目的

观察门静脉高压症（PH）脾脏巨噬细胞（M_φ）游离Ca^2＋浓度和分布的变化，为深入探讨门静脉高压症脾脏的变化及评价其免疫功能状况提供实验依据。

方法

选取门静脉高压症脾亢患者（均为慢性乙型肝炎患者）的手术切除脾脏（12例）为实验组，外伤性脾破裂患者的手术切除脾脏（4例）为正常对照组。贴壁培养法分离纯化脾脏组织M_φ，使用Ca²⁺荧光指示剂Fluo-3/AM负载M_φ，以激光共焦镜显微镜观察M_φ内Ca²⁺的分布，流式细胞仪检测M_φ内Ca²⁺的浓度变化。

结果

与正常脾脏相比，PH脾脏M_φ内Ca²⁺的分布无明显变化，但可见细胞表面有较多的Ca²⁺荧光染色阳性的伪足样突起；PH脾脏M_φ内Ca²⁺浓度为明显升高（48.75±2.61 vs 39.92±1.76，P＜0.05）。

结论

PH脾脏M_φ内Ca²⁺浓度明显升高，进一步说明PH脾脏M_φ处在活化状态，但PH脾脏M_φCa²⁺浓度变化的具体机制，以及这种变化对PH脾脏免疫功能的影响还需要进一步的研究。

Objective

To investigate the changes in concentration and distribution of Ca²⁺ in splenic macrophage (M_φ) in hypersplenism due to portal hypertension (PH), providing the evidence for discussing the changes and the immune function of PH spleen.

Methods

Twelve patients with hypersplenism due to PH and four patients with traumatic rupture of spleen were regarded as hypersplenia and control group respectively.Using Fluo-3/AM as indicator,the concentration and distribution of Ca²⁺in splenic M_φ were detected by laser confocal microscope and flow cytometry,respectively.

Results

As compared to control group,the concentration of Ca²⁺in PH spleen M_φ were increased significantly (48.75±2.61 vs 39.92±1.76, P＜0.05). There were many pseudopod-like prominency with Fluo-3 fluorescence positive on the surface of PH splenic M_φ.

Conclusions

The concentration of Ca²⁺in PH spleen M_φis increased significantly.It further supports the viewpoint that the PH splenic M_φretained at activated state.But it still needs to be cleared that the mechanism of changes in the concentration and distribution of Ca²⁺and its effects on immune function ofPH spleen.