慢性乙型肝炎患者PBMCFas表达和凋亡的检测及意义

Fas expression and apoptosis of peripheral blood lymphocytes in patients with chronic hepatitis B and its significance

目的 探讨活化诱导细胞死亡 (Activation inducedcelldealth ,AICD)在乙型肝炎发病机制中的意义。方法 分离 30例乙型肝炎患者和 1 2例健康献血员外周血单个核细胞 (PBMC) ,利用间接免疫荧光染色法 ,通过流式细胞仪检测PBMCFas的表达 ;分离2 0例乙型肝炎病人和 1 0例健康献血员PBMC ,在PHA P的刺激下短期培养 ,采用PI染色法 ,经流式细胞仪检测PBMC的凋亡。结果慢性乙型肝炎组PBMCFas表达率 [( 53.31± 1 4 .53) % ]高于正常对照组PBMCFas表达率 [( 39.32± 1 1 .2 8) % ,P <0 .0 5]和慢性重型乙型肝炎组PBMCFas表达率 [( 37.93± 1 5.89) % ,P <0 .0 1 ]。慢性乙型肝炎组PBMC凋亡率 [( 30 .57± 1 3.4 3) % ]高于正常对照组PBMC凋亡率 [( 1 1 .4 5± 5.2 7) % ,P <0 .0 1 ]和慢性重型乙型肝炎组PBMC凋亡率 [( 1 3.59± 6 .4 4) % ,P <0 .0 1 ];HBeAg( + )组PBMC凋亡率高于正常对照组PBMC凋亡率 [( 2 9.50± 1 2 .2 5) %vs( 1 1 .4 5± 5.2 7) % ,P <0 .0 1 ]。结论 HBV感染能诱导慢性乙型肝炎外周血淋巴细胞Fas表达增加 ,通过Fas/FasL途径使Fas+淋巴细胞发生AICD ,这可能是形成HBV慢性感染免疫耐受的一个重要机制。

Objective To study the significance of AICD in the pathogenesis of hepatitis B. Methods The peripheral blood mononuclear cells (PBMC) were isolated from 30 patients with hepatitis B and 12 healthy blood donors. The expression of Fas was assayed with flow cytometry. The peripheral blood mononuclear cells were isolated from 20 patients with hepatitis B and 10 healthy blood donors. Then the PBMC were cultured with PHA P for 72 hours in vitro . The apoptosis of the PBMC was stained with PI and assayed with flow cytometry. Results The expression rate of Fas of PBMC in chronic hepatitis B [(53.31±14.53)%] was significantly higher than that in healthy blood donors [(39.32±11.28)%, P < 0.05] and chronic severe hepatitis B [(37.93±15.89)%, P <0.01]. The apoptosis rate of PBMC in chronic hepatitis B [(30.57± 13.43)%] was significantly higher than that in healthy blood donors [(11.45±5.27)%, P <0.01] and in chronic severe hepatitis B [(13.59±6.44)%, P <0.01] . The apoptosis rate of PBMC in HBeAg(+) patients was significantly higher than that in healthy blood donors [(29.50± 12.25)% vs (11.45±5.27)%, P <0.01]. Conclusion These results show that the increase of Fas expression of PBMC can be induced in hepatitis B infection. AICD of Fas positive lymphocytes may be induced by Fas/FasL. AICD may be an important mechanism in the formation of immune tolerance in HBV chronic infection.