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目的:研究益气养阴方及其拆方后的扶正方、祛邪方对人急性髓系白血病细胞凋亡及细胞色素C(Cytochrome C,Cyt-C),凋亡蛋白酶活化因子(apoptotic protease activating factor 1,Apaf-1),第二个线粒体衍生的半胱天冬蛋白酶激活因子(the second mitochondria-derived activator of caspase/direct IAP-blinding protein with low PI,Smac/Diablo),凋亡诱导因子(apoptosis-inducing factors,AIF)表达的影响,探讨益气养阴方治疗白血病的可能作用机制。方法:采用NOD/SCID小鼠,以KG-1a细胞株建立人急性髓系白血病模型,随机分为全方组、祛邪组、扶正组和空白组,取NOD/SCID小鼠脾细胞制成细胞悬液,应用流式细胞术检测NOD/SCID小鼠细胞凋亡率;采用免疫组化法检测NOD/SCID小鼠骨髓中线粒体相关凋亡因子CytC,Apaf-1,Smac/Diablo和AIF的表达。结果:用药后,用药组小鼠生存时间较空白组显著延长(P<0.01),全方组小鼠生存时间较扶正组与祛邪组显著延长(P<0.01),祛邪组小鼠生存时间较扶正组显著延长(P<0.01)。用药组细胞凋亡率较空白组明显升高(P<0.01),全方组较扶正组与祛邪组显著升高(P<0.01),祛邪组较扶正组显著升高(P<0.01)。用药组小鼠骨髓中Cyt-C,Apaf-1,Smac/Diablo,AIF的表达较空白组显著升高(P<0.05),其中Cyt-C与Apaf-1表达显著升高(P<0.01);全方组均高于扶正组与祛邪组(P<0.01);祛邪组较扶正组均升高(P<0.05),Cyt-C,Apaf-1和Smac/Diablo表达显著升高(P<0.01)。其中对于各指标的影响全方组均优于扶正组与祛邪组,祛邪组均优于扶正组。结论:益气养阴方能够提高NOD/SCID小鼠的细胞凋亡率,上调线粒体相关凋亡因子Cyt-C,Apaf-1,Smac/Diablo,AIF的表达,并诱导其凋亡,抑制白血病细胞增殖,其机制可能与线粒体凋亡通路有关。
Objective: To study the effects of Qi-Qiyin-Yin Recipe and Fuzheng Prescription, Fangzheng Prescription, and Quxie Recipe on the apoptosis of human acute myeloid leukemia and the effects of Cytochrome C (Ct), apoptotic protease activating factor 1, Apaf-1), the second mitochondria-derived activator of caspase / direct IAP-blinding protein with low PI (Smac / Diablo), apoptosis inducing factor (apoptosis-inducing factors, AIF) expression, to explore the possible mechanism of Yiqi Yangyin Prescription for the treatment of leukemia. Methods: Acute myeloid leukemia model was established by KG-1a cell line in NOD / SCID mice and randomly divided into the whole group, Quxie group, Fuzheng group and blank group. The NOD / SCID mice spleen cells The apoptosis rate of NOD / SCID mice was detected by flow cytometry. The mitochondria-associated apoptotic factors CytC, Apaf-1, Smac / Diablo and AIF in NOD / SCID mice bone marrow were detected by immunohistochemistry expression. Results: After treatment, the survival time of mice in treatment group was significantly longer than that in control group (P <0.01). The survival time of mice in treatment group was significantly longer than that in Fuzheng group and Quxie group (P <0.01) Time than Fuzheng group was significantly longer (P <0.01). The apoptosis rate of the treated group was significantly higher than that of the blank group (P <0.01), that of Fufang group and Quxie group (P <0.01), Quxu group was significantly higher than that of Fufang group (P <0.01) ). The expression of Cyt-C, Apaf-1, Smac / Diablo and AIF in the bone marrow of the mice in treatment group was significantly higher than that in the blank group (P <0.05), and the expressions of Cyt-C and Apaf- (P <0.01); Quxie group was higher than that of Fufang group (P <0.05); the expression of Cyt-C, Apaf-1 and Smac / Diablo was significantly increased P <0.01). Among them, the influence of each index is better than that of Fuzheng and Quxie groups, Quxie group are superior to Fuzheng group. Conclusion: Yiqi Yangyin Prescription can increase the apoptosis rate of NOD / SCID mice and up-regulate the expression of Cyt-C, Apaf-1, Smac / Diablo and AIF in mitochondria, induce apoptosis and inhibit leukemia Cell proliferation, the mechanism may be related to mitochondrial apoptosis pathway.