脾气虚大鼠肝脏细胞线粒体蛋白质控的研究

Study on Mitochondrial Protein Control of Liver Cells in Spleen Qi Deficiency Rats

目的:观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。

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.