蝎素组分Ⅲ对Jurkat细胞人白细胞分化抗原25表达的影响

Effects of scorpion venom component Ⅲ on the express of CD25 in Jurkat cells

目的研究蝎素组分Ⅲ(SVC-Ⅲ)对Jurkat细胞人白细胞分化抗原25(CD25)表达的影响。方法用0.1μg.L-1、1μg.L-1和10μg.L-1的SVC-Ⅲ分别作用CD4+T淋巴细胞系Jurkat E6-1细胞,利用逆转录多聚酶链反应(RT-PCR)法检测T细胞活化连接蛋白(LAT)和zeta链相关蛋白-70(ZAP-70)mRNA的表达;利用流式细胞术检测T淋巴细胞活化早期表面标志细胞分化抗原CD25分子表达变化情况。结果与对照组比较,0.1μg.L-1组和1μg.L-1组LAT/GAPDH比值增高(P<0.05),10μg.L-1组LAT/GAPDH比值增高,但差异无统计学意义(P>0.05),3组ZAP-70/GAPDH比值及CD25阳性细胞率均增高(P<0.05);与0.1μg.L-1组比较,1μg.L-1组LAT/GAPDH、ZAP-70/GAPDH比值和CD25阳性细胞率均增高(P<0.05),10μg.L-1组LAT/GAPDH,ZAP-70/GAPDH比值和CD25阳性细胞率均降低(P<0.05),且10μg.L-1组LAT/GAPDH、ZAP-70/GAPDH比值和CD25阳性细胞率较1μg.L-1组也降低(P<0.05)。结论SVC-Ⅲ对Jurkat细胞CD25的表达具有明显的促进作用,其中1μg.L-1SVC-Ⅲ浓度促进作用最强,为蝎素在临床上的合理应用提供一定的实验基础。

Objective To investigate the effects of scorpion venom component Ⅲ （ SVC- Ⅲ） on the expression of CD25 in Jurkat cells. Methods After CD^4＋ T lymphocyte series -Jurkat E6-1 cells were stimulated by Svc-m （0. 1 μg.L^-1, 1μg.L^-1 and 10 μg . L^-1 ）, the expressions of linker for activation of T cell（LAT） and zeta-associated protein-70（ ZAP-70 ） mRNA were checked by reverse transcription polymcrase chain reaction （RT-PCR）- The expression of CD25 （ cluster of differentiation） as a marker of early activation on the surface of T lymphocytes was detected by flow cytometry. Results The value of LAT/GAPDH in 0.1 μg .L^-1 group and μg.L^-1 group was significantly higher than that in control group（P 〈 0.05 ）;The value of LAT/GAPDH in 10μg .L^-1 was higher than control group, but there was no statistical difference （P 〉 0. 05）. The values of ZAP-70/GAPDH and positive rate of CD25 cells in three groups （0. 1μg.L^-1 group, μg.L^-1, 10 μg.L^-1 group） were significantly higher than thoes in control group （P 〈 0.05 ）. The values of LAT/GAPDH, ZAP-70/ GAPDH and positive rate of CD25 cells in 0. 1 μg.L^-1 group were significantly higher but in 10 μg. L^-1 group were significantly lower than thoes in 0. 1μg.L^-1 group （P 〈 0.05 ） ;.The values of LAT/GAPDH, ZAP-70/GAPDH and positive rate of CD25 cells in 10 μg.L^-1 group were significantly lower than thoes in 1 μg . L^-1 group （ P 〈 0.05 ）. Conclusion SVC- Ⅲ can enhance the expression of CD25 in Jurkat cells and 1 μg . L^-1 SVC-Ⅲ was the strongest. This result could provide some experimental bases of reasonable use of scorpion venom in clinical practical.