“痰瘀同治”法对再生障碍性贫血小鼠骨髓有核细胞凋亡的影响及其机制

"Comanagement of Phlegm and Stasis" on the Apoptosis of Marrow Karyocyte in Mice with Aplastic Anemia and its Possible Mechanism

目的:探讨"痰瘀同治"法对再生障碍性贫血小鼠骨髓有核细胞凋亡的影响及其相关机制。方法:复制免疫介导的再生障碍性贫血(AA)小鼠模型,取其血清诱导正常小鼠骨髓有核细胞的凋亡过程;同时用中药(单纯补肾、补肾祛痰瘀)灌给的大鼠血清作为治疗因素,体外共培养24h,观察有核细胞凋亡率、细胞内Caspase-3酶活性及Bax蛋白表达水平。结果:AA小鼠血清处理后,正常小鼠骨髓有核细胞凋亡率显著提高(P<0.01),两治疗组均能显著减轻AA小鼠血清的凋亡诱导作用(P<0.01),抑制AA血清诱导下骨髓有核细胞内Caspase-3酶活性(P<0.05),且以补肾祛痰方药的作用更明显(P<0.05);两治疗组均能显著降低骨髓细胞的Bax蛋白阳性强度与阳性率乘积(P<0.05),但两组间无显著性差异。结论:AA小鼠血清能显著诱导骨髓造血细胞的凋亡过程,两治疗组均能通过Caspase-3/Bax途径起到抑制作用,且以补肾祛痰瘀较为显著,但两者疗效的差异与Bax无显著相关性。

Objective： To explore the ＂Comanagement of Phlegm and Stasis＂ on the apoptosis of marrow karyocyte treated with aplastic anemia serum and its possible mechanism.Methods： Immune-mediated model of aplastic anemia（AA） mice was copied,and their serum was used to induce the apoptosis process of marrow karyocyte from normal mice.After co-culture with serum from rat administrated with Traditional Chinese Medicine（Prescription on reinforcing kidney purely or both on reinforcing kidney and dispel phlegm and blood stasis） for 24 hours,the apoptosis rate,enzymic activity of caspase-3 and the expression of Bax protein of karyocyte were detected.Results： The apoptosis rate of marrow karyocyte is significantly higher in AA mice serum treated group than that not（P〈0.01）,both the treated group can relieve the apoptosis inducing effect of AA mice serum（P〈0.01）,inhibit the enzymic activity of Caspase-3（P〈0.05）,and in which,the group treated both on ＂reinforcing kidney and dispel phlegm and blood stasis＂ is more obviously（P〈0.05）.The multiply of positive staining intensity and rate of Bax protein is down-regulated in both treated groups（P〈0.05）,while no significance was observed.Conclusion： The apoptosis process can be induced by serum from AA mice,and can be inhibited by both two prescriptions through the caspase-3/Bax pathway.This phenomenon is more noted in group treated ＂both on reinforcing kidney and dispel phlegm and blood stasis＂,and had no relationship to Bax protein expression.