高原红细胞增多症患者中性粒细胞[Ca^(2+)]_i及NADPH氧化酶活性变化

Changes of neutrophils [Ca^(2+)]_i and activity of NADPH oxidase in patients with high altitude polycythemia

目的观察高原红细胞增多症(HAPC)患者外周血中性粒细胞[Ca2+]i及NADPH氧化酶活性变化。方法 HAPC患者30例(HAPC组),世代居住的健康者29例(健康组),分离两组外周血中性粒细胞,两组细胞各添加fMLP(终浓度为0、0.001、0.01、0.1、1、10μmol/L)及PMA(终浓度为0、10、20、30、40、50μmol/L)后,采用钙离子荧光探针(Fura-2/AM)法检测两组中性粒细胞内[Ca2+]i;ELISA法检测中性粒细胞NADPH氧化酶活性。结果 fMLP终浓度为0.1、1、10μmol/L时,健康组中性粒细胞[Ca2+]i分别为(223.6±34.6)、(260.7±30.0)、(316.6±53.8)nmol/L;HAPC组分别为(203.2±25.6)、(235.5±35.9)、(278.1±47.1)nmol/L,两组比较,P<0.05或<0.01。fMLP终浓度为0.1、1、10μmol/L时,健康组中性粒细胞NADPH氧化酶活性分别为(81.3±16.9)、(97.6±23.7)、(119.5±33.8)pmol/(10-6个细胞·30 min);HAPC组分别为(72.0±16.9)、(83.6±15.3)、(102.9±26.3)pmol/(10-6个细胞·30min),两组比较,P<0.05或<0.01。PMA终浓度为30、40、50μmol/L时,健康组中性粒细胞[Ca2+]i分别为(224.9±33.1)、(326.7±45.3)、(646.9±78.1)nmol/L;HAPC组分别为(205.5±26.3)、(293.9±52.6)、(567.9±103.1)nmol/L,两组比较,P<0.05或<0.01。PMA终浓度为30、40、50μmol/L时,健康组中性粒细胞NADPH氧化酶活性分别为(258.2±25.1)、(327.3±38.5)、(383.1±36.7)pmol/(10-6个细胞·30 min);HAPC组分别为(213.8±36.3)、(291.3±50.5)、(337.2±36.0)pmol/(10-6个细胞·30 min),两组比较,P<0.01。结论高原缺氧降低人外周血中性粒细胞NADPH活性及[Ca2+]i浓度升高幅度。

Objective To observe the changes of peripheral blood neutrophils [ Ca2 ＋] i and the activity of NADPH oxi- dase in patients with high altitude polycythemia （HAPC）. Methods Peripheral blood neutrophils in 30 HAPC patients （HAPC group） and 29 cases of generations living healthy subjects （healthy group） were isolated, cells in the two groups were added fMLP （with final concentrations of 0, 0. 001, 0.01,0.1, 1, 10 μmol/L） and PMA （with final concentrations of 0, 10, 20, 30, 40, 50 μmol/L）. After that, Calcium ion florescence prob （Fura-2/AM） was used to detect the changes of [ Ca2＋ ]i in the neutrophils; NADPH oxidase activity of the neutrophils was detected by ELISA. Results When the final concentrations of fMLP were 0.1, 1 and 10 μmol/L, respectively, neutrophil [ Ca2＋]i were separately （223.6 ± 34.6）, （260.7 ± 30） and （316.6 ± 53.8） nmol/L in the healthy group; while they were （203.2 ± 25.6）, （235.5± 35.9） and （ 278.1 ± 47.1 ） mmol/L in the HAPC group （ P 〈 0.05 or 〈 0.01 ）. When the final concentrations of fMLP were 0.1, 1 and 10 μmol/L, the activities of neutrophil NADPH oxidase were respectively （81.3 ±16.9）, （97.6 ± 23.7 ） and （ 119.5 ± 33.8） pmo]/（ 10 ^- 6 cells per 30 min） in the healthy group, while （72 ±16.9）, （83.6 ± 15.3 ） and （ 102.9 ± 26.3 ） pmol/ （ 10 -6 cells per 30 min） in the HAPC group （P 〈0.05 or 〈0.01 ）. When the final concentrations of PMA were 30, 40 and 50 μmol/L, the neutrophil [Ca2＋ ]i were separately （224.9 ±33.1）, （326.7 ±45.3） and （646.9 ± 78.1） nmol/L in the healthy group; while in the HAPC group they were （205.5 ± 26.3 ）, （293.9 ±52.6） and （567.9 ± 103.1 ） mmol/LL, respec-tively （P 〈 0.05 or 〈 0.01 ）. When the final concentrations of PMA were 30, dO and 50 ~mol/L, the activities of neutrophil NADPH oxidase were respectively （258.2 ±25.1 ）, （ 327.3 ± 38.5 ） and （383.1 ± 36.7 ） pmol/（ 10 ^-6 cells per 30 min） in the healthy group, while in the HAPC group they were （213.8 ± 36.3 ）, （ 291.3 ± 50. 5 ） and （ 337.2± 36） pmol/（ 10 ^- 6 cells per 30 min）, respectively （P 〈0.01 ）. Conclusion The high altitude hypoxia can suppress the activity of NADPH oxi- dase in neutrophils and the concentration of intraceUular calcium ion.