中国骨质疏松杂志

基于“骨肉不相亲”探讨补肾健脾法对OP大鼠肌骨中ULK1/FUNDC1介导的线粒体自噬的影响

To investigate the effect of invigorating kidney and invigorating spleen on ULK1/FUNDC1 mediated mitochondrial autophagy in OP rats

投稿时间：2023-11-22 修订日期：2024-02-28

DOI：

中文关键词: 骨肉不相亲 ULK1/FUNDC1通路线粒体自噬骨质疏松

英文关键词:flesh and blood do not match ULK1/FUNDC1 channel Mitochondrial autophagy osteoporosis

基金项目:

作者	单位	邮编
付夜平	辽宁中医药大学	110847
杨芳^*	辽宁中医药大学	110847
孙鑫	辽宁中医药大学
李俊儒	辽宁中医药大学
王致远	辽宁中医药大学
杨蓝鑫	辽宁中医药大学
刘彤	辽宁中医药大学
张李博	辽宁中医药大学

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中文摘要:

摘要：目的：探究补肾健脾法对OP大鼠肌骨中ULK1/FUNDC1介导的线粒体自噬对的影响。方法：72只2月龄雄性SD大鼠随机分为正常组、模型组、补肾组、健脾组、补肾健脾组、西药组（福善美），每组12只，模型组及各个用药组采用尾部悬吊法进行造模，造模时间为5周。正常组与模型组予以超纯水灌胃，其余各组予以相应药物进行灌胃。造模结束后行腹主动脉采血，酶联免疫吸附法（ELISA）测定血清碱性磷酸酶（ALP）、抗酒石酸酸性磷酸酶（TRACP）含量；计算机微断层扫描（Micro-CT）检测大鼠左侧股骨远端形态学变化；双能X线行股骨骨密度（BMD）检测；马松（Masson）染色观察腓肠肌病理变化；蛋白免疫印迹法（Western blot）检测股骨、腓肠肌中线粒体蛋白ULK1、FUNDC1、LC3-Ⅱ/LC3-Ⅰ蛋白表达。结果：与正常组相较，模型组ALP（P＜0.01）降低、TRACP（P＜0.01）升高，骨小梁面积、BMD（P＜0.01）均下降，提示造模成功。Masson染色提示在中医不同治法干预下腓肠肌组织纤维化程度得到缓解。Western blot结果显示：在中医不同治法干预下与模型组股骨相比ULK1、LC3-Ⅱ/LC3-Ⅰ、FUNDC1表达降低（P＜0.05），线粒体自噬水平降低；与模型组腓肠肌相比ULK1、FUNDC1、LC3-Ⅱ/LC3-Ⅰ蛋白表达升高（P＜0.05），线粒体自噬水平升高。结论：补肾健脾中医治法对于OP模型大鼠的治疗效果可能是通过ULK1/FUNDC1通路调控线粒体自噬发挥作用，综合来看补肾健脾法效果较优。

英文摘要:

Abstract: Objective:To explore the effect of invigorating kidney and invigorating spleen on ULK1/FUNDC1 mediated mitochondrial autophagy in OP rats. Methods: 72 2-month-old male SD rats were randomly divided into normal group, model group, kidney-tonifying group, kidney-tonifying group, spleen-strengthening group and western medicine group (Fosamaxa), with 12 rats in each group. The model group and each medication group were modeled by tail suspension method, and the modeling time was 5 weeks. The normal group and model group were given ultra-pure water, and the other groups were given corresponding drugs. After the modeling, blood samples were collected from the aorta, and the serum alkaline phosphatase (ALP) and tartrate-resistant acid phosphatase (TRACP) were determined by enzyme-related immunosorbent assay (ELISA). The morphological changes of left femur were detected by Micro-CT. Bone mineral density (BMD) of left femur was detected by dual-energy X-ray. The pathological changes of left gastrocnemius were observed by Masson staining. The expressions of mitochondrial proteins ULK1, FUNDC1, LC3-Ⅱ/LC3-Ⅰ extracted from the left femur and gastrocnemius were detected by Western blot.Result:Compared with normal group, ALP (P < 0.01) decreased, TRACP (P < 0.01) increased, trabecular area and BMD (P < 0.01) decreased in model group, indicating successful modeling. Masson staining suggested that the degree of gastrocnemius fibrosis was relieved under the intervention of different treatment methods of traditional Chinese medicine. Western blot results showed that the expressions of ULK1, LC3-Ⅱ/LC3-Ⅰ and FUNDC1 were decreased (P < 0.05) and the level of mitochondrial autophagy was decreased compared with that of model femur group under different treatment methods of traditional Chinese medicine. Compared with model group, the protein expressions of ULK1, FUNDC1, LC3-Ⅱ/LC3-Ⅰ were increased (P < 0.05), and the level of mitochondrial autophagy was increased.Conclusion:The therapeutic effect of TCM treatment of tonifying kidney and spleen on OP model rats may be due to the regulation of mitochondrial autophagy through ULK1/FUNDC1 pathway, and the effect of tonifying kidney and spleen is better.

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