells were divided into 4 groups [ control, NaHS, Ammonium pyrrolidinedithiocarbamate （ PDTC） , PDTC＋NaHS] . Two hours after being co-cultured with India ink particles, U937 cells were washed and disrupted. OD values were read with enzyme-linked immuno sorbent assay. Result NaHS promoted phagocytosis by U937 cells. Both phagocytic rate and phagocytic index were increased. On the contrary, PPG inhibited phagocytic rate and phagocytic index. For the ex-periment with India ink particles, NaHS increased the OD value significantly compared with control, while PDTC decreased the OD value significantly compared with NaHS group. Conclusion NaHS promoted the phagocytosis by U937 cells. Promotion of the expression of nuclear factor-κB may be the mechanism.