加味青蒿鳖甲汤对急性髓系白血病缓解后CD34^(+)细胞源DC共刺激分子B7-1、B7-2的影响  

Effect of Jiawei Qinghao Biejia Decoction on CD34+cell-derived DC costimulatory molecules B-and B-in AML-CR patients

作　　者：朱国庆 彭名行 雷斯 尹尚瑾 陈孟豪 左乐 黄礼明 ZHU Guo-qing;PENG Ming-hang;LEI Si;YIN Shang-Jin;CHEN Meng-hao;ZUO Le;HUANG Li-ming(Guizhou University of Traditional Chinese Medicine,Guiyang Guizhou,550025,China;The Second Affiliated Hospital of Guizhou University of Traditional Chinese Medicine,Guiyang Guizhou,550003,China)

机构地区：[1]贵州中医药大学,贵州贵阳550025 [2]贵州中医药大学第二附属医院,贵州贵阳550003

出　　处：《时珍国医国药》2024年第12期2725-2732,共8页Lishizhen Medicine and Materia Medica Research

基　　金：国家自然科学基金(81860801);贵州中医药大学研究生教育创新计划项目(YCXZRB202210,YCXZRB202206)。

摘　　要：目的探讨加味青蒿鳖甲汤治疗急性髓系白血病完全缓解(AML-CR)的可能作用机制。方法将12只新西兰大白兔用随机数字表法分为正常组和加味青蒿鳖甲汤高、中、低剂量组,每组3只。加味青蒿鳖甲汤高、中、低剂量组兔分别给予加味青蒿鳖甲汤浓缩煎剂30、15、7.5g/(kg·d)灌胃3d,正常组兔灌胃等体积蒸馏水,末次给药后心尖取血,获得正常兔血清和加味青蒿鳖甲汤高、中、低剂量兔血清。将AML-CR患者骨髓稀释后采用免疫磁珠法分离提纯CD34^(+)细胞,并对CD34^(+)细胞进行扩增,将扩增后的CD34^(+)细胞分为加味青蒿鳖甲汤高、中、低剂量血清组、正常血清组、细胞因子组以及空白组。除空白组外,其余各组均添加细胞因子(GM-CSF+IL-4+IFN-α+TNF-α)诱导9d成为树突细胞(DC),培养过程中用倒置显微镜观察细胞形态。用FCM检测DC表面抗原分子B7-1、B7-2、CD83、CD1a、HLA-DR表达率,用qRT-PCR、Western blot检测DC中B7-1、B7-2蛋白及mRNA表达。结果空白组DC细胞始终未观察到树突状改变;加味青蒿鳖甲汤高、中、低剂量组、细胞因子组和正常血清组细胞胞体显著增大,细胞表面出现星状多形性和树枝状突起。与空白组相比较,加味青蒿鳖甲汤高、中、低剂量组DC表面标志物B7-1、B7-2、CD83、CD1a、HLA-DR表达均显著升高(P<0.05);与细胞因子组和正常血清组相比较,加味青蒿鳖甲汤高、中、低剂量组DC表面标志物B7-1、B7-2、CD1a、HLA-DR表达均明显升高(P<0.05);加味青蒿鳖甲汤高、中剂量组DC表面标志物CD83明显升高(P<0.05)。空白组、细胞因子组和正常血清组三组间B7-1和B7-2 mRNA及蛋白相对表达无明显差异(P>0.05);与空白组、细胞因子组和正常血清组比较,加味青蒿鳖甲汤高、中、低剂量血清组B7-1和B7-2 mRNA及蛋白相对表达均显著升高(P<0.05)。结论加味青蒿鳖甲汤治疗AML-CR的作用机制可能与该方能够有效促进AML-CRObjective To explore the possible mechanism of Jiawei Qinghao Biejia Decoction in treating Complete remission,(CR)of acute myeloid leukemia,(AML).Methods Twelve New Zealand white rabbits were randomly divided into normal group and high,middle and low dose groups of Jiawei Qinghao Biejia Decoction,with 3 rabbits in each group.The rabbits in the high,middle and low dosage groups of Jiawei Qinghao Biejia Decoction were given concentrated decoction of Jiawei Qinghao Biejia Decoction 30g,15g and 7.5g/(kg.d)by gavage for 3 days,while the rabbits in the normal group were given the same volume of distilled water by gavage,and blood was taken from the apex after the last administration to obtain normal rabbit serum and high,middle and low dosage rabbit serum of Jiawei Qinghao Biejia Decoction.The CD34^(+)cells of AML-CR patients were isolated and purified by immunomagnetic beads method,and the CD34^(+)cells were amplified.The amplified CD34^(+)cells were divided into high,medium and low dose serum groups,normal serum groups,cytokine groups and blank groups.Except the blank group,all the other groups were induced to become dendritic cells,(DC)by adding cytokines(GM-CSF+IL-4+IFN-α+TNF-α)for 9 days,and the cell morphology was observed by inverted microscope during the culture.The expression rates of DC surface antigen molecules CD80,CD86,CD83,CD1a and HLA-DR were detected by FCM,and the expressions of B7-1,B7-2 protein and mRNA in DC were detected by qRT-PCR and Western blot.Results Dendritic changes were never observed in DC cells in blank group.The cell bodies in the high,middle and low dose groups,cytokine group and normal serum group of Jiawei Qinghao Biejia Decoction were significantly enlarged,and stellate pleomorphism and dendritic processes appeared on the cell surface.Compared with the blank group,the expressions of DC surface markers B7-1,B7-2,CD83,CD1a and HLA-DR in the high,middle and low dose groups of Jiawei Qinghao Biejia Decoction increased significantly(P<0.05).Compared with the cytokine group and the norm

关 键 词：急性髓系白血病 加味青蒿鳖甲汤 树突细胞 共刺激分子 抗原提呈 

分 类 号：R285.5[医药卫生—中药学]