| 引用本文: |
李腾辉,董一昕,闫凤娜,王淑艳,王珊珊,于雪,王旭,许明阳,李卫红.补中益气汤对PC12细胞线粒体DNA部分缺失模型的保护作用[J].湖南中医药大学学报,2020,40(12):1473-1478[点击复制] |
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| 补中益气汤对PC12细胞线粒体DNA部分缺失模型的保护作用 |
| 李腾辉,董一昕,闫凤娜,王淑艳,王珊珊,于雪,王旭,许明阳,李卫红 |
| (北京中医药大学中医学院, 北京 100029;首都医科大学附属北京地坛医院, 北京 100020) |
| 摘要: |
| 目的 探讨补中益气汤对PC12细胞线粒体DNA（mtDNA）部分缺失模型的保护作用及其分子机制。方法 利用神经细胞株PC12细胞，溴化乙锭诱导法制备mtDNA部分缺失模型。将PC12细胞分为4组：正常组、模型组、达纳康组、补中益气汤组。给药24 h后，CCK-8法检测各组细胞线粒体活性；比色法检测线粒体呼吸链复合物Ⅰ、Ⅱ、Ⅲ、Ⅳ活性及ATP含量；RT-PCR法检测各组线粒体转录因子A（mitochondrial transcription factor A，TFAM）表达。结果 500 ng/mL浓度溴化乙锭能够使细胞mtDNA拷贝数下降（P<0.01）造成PC12细胞mtDNA部分缺失。与正常组相比，模型组细胞线粒体活性和呼吸链复合物Ⅰ-Ⅳ活性降低（P<0.05，P<0.01），ATP含量减少（P<0.01），TFAM基因表达量下降（P<0.01）。与模型组相比，补中益气汤组线粒体活性显著升高（P<0.01），呼吸链复合物Ⅰ、Ⅱ、Ⅳ活性有所升高（P<0.05，P<0.01），ATP含量和TFAM基因表达量增加（P<0.01），治疗效果与达纳康组无统计学差异（P>0.05）。结论 补中益气汤对溴化乙锭所致mtDNA部分缺失模型具有保护作用，可显著改善线粒体的代谢功能，其机制可能与上调TFAM表达有关，这可能是补中益气汤发挥"补气"功能的现代生物学基础。 |
| 关键词: 补中益气汤 PC12细胞 线粒体 DNA部分缺失 |
| DOI：10.3969/j.issn.1674-070X.2020.12.007 |
| 投稿时间：2020-05-21 |
| 基金项目:北京中医药大学新奥奖励基金课题（2017-XAJLJJ-002）。 |
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| Protective Effect of Buzhong Yiqi Decoction on Partial Deletion Models of Mitochondrial DNA in PC12 Cells |
| LI Tenghui,DONG Yixin,YAN Fengna,WANG Shuyan,WANG Shanshan,YU Xue,WANG Xu,XU Mingyang,LI Weihong |
| (School of Traditional Chinese Medicine, Beijing University of Chinese Medicine, Beijing 100029, China;Beijing Ditan Hospital, Capital Medical University, Beijing 100020, China) |
| Abstract: |
| Objective To explore the protective effect of Buzhong Yiqi Decoction on partial deletion models of mitochondrial DNA (mtDNA) in PC12 cells and its molecular mechanism. Methods By using a nerve cell strain of PC12 cells, the ethidium bromide induction method was used to prepare partial deletion models of mtDNA. The PC12 cells were divided into 4 groups:a normal group, a model group, a Tanakan group and a Buzhong Yiqi Decoction group. Twenty-four hours after administration, the CCK-8 method was used to detect mitochondrial activities of the cells in each group; colorimetry was used to detect activities of mitochondrial respiratory chain complexes Ⅰ, Ⅱ, Ⅲ, Ⅳ and ATP contents; and the RT-PCR method was used to detect expressions of mitochondrial transcription factor A (TFAM) in each group. Results Ethidium bromide at a concentration of 500 ng/mL could decrease the copy number of cell mitochondrial DNA (P<0.01), resulting in partial deletion of mitochondrial DNA in the PC12 cells. Compared with the normal group, the mitochondrial activity and activities of respiratory chain complexes I-IV in the model group were decreased (P<0.05, P<0.01), the ATP content was decreased (P<0.01), and the TFAM gene expression was decreased. Compared with the model group, the mitochondrial activity in the Buzhong Yiqi Decoction group was significantly increased (P<0.01), the activities of respiratory chain complexes I, II and IV were increased (P<0.05, P<0.01), the content of ATP and the expression of TFAM gene were increased (P<0.01), and the efficacy was not statistically different from that in the Tanakan group (P>0.05). Conclusion Buzhong Yiqi Decoction has a protective effect on the partial deletion models of mitochondrial DNA induced by ethidium bromide, which can significantly improve the mitochondrial metabolic function. The mechanism may be related to up-regulation of the TFAM expression, which may be a modern biological basis of Buzhong Yiqi Decoction exerting the function of supplementing Qi. |
| Key words: Buzhong Yiqi Decoction PC12 cells mitochondria DNA partial deletion model |
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