右归丸对肾阳虚证大鼠肾脏线粒体功能的影响

Effects of Yougui Pills on the Function of Kidney Mitochondria in Rats with Kidney-yang Deficiency Syndrome

目的观察右归丸对肾阳虚证大鼠肾脏线粒体功能的影响,探讨其调节能量代谢的作用机制。方法30只SD大鼠随机分为正常组(6只)和造模组(24只),造模组后肢肌注氢化可的松注射液,正常组注射2 mL生理盐水。成模后将造模组随机分为模型组和右归丸高、中、低剂量组,每组6只。右归丸高、中、低剂量组给予相应剂量汤剂灌胃,正常组和模型组给予生理盐水灌胃。透射电镜观察肾脏线粒体超微结构,试剂盒检测线粒体三磷酸腺苷(ATP)含量,JC-1荧光探针检测线粒体膜电位,DCFH-DA法检测细胞内活性氧(ROS)水平。结果与正常组比较,模型组大鼠肾脏线粒体明显肿胀,部分出现空泡,线粒体嵴排列不规则,基质不均匀,核膜不明显,线粒体ATP含量减少、膜电位下降,细胞内ROS水平升高,差异有统计学意义(P<0.01);与模型组比较,右归丸各剂量组大鼠肾脏线粒体结构均有改善,线粒体肿胀减轻,线粒体嵴排列较规则,基质均匀,核膜清晰,线粒体ATP含量增加、膜电位升高,细胞内ROS水平降低,其中右归丸高剂量组作用明显,差异有统计学意义(P<0.01)。结论右归丸通过改善大鼠线粒体功能调节能量代谢异常。

Objective To observe the effects of Yougui Pills on the function of kidney mitochondria in rats with kidney-yang deficiency;To explore its mechanism of action of energy metabolism.Methods Thirty SD rats were randomly divided into normal group(6 rats)and model group(24 rats).The model group was injected with hydrocortisone injection into the hindlimb muscle,while the normal group was injected with 2 mL normal saline.After the successful modeling,they were divided into the model group,Yougui Pills high-,medium-,and low-dosage groups,with 6 rats in each group.Yougui Pills high-,medium-,and low-dosage groups were given corresponding dosages of decoction for gavage,and the normal group and model group were given normal saline for gavage.The ultrastructure of kiney mitochondria was observed with transmission electron microscope;the kit was used to detect the mitochondrial adenosine triphosphate(ATP)content;the JC-1 fluorescent probe was used to detect the mitochondrial membrane potential;the DCFH-DA method was used to detect the intracellular reactive oxygen species(ROS)level.Results Compared with the normal group,the mitochondria of the kidneys of the model group were obviously swollen,some vacuoles appeared,the mitochondrial cristae were arranged irregularly,the matrix was uneven,and the nuclear membrane was not obvious;mitochondrial ATP content decreased significantly,membrane potential decreased,and intracellular ROS level increased,with statistical significance(P<0.01).Compared with the model group,the mitochondrial structure of kidney rats in Yougui Pills groups was improved,the swelling of mitochondria was reduced,the mitochondrial cristae arrangement was more regular,the matrix was uniform,and the nuclear membrane was clear;rat mitochondrial ATP content increased,membrane potential increased,and intracellular ROS level decreased.Among them,Yougui Pills high-dosage group had a significant effect,with statistical significance(P<0.01).Conclusion Yougui Pills can improve rat mitochondrial function and regulate abnormal energy metabolism.