回阳固本汤促进异基因造血干细胞移植后自然杀伤细胞重建的机制研究  

Mechanism of Huiyang Guben Decoction in Promoting Natural Killer Cell Reconstruction After Allogeneic Hematopoietic Stem Cell Transplantation

作　　者：高小丽 何海涛 王启 姚翔媚 秦涛 张海溪[1] 赵婕 冯帅[1] 胡芃[1] 杨同华[1] GAO Xiaoli;HE Haitao;WANG Qi;YAO Xiangmei;QIN Tao;ZHANG Haixi;ZHAO Jie;FENG Shuai;HU Peng;YANG Tonghua(Dept.of Hematology,the First People's Hospital of Yunnan,Yunnan Clinical Research Center for Hematologic Disease,Kunming 650032,China)

机构地区：[1]云南省第一人民医院血液科云南省血液系统疾病临床医学研究中心,昆明650032

出　　处：《中国医院用药评价与分析》2023年第9期1046-1051,共6页Evaluation and Analysis of Drug-use in Hospitals of China

基　　金：国家自然科学基金(No.82060810);云南省血液系统疾病临床医学研究中心开放项目(No.2020LCZXKF-XY06,No.2020LCZXKF-XY12,No.2020LCZXKF-XY16,No.2020LCZXKF-XY18,No.2021LCZXXF-XY02,No.2021LCZXXF-XY03);云南省科技厅-昆明医科大学应用基础研究联合专项基金资助项目(No.202201AY070001-244)。

摘　　要：目的:探讨回阳固本汤对异基因造血干细胞移植(allo-HSCT)后自然杀伤细胞(NK细胞)重建与NK细胞活性的影响及其分子调控机制。方法:(1)临床研究。选取2021年7月至2022年9月于该院行allo-HSCT的患者40例,根据治疗方式分为对照组(n=20)与HYGB组(n=20)。对照组患者仅采用allo-HSCT单独治疗,HYGB组患者于allo-HSCT前采用回阳固本汤干预4周。allo-HSCT治疗后对两组患者进行疗效评估,检测两组患者外周血NK细胞计数、外周血NK细胞标志分子CD56与CD16的表达,分离两组患者外周血NK细胞,体外培养测定细胞活性,检测磷脂酰肌醇3激酶/蛋白激酶B(PI3K/Akt)通路蛋白表达情况。(2)体外细胞实验。从健康体检者外周血中分离培养NK细胞,分为对照组(阴性对照)、HYGB组(回阳固本汤干预)与LY294002组(回阳固本汤+PI3K抑制剂LY294002干预),测定三组细胞活性与凋亡情况。结果:(1)对照组患者造血重建良好率为90%(18/20),复发率为10%(2/20),非移植相关死亡率为5%(1/20);HYGB组患者造血重建良好率为100%(20/20),复发率为5%(1/20),未出现死亡,两组患者疗效比较,差异无统计学意义(P>0.05)。但HYGB组患者急性移植物抗宿主病(aGVHD)发生率(10%,2/20)显著低于对照组(45%,9/20),外周血NK细胞比例及CD56、CD16表达量显著高于对照组,NK细胞体外增殖活性高于对照组,凋亡率低于对照组,差异均有统计学意义(P<0.05)。与对照组相比,HYGB组患者NK细胞内PI3K/Akt通路显著激活,差异有统计学意义(P<0.05)。(2)与HYGB组相比,LY294002组NK细胞增殖活性降低,凋亡率升高,差异均有统计学意义(P<0.05),LY294002逆转了回阳固本汤在体外促进NK细胞活性的效果。结论:回阳固本汤通过激活PI3K/Akt通路促进allo-HSCT后NK细胞重建与活性维持。OBJECTIVE:To probe into the effects of Huiyang Guben decoction on natural killer(NK)cell reconstruction after allogeneic hematopoietic stem cell transplantation(allo-HSCT)and its molecular mechanism.METHODS:(1)Clinical experiment.Totally 40 patients undergoing allo-HSCT in the hospital from Jul.2021 to Sept.2022 were divided into the control group(n=20)and HYGB group(n=20)according to the treatment method.The HYGB group was intervened with Huiyang Guben decoction for 4 weeks before allo-HSCT.The efficacy of two groups was evaluated after allo-HSCT treatment.The peripheral blood NK cells were counted,the expression of peripheral blood NK cell marker molecules CD56 and CD16 was detected.The peripheral blood NK cells of two groups were isolated,the cell viability was measured in vitro,and the expression of phosphatidylinositol 3 kinase/protein kinase B(PI3K/Akt)pathway protein was detected.(2)In vitro cell experiment.NK cells were isolated and cultured from the peripheral blood of healthy people,and then divided into the control group(negative control),HYGB group(intervention with Huiyang Guben decoction)and LY294002 group(intervention with Huiyang Guben decoction+PI3K inhibitor LY294002).The cell viability and apoptosis of three groups were measured.RESULTS:(1)The good rate of hematopoietic reconstruction in the control group was 90%(18/20),the recurrence rate was 10%(2/20),and the nontransplantation-related mortality was 5%(1/20).The good rate of hematopoietic reconstruction in the HYGB group was 100%(20/20),the recurrence rate was 5%(1/20),and there was no death.There was no significant difference in efficacy between two groups(P>0.05).However,the incidence of acute graft-versus-host disease(aGVHD)in the HYGB group was 10%(2/20),significantly lower than 45%(9/20)in the control group,the proportion of NK cells in peripheral blood and the expression of CD56 and CD16 were significantly higher than those in the control group,the proliferation activity of NK cells in vitro was higher than that in the control group,and

关 键 词：回阳固本汤 异基因造血干细胞移植 自然杀伤细胞 重建 活性 

分 类 号：R932[医药卫生—生药学]