升麻鳖甲汤调控线粒体融合相关蛋白Mfn2、Opa1促进急性白血病细胞凋亡作用机制  

Mechanism of Shengma Biejia Decoction in regulating mitochondrial fusion related proteins Mfn2 and Opal to promote apoptosis of acute leukemia cells

作　　者：魏文健 崔思远[2,3,4] 司雨平 马国栋 殷学伟 李宗宏 王琰 郑伟[2] 徐杰[2] 刘月莉[2] 王敬毅 徐瑞荣[2] WEI Wenjian;CUI Siyuaann;SI Yuping;MA Guodong;YIN Xuewei;LI Zonghong;WANG Yan;ZHENG Wei;XU Jie;LIU Yueli;WANG Jingyi;XU Ruirong(The First Clinical School of Shandong University of Traditional Chinese Medicine,Jinan 250355,China;Affliated Hospital of Shandong University of Traditional Chinese Medicine,Jinan 250014,China;Shandong Laibo Biological Technology Co.,LTD,Jinan 250101,China;Clinical Medical College of Shandong University,Jinan 250013,China)

机构地区：[1]山东中医药大学第一临床医学院,济南250355 [2]山东中医药大学附属医院,济南250014 [3]山东莱博生物科技有限公司,济南商250101 [4]山东大学临床医学院,济南250013

出　　处：《中华中医药杂志》2024年第8期3943-3948,共6页China Journal of Traditional Chinese Medicine and Pharmacy

基　　金：国家自然科学基金项目(No.81903984);山东省自然科学基金项目(No.ZR2019PH018);山东省医药卫生科技发展计划项目(No.202003040045,No.2018WS184)。

摘　　要：目的:探讨升麻鳖甲汤干预急性白血病的作用机制,尤其是在调控线粒体融合相关蛋白及促进细胞凋亡方面的作用。方法:48只NSG小鼠随机分为4组:空白对照组、模型组、全方组、拆方组,每组12只,除空白对照组外其余3组小鼠均经尾静脉注射KG-1a细胞建模。空白对照组与模型组小鼠予0.9%氯化钠溶液灌胃,全方组和拆方组小鼠分别予升麻鳖甲汤及其拆方药液灌胃,均灌胃28 d。持续监测各组小鼠生存状态、生存时间。HE染色法观察各组小鼠肝脏、脾脏、骨髓组织病理变化;采用免疫组织化学染色法检测细胞凋亡相关蛋白Bcl-2相关X蛋白(Bax)、骨髓细胞白血病1蛋白(Mcl-1)表达水平;免疫荧光技术、蛋白免疫印迹法检测线粒体融合相关蛋白线粒体融合蛋白2(Mfn2)、视神经萎缩蛋白1(Opa1)表达水平。结果:升麻鳖甲汤及其拆方较模型组能明显改善小鼠生存状态,延长小鼠生存时间(P<0.05),细胞凋亡相关蛋白Bax表达水平升高,Mcl-1表达水平下降,线粒体融合相关蛋白Mfn2、Opa1表达水平下降(P<0.05),且全方组较拆方组趋势更为显著。结论:升麻鳖甲汤干预急性白血病的作用机制可能与下调白血病细胞线粒体融合相关蛋白Mfn2和Opa1的表达,从而促进细胞凋亡有关。Objective:To explore the mechanism of Shengma Biejia Decoction in treating acute leukemia(AL),especially in regulating mitochondrial fusion related proteins and promoting apoptosis.Methods:Forty-eight NSG mice were randomly divided into 4 groups:blank control group,model group,whole recipe group,disassembled recipe group,with 12 mice in each group.Except the blank control group,the other three groups of mice were injected with KG-la cells through tail vein for modeling.Mice in model group and blank control group were given normal saline by gavage,the whole recipe group and the disassembled recipe group were given Shengma Biejia Decoction and its disassembled recipe solution by gavage respectively.all for 28 days.The survival status and survival time of mice in each group were continuously monitored.HE staining was used to observe the pathological changes of liver,spleen and bone marrow of mice in each group;Immunohistochemical staining was used to detect the expression level of apoptosis related proteins Bax and Mcl-1;Immunofluorescence and Western Blot were used to detect the expression level of mitochondrial fusion related proteins Mfn2 and Opal.Results:Compared with the model group,after the intervention of Shengma Biejia Decoction and its disassembled formula,the survival status of mice was significantly improved,the survival time of mice was prolonged(P<0.05),the expression level of apoptosis related protein Bax increased,the expression level of Mcl-1 protein decreased and the expression level of mitochondrial fusion related proteins Mfn2 and Opal decreased(P<0.05),the trend of the whole formula group of Shengma Biejia Decoction was more significant than that of the disassembled formula group.Conclusion:Shengma Biejia Decoction has the effect of intervention in AL,and its mechanism may be related to the down-regulation of the expression of mitochondrial fusion related proteins Mfn2 and Opal in leukemia cells,thus promoting cell apoptosis.

关 键 词：升麻鳖甲汤 线粒体动力学 线粒体融合 细胞凋亡 急性白血病 线粒体融合蛋白2 视神经萎缩蛋白1 

分 类 号：R285[医药卫生—中药学]