The Study On The Mechanism Of Inducing KG-1a Cell Apoptosis By Yiqi Yangyin Recipe And Its Different Assembling Through Mitochondrial Pathway

Objective: Taking the NOD/SCID mice as the research platform and KG-1a cells were used to establish the model of human acute myeloid leukemia to explore the possible mechanism of inducing the apoptosis of the acute myeloid leukemia KG-1a cells by Yiqi Yangyin Recipe(supplementing qi and nourishing yin) and its different assembling through mitochondrial pathway. In order to seek for the experimental support that Chinese medicine induces the myeloid leukemia KG-1a cells apoptosis through mitochondrial pathway, in a hope of providing a theoretical basis for studying the new anti-leukemia medicine under the guidance of this theory.Methods: Taking the NOD/SCID mice as the experimental subjects and injected with KG-1a cells from the tail vein after irradiation by 60 Co to establish the model of acute myeloid leukemia. The model mice were divided into control group, Fuzheng group, Quxie group, Quanfang group, while the non-modeling mices were as the blank control group. Observing the general living conditions of mices, moribund mices was tested by bone marrow morphology and histopathology. After treating four weeks, killing all the mices and taking the spleen cell suspensions of the mice to measure the cell cycle and cell apoptosis rates by the flow cytometry. The immunohistochemistry was used to detect the expression of Cyt-C、Apaf-1、Smac/Diablo and AIF.Results: After drug treatment, the survival time of treatment group and the blank control group were significantly longer than the control group(P <0.01), the survival time of Quanfang group and blank control group of mice compared with Fuzheng group and Quxie group were longer(P <0.01), Quanfang group mice compared with the blank control group were not statistically significant(P>0.05), survival time of Quxie group was longer than Fuzheng group mice(P <0.01). Cells in the G1 phase and apoptosis rate in all treatment groups were significantly higher than the control group and blank control group(P <0.01), Quanfang group is higer than Fuzheng group and Quxie group(P <0.01), Quxie group cells in the G1 phase was slightly higher than Fuzheng group(P <0.05), Quxie group cells in the cell apoptosis rate was higer than Fuzheng group(P <0.01). In the control group and blank control group showed no statistically significant(P>0.05). The expression of Cyt-C, Apaf-1, Smac / Diablo and AIF in mouse bone marrow,the treatment group were higher than that in the control group(P<0.05), which was significantly higher Cyt-C and Apaf-1 expression(P <0.01);All Quanfang group were higher than Fuzheng group and Quxie group group(P <0.01); Quxie group was increased than Fuzheng group(P<0.05), the expression of Cyt-C, Apaf-1 and Smac / Diablo was significantly higher(P <0.01). For each indicators wastested, Quanfang group was all better than Fuzheng group and Quxie group, Quxie group were better than Fuzheng group.Conclusion: The results showed that Yiqi Yangyin Recipe can regulate cell cycles, inhibitit the proliferation of leukemia cells, and improve the expression on the mitochondrial-related factors Cyt-C, Apaf-1, Smac/Diablo and AIF, to induce apoptosis of and increase the rate of apoptosis of the leukemia cells, Thereby prolonging the survival time of leukemic mice and revealing that Yiqi Yangyin Recipe has signifinant therapeutic effect on acute myeloid leukemia.