目的 研究人參皂苷Rg₁對移植性白血病模型小鼠的作用。方法 采用免疫磁性分選法（MACS）從K562細胞中分離、純化CD34⁺CD38^-人白血病干細胞（CD34⁺CD38^- LSCs），流式細胞術檢測分選細胞純度，臺盼藍染色測定分選細胞活性。將6～8周雌性NOD/SCID小鼠27只隨機分為對照組、模型組、人參皂苷Rg₁（200 mg/kg）組，每組9只，模型組和人參皂苷Rg₁組通過尾iv移植CD34⁺CD38^- LSCs構建白血病小鼠模型，ip給藥30 d。觀察各組小鼠一般情況及腹部包塊變化；全自動血常規(guī)檢測儀檢測外周血白細胞、紅細胞、血紅蛋白、血小板水平；免疫組化法檢測肝、脾臟病理學變化；流式細胞儀分析骨髓細胞周期；CCK-8法檢測各組骨髓細胞的增殖能力；細胞免疫熒光染色鑒定骨髓細胞CD34陽性表達情況。結果 分選前K562細胞中CD34⁺CD38^- LSCs細胞群百分比為（9.64±1.14）%，分選后CD34⁺CD38^- LSCs純度可達（96.45±1.63）%；臺盼藍染色顯示分選后細胞活性為（95.26±2.16）%。與對照組比較，模型組小鼠腹部包塊明顯，生存情況較差，體質量顯著下降（P<0.05）；白細胞數(shù)顯著增高，紅細胞數(shù)、血紅蛋白水平、血小板數(shù)均顯著下降（P<0.05）；肝、脾臟結構被破壞，有大量白細胞浸潤；骨髓細胞周期檢測顯示G₀/G₁期比例顯著下降（P<0.05），S期比例顯著升高（P<0.05）；骨髓細胞增殖活力顯著升高（P<0.05）。與模型組比較，人參皂苷Rg₁組小鼠腹部包塊明顯減小，生存情況好轉，體質量顯著增加（P<0.05）；白細胞總數(shù)顯著下降（P<0.05），紅細胞數(shù)、血紅蛋白水平、血小板數(shù)均顯著升高（P<0.05）；肝、脾臟結構明顯恢復；骨髓細胞G₀/G₁期比例顯著升高（P<0.05），G₂/M期和S期比例顯著下降（P<0.05）。細胞免疫熒光檢測顯示，模型組和人參皂苷Rg₁組中小鼠的骨髓細胞中存在大量的人源白血病細胞。結論 NOD/SCID小鼠尾iv移植CD34⁺CD38^- LSCs可成功構建急性髓系白血病小鼠模型，人參皂苷Rg₁能有效緩解NOD/SCID小鼠的白血病癥狀。

Objective To study the effect of ginsenoside Rg₁ on transplanted leukemia mice model. Methods CD34⁺CD38^- LSCs were isolated and purified from K562 cells by immunomagnetic separation (MACS). The purity of CD34⁺CD38^- LSCs was determined by flow cytometry, and the activity of CD34⁺CD38^- LSCs was determined by Trypan blue staining. Totally 36 female NOD/SCID mice, 6—8 weeks old, were randomly divided into control group, model group and ginsenoside Rg₁ (200 mg/kg) group, with nine mice in each group. The mouse model of leukemia in model group and ginsenoside Rg₁ group was established by tail IV transplantation of CD34⁺CD38^- LSCs, and ip administration of corresponding drugs for 30 days. The general condition and abdominal mass changes of mice in each group were observed. Automatic blood routine detector was used to detect peripheral blood white blood cell, red blood cell, hemoglobin, platelet levels. The pathological changes of liver and spleen were detected by immunohistochemistry. Bone marrow cell cycle was analyzed by flow cytometry. CCK-8 method was used to detect the proliferation of bone marrow cells in each group. CD34 positive expression of bone marrow cells was determined by immunofluorescence staining. Results The percentage of CD34⁺CD38^- LSCs in K562 cells was (9.64±1.14)% before separation, and the purity of CD34⁺CD38^- LSCs reached (96.45±1.63)% after separation. Trypan blue staining showed that the cell activity after separation was (95.26±2.16)%. Compared with control group, abdominal mass was obvious in model group, the survival was poor, and body weight was significantly decreased (P<0.05). WBC count was significantly increased, RBC count, hemoglobin level and platelet count were significantly decreased (P<0.05). The structure of liver and spleen was destroyed, and there were a lot of leukocytes. Bone marrow cell cycle detection showed that the proportion of G₀/G₁ phase was significantly decreased (P<0.05), and that of S phase was significantly increased (P<0.05). The proliferation activity of bone marrow cells was significantly increased (P<0.05). Compared with model group, abdominal mass in ginsenoside Rg₁ group was significantly reduced, survival was improved, and body weight was significantly increased (P<0.05). The total number of WBC was significantly decreased (P<0.05), while the RBC number, hemoglobin level and platelet count were significantly increased (P<0.05). The structure of liver and spleen recovered significantly. The proportion of G₀/G₁ phase was significantly increased (P<0.05), while the proportion of G₂/M and S phase was significantly decreased (P<0.05). Immunofluorescence assay showed that there were a large number of human leukemia cells in bone marrow cells of mice in model group and ginsenoside Rg₁ group. Conclusion Tail iv transplantation of CD34⁺CD38^- LSCs construct acute myeloid leukemia mouse model, ginsenoside Rg₁ can effectively alleviate the symptoms of leukemia in NOD/SCID mice.

R285.5

國家自然科學基金資助項目（81860038）；云南省教育廳科學研究基金資助項目（2020J1055）