王艺璇 刘文俊 许欣竹 马丽娜 陈文娜 宋囡 单德红

【摘要】 目的：觀察肝细胞线粒体结构和功能及其蛋白质控，探讨肝耐受脾气虚的可能机制。方法：16只SPF级雄性SD大鼠根据随机数字表法平均分成两组，即8只正常组（对照组）和8只脾气虚组（模型组）。透射电镜观察肝细胞线粒体结构，比色法和JC-1法分别检测肝组织ATP和肝细胞线粒体膜电位（membrane membrane potential， MMP），Western blot法检测肝细胞线粒体热休克蛋白（hot shock protein， HSP）70、Lon蛋白酶（Lon protease， LonP）以及酪蛋白裂解酶（caseinolytic protease， Clp）XP复合物等表达。结果：两组线粒体大量分布于细胞核附近，形态相似，均呈卵圆形，边缘清楚，内部致密，未见到明显异常变化。与正常组相比，模型组ATP和MMP水平均下降，差异均有统计学意义（P<0.01）。与正常组相比，模型组HSP70、ClpX、ClpP表达均上升，LonP表达下降，差异均有统计学意义（P<0.01）。结论：肝耐受脾气虚的机制与肝细胞线粒体蛋白质控机制增强有关。

【关键词】 脾气虚 线粒体 HSP70蛋白 Lon蛋白酶 ClpXP复合物

Study on Mitochondrial Protein Control of Liver Cells in Spleen Qi Deficiency Rats/WANG Yixuan， LIU Wenjun， XU Xinzhu， MA Lina， CHEN Wenna， SONG Nan， SHAN Dehong. //Medical Innovation of China， 2021， 18（14）： 0-022

[Abstract] Objective： To observe the structure， function and protein control of hepatocyte mitochondria and explore the possible mechanism of liver tolerance to spleen qi deficiency. Method： A total of 16 SPF male SD rats were divided into two groups according to random number table method， namely 8 normal group （control group） and 8 spleen qi deficiency group （model group）. The mitochondrial structure of liver cells was observed by transmission electron microscopy. The ATP of liver tissue and the mitochondrial membrane potential （MMP） of liver cells were detected by colorimetric method and JC-1 method， respectively. Western blot was used to detect expressions of hot shock protein （HSP70）， Lon protease （LonP） and caseinolytic protease （Clp） XP complex. Result： A large number of mitochondria in the two groups were distributed near the nucleus， and the morphology was similar， with oval shape， clear edge and dense interior， and no obvious abnormal changes were observed. Compared with the normal group， the levels of ATP and MMP in the model group were decreased， the differences were statistically significant （P<0.01）. Compared with the normal group， the expressions of HSP70， ClpX and ClpP in the model group were all increased， while the expression of LonP was decreased， the differences were statistically significant （P<0.01）. Conclusion： The mechanism of liver tolerance to spleen qi deficiency is related to the enhancement of the quality control mechanism of mitochondrial protein in liver cells.

[Key words] Spleen qi deficiency Mitochondron Hot shock protein 70 protein Lon protease ClpXP complex

First-authors address： Graduate School， Liaoning University of Traditional Chinese Medicine， Shenyang 110847， China

doi：10.3969/j.issn.1674-4985.2021.14.005

传统中医理论认为肝木克脾土，特别是脾虚易为肝乘，即“土虚木乘”，还指出脾气虚能扰肝，如“脾气虚则四支不用，五脏不安”。由此可知脾气虚不能改变肝克脾的规律，或者说肝对脾气虚具有一定的耐受性，但其现代医学机制不明。中西医对肝的形态描述基本一致，如位于腹腔右胁内、右叶大和左叶小等。现代医学认为肝脏是物质代谢的关键部位，而线粒体不仅是肝细胞的动力工厂，还广泛参与生物合成、信号转导和胞内钙稳态维持等，因此其健康对于肝至关重要。线粒体DNA突变率高，常导致其参与编码的线粒体蛋白错误折叠，但热休克蛋白70（hot shock protein， HSP70）能够对其再折叠实现修复，对于那些不能够被修复的错误折叠蛋白会被呈递给Lon蛋……