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研究棉酚(gossypol)对小鼠淋巴细胞体外增殖和凋亡作用的影响及作用机制。分离小鼠淋巴结细胞,以CFSE染色,加入佛波醇酯(PDB)和离子霉素(Ion)进行刺激,以流式细胞仪分析棉酚对淋巴细胞增殖率的影响;以MTS检测淋巴细胞的增殖情况;以Annexin V-PE染色分析细胞凋亡;用Hoechst33342染色观察细胞核形态。CFSE染色分析显示,棉酚对PDB+Ion诱导的小鼠淋巴细胞增殖,具有明显的抑制作用(P<0.01),且呈剂量依赖性。以MTS分析细胞增殖情况,也获得相似的结果。终浓度为4、8和16μmol/L的棉酚处理8 h后的细胞凋亡率分别为4.39%、6.57%和12.14%,PDB+Ion对照组凋亡率为2.13%。凋亡率随着棉酚作用时间的增加而上升,存在时间和剂量依赖关系。高浓度棉酚作用后大量细胞核呈现染色质固缩、核碎裂和致密浓染等凋亡特征。研究表明,棉酚能有效抑制小鼠淋巴细胞体外增殖和诱导其发生凋亡,并呈现出时间和剂量依赖关系。
To study the effect and mechanism of gossypol on proliferation and apoptosis of mouse lymphocytes in vitro. The mouse lymph node cells were isolated, stained with CFSE, stimulated with phorbol ester (PDB) and ionomycin (Ion), and the effects of gossypol on the proliferation of lymphocytes were analyzed by flow cytometry. Proliferation; Cell apoptosis was analyzed by Annexin V-PE staining; Nuclear morphology was observed by Hoechst33342 staining. CFSE staining showed that gossypol had a significant inhibitory effect on PDB + Ion-induced mouse lymphocyte proliferation (P <0.01) in a dose-dependent manner. MTS analysis of cell proliferation, also obtained similar results. The apoptosis rates of Gossypol at the concentrations of 4, 8 and 16μmol / L for 8 h were 4.39%, 6.57% and 12.14%, respectively. The apoptosis rate of PDB + Ion control group was 2.13%. Apoptosis rate increased with the increase of gossypol time, there is a time-and dose-dependent relationship. After exposure to high concentration of gossypol, many nuclei showed chromatin condensation, nuclear fragmentation and dense staining. Studies have shown that gossypol can effectively inhibit mouse lymphocyte proliferation in vitro and induce its apoptosis, showing a time-and dose-dependent relationship.