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研究辛二酰苯胺异羟肟酸(SAHA)对小鼠淋巴细胞活化、增殖和凋亡的影响,并对其免疫调控机制进行初步探讨。以MTS检测淋巴细胞的增殖情况;以双色荧光抗体染色技术结合流式细胞术检测SAHA对T淋巴细胞早期活化抗原CD69表达的影响;采用碘化丙锭(PI)染色检测SAHA对淋巴细胞周期的影响;以Annexin V-PE染色分析细胞凋亡;应用免疫印迹检测SAHA对Cleaved caspase-3表达的影响。结果显示,SAHA对Con A刺激的淋巴细胞增殖具有抑制作用,其IC50为0.30±0.06μmol/L;SAHA对淋巴细胞的早期活化抗原CD69的表达具有明显的抑制作用(P<0.05);PI染色、Annexin V-PE染色和免疫印迹结果均显示,SAHA对小鼠淋巴细胞的凋亡具有促进作用,且呈剂量依赖性。研究表明,SAHA可能通过抑制小鼠淋巴细胞的活化、增殖以及促进其凋亡而发挥免疫调控功能。
To study the effect of suberoylanilide hydroxamic acid (SAHA) on the activation, proliferation and apoptosis of mouse lymphocytes, and to investigate its immunomodulatory mechanism. The proliferation of lymphocytes was detected by MTS. The effect of SAHA on CD69 expression of early activation antigen of T lymphocytes was detected by dual-color fluorescent antibody staining and flow cytometry. The expression of SAHA on lymphocyte cycle was detected by propidium iodide (PI) The apoptosis was analyzed by Annexin V-PE staining. The effect of SAHA on the expression of Cleaved caspase-3 was detected by Western blotting. The results showed that SAHA could inhibit the proliferation of Con A-stimulated lymphocytes with an IC50 of 0.30 ± 0.06 μmol / L. SAHA significantly inhibited the expression of CD69, an early activation antigen of lymphocytes (P <0.05) , Annexin V-PE staining and Western blotting results showed that SAHA can promote lymphocyte apoptosis in a dose-dependent manner. Studies have shown that SAHA may play an immunomodulatory role by inhibiting mouse lymphocyte activation, proliferation and promoting apoptosis.