南凌杉 张雅月 王佳 丁晓庆 郭明 谌海燕 刘军霞 李玲 陈信义 郎海燕

摘要 目的：驗证健脾益气摄血方缓解ITP乏力症状与改善线粒体功能的相关性。方法：通过被动免疫造模法建立ITP小鼠模型，分为正常组、模型组、强的松组、健脾益气摄血（JPYQSX）中、高剂量组。通过光谱法检测小鼠脾脏组织活性氧含量;通过实时荧光定量PCR（qPCR）检测线粒体DNA相对拷贝数;通过免疫荧光法检测结肠ATP含量。结果：与正常组比较，模型组（P<0.01）、强的松组（P<0.05）、健脾益气摄血各剂量组（P<0.01）小鼠脾脏组织ROS含量增多，脾脏组织线粒体DNA相对拷贝数明显下调（P<0.01），结肠组织ATP含量明显下调（P<0.01）;与模型组比较，强的松组、健脾益气摄血各剂量组小鼠脾脏组织ROS含量减少，脾脏组织线粒体DNA相对拷贝数比较无统计学意义（P>0.05），结肠组织ATP含量均显著升高（P<0.01）。结论：健脾益气摄血方可通过改善线粒体功能来有效缓解ITP乏力症状。

关键词 免疫性血小板减少症;乏力;健脾益气摄血;线粒体;线粒体功能;活性氧含量;线粒体DNA;ATP含量

Study on the Mechanism of Jianpi Yiqi Shexue Prescription on Regulating ITP Related Fatigue Based on Mitochondrial Functional Characteristics

NAN Lingshan1，ZHANG Yayue2，Wang Jia3，DING Xiaoqing1，GUO Ming1，CHEN Haiyan1，LIU Junxia1，LI Ling1，CHEN Xinyi2，LANG Haiyan1

（1 Dongfang Hospital，Beijing University of Chinese Medicine，Beijing 100078，China; 2 Dongzhimen Hospital，Beijing University of Chinese Medicine，Beijing 100700，China; 3 Henan University of Traditional Chinese Medicine，Zhengzhou 450003，China）

Abstract Objective：To verify the correlation of the Jianpi Yiqi Shexue Formula in relieving ITP related fatigue and improving mitochondrial function.Methods：ITP mice model was established by the passive immunization mode method and divided into a normal group，a model group，a prednisone group，a Jianpi Yiqi Shexue Formula of medium and high dose group.The reactive oxygen content in mouse spleen tissue was detected by spectroscopy; the relative copy number of mitochondrial DNA was detected by real-time fluorescent quantitative PCR （qPCR）; the ATP content in colon was detected by immunofluorescence.Results：Compared with the normal group，the content of ROS in spleen tissue of mice increased，the relative copy number of mitochondrial DNA in spleen tissue was significantly down-regulated （P<0.01），and the content of ATP in colon tissue was significantly down-regulated （P<0.01） in the model group （P<0.01），in the prednisone group （P<0.05），the Jianpi Yiqi Shexue Formula of each dose group （P<0.01）; compared with the model group，the content of ROS in spleen tissue of mice in the prednisone group （P<0.05），and the Jianpi Yiqi Shexue Formula of each dose group decreased，and the relative copy number of mitochondrial DNA in spleen tissue showed no statistical difference.Conclusion：Jianpi Yiqi Shexue Formula can effectively relieve ITP related fatigue by improving mitochondrial function.

Keywords Immune thrombocytopenia; Fatigue; Jianpi Yiqi Shexue; Mitochondrial; Mitochondrial function; ROS content; Mitochondrial DNA; ATP content

中图分类号：R285.5;R558+.2文献标识码：Adoi：10.3969/j.issn.1673-7202.2021.03.002

免疫性血小板减少症（Immune Thrombocytopenia，ITP）患者乏力症状是该病治疗中亟待解决的关键问题，且长期持续的乏力会严重影响ITP患者生命质量，甚至导致患者情绪低落而发生焦虑事件。但迄今为止，对于导致ITP患者乏力原因尚无明确认识和有效治疗措施。

线粒体作为人体细胞的“能量制造厂”，为细胞进行各种生命活动提供能量支持[1-2]。线粒体作为调控细胞能量代谢、生物合成及细胞衰亡的重要细胞器，其功能异常在许多疾病和病理状态的发生和发展过程中起着重要作用[3]。本研究通过被动免疫造模法建立ITP小鼠模型，以模型小鼠脾脏组织活性氧（Reactive Oxygen Species，ROS）含量、线粒体DNA（Mitochondrial DNA，mtDNA）相对拷贝数，以及结肠组织（重要免疫器官）ATP含量为观察指标，试图验证健脾益气摄血方提升缓解ITP小鼠乏力症状与改善线粒体功能的相关性。

1 材料与方法

1.1 材料……