高分压氧体外抑制大鼠脾脏淋巴细胞功能及N-乙酰半胱氨酸的预防作用

目的 :观察高分压氧体外暴露对脾淋巴细胞功能的影响及 N-乙酰半胱氨酸 (NAC)的预防作用。 方法 :分离成年大鼠脾脏淋巴细胞 ,制成细胞悬液 ,置于压力舱内以 4 0 0 k Pa氧气 (含 1.5 % CO2 )培养 4 h。 NAC预防组在培养液中加入 30mm ol/ L NAC。用流式细胞术测定淋巴细胞亚型 ,3H- Td R掺入法测定淋巴细胞增殖反应及 EL ISA法测定细胞经刀豆球蛋白(Con A )刺激后分泌的 IL - 2水平。 结果 :经高分压氧处理后 ,脾淋巴细胞 CD3+ 、CD4 + CD3+ 及 CD8+ CD3+ 比例均显著下降(P<0 .0 1,P<0 .0 5 ) ,CD4 + CD3+ / CD8+ CD3+ 比值未有变化 ;淋巴细胞增殖率及经 Con A刺激后的 IL - 2分泌量显著减少(P<0 .0 1,P<0 .0 5 )。培养液中添加 30 mm ol/ L NAC后 ,各指标值的下降程度均明显减轻 (P<0 .0 1,P<0 .0 5 )。 结论 :高分压氧对离体大鼠脾淋巴细胞功能有直接抑制作用 ,NAC能部分预防该抑制作用。

In vitro inhibitory effects of hyperbaric oxygen on function of rat splenic lymphocyte and preventive effect of N-acetylcysteine

Objective: To observe the in vitro effects of hyperbaric oxygen on the function of splenic lymphocyte and the preventive effect of N acetylcysteine(NAC). Methods: Splenic lymphocytes of adult rats were separated and cultured under 400 kPa O 2 (containing 1.5% CO 2) in a temperature controlled hyperbaric chamber for 4 h. The culture medium contained 30 mmol/L NAC in NAC preventive group. Then the subpopulation of lymphocytes in spleen were identified using 3 coloured flow cytometry. 3H TdR incorporation were used to assess the proliferation rate of the cells. The cells excretion of IL 2 after ConA stimulation was determined with ELISA. Results: After hyperbaria exposure,CD3 +, CD4 +CD3 + and CD8 +CD3 + cells all decreased( P <0.01, P <0.05) and the CD4 +CD3 +/CD8 +CD3 + ratio had no change. The cell proliferation rate and the IL 2 excretion were all reduced( P <0.01, P <0.05). NAC(30 mmol/L) in culture reduced the decrease of all the above indexes significantly( P <0.01, P <0.05). Conclusion:Hyperbaric oxygen has inhibitory effects on lymphocytes in vitro ,and NAC has preventive effect to some extent.