基于miRNA-ox-LDL-SIRT1在自噬中的对话机制探讨益肾活血法干预慢性肾脏病的作用机理

Shen Kangling with the effect of Yi Shen Huo Xue is a traditional Chinese medicine in the treatment of pediatric nephropathy in Fujian Province, which is well-known and makes good effect in clinical practice from Wang Zhuchu,but the mechanism is not very clear. The latest research shows that miR-21/-29 is involved in the pathogenesis of CKD, and ox-LDL is the most representative lipoprotein in CKD process, and SIRT1 is necessary for autophagy, and plays a powerful renal protective role in CKD. Therefore, we propose the hypothesis that regulating the autophagy pathway of miRNA,ox-LDL,SIRT1 is the mechanism of Shenkangling in the treatment of CKD. This topic with SIRT1 -/- mice model of chronic kidney disease (CKD) and renal tubular epithelial cells as the research object, using the small animal vivo imaging technology, electron microscope, flow cytometry instrument and cellular molecular biology methods to observe the interaction mechanisms of mir-21/-29,ox-LDL,SIRT1 and their effects on autophagy in the course of CKD, to analyze the role of mir-21/-29,ox-LDL,SIRT1 and autophagy in the treatment of CKD with Shen Kangling and clarify the targets of Shenkangling. The successful completion of this project will provide theoretical basis for the clinical application of Shen Kangling in CKD and for improving the curative effect of CKD.

益肾活血肾康灵是福建省名老中医王著础治疗肾病临床经验方(院内制剂)，临床疗效显著，但作用机理仍不清楚。最新研究表明，miR-21/-29共同参与CKD的发病机制，ox-LDL是影响CKD进程最具代表性的脂蛋白，SIRT1为自噬所必需，且在CKD中发挥强大的肾脏保护作用。故我们提出科学假说：通过自噬途径调节miRNA-ox-LDL-SIRT1是肾康灵治疗CKD的作用机理。本课题以SIRT1-/-小鼠CKD模型和肾小管上皮细胞为研究对象，运用小动物活体成像、电镜、流式细胞仪及分子生物学方法，研究在CKD发病过程中，miR-21/-29、ox-LDL、SIRT1的对话机制及其对自噬的影响；分析miR-21/-29、ox-LDL、SIRT1及自噬在肾康灵治疗CKD肾损伤时的角色并明确肾康灵的作用靶点。本项目的顺利完成将为肾康灵在CKD中的临床应用提供理论依据，为提高CKD的疗效奠定一定的理论基础。

益肾活血中药肾康灵是福建省名老中医治疗肾病临床经验方，临床疗效显著，但作用机理仍不清楚。本研究以SIRT1基因敲除小鼠慢性肾脏病（CKD）模型和肾小管上皮细胞为研究对象，运用小动物活体成像、电镜、流式细胞仪及分子生物学方法，研究在CKD发病过程中，miR-21/-29、ox-LDL、SIRT1的对话机制及其对自噬的影响；分析miR-21/-29、ox-LDL、SIRT1及自噬在肾康灵治疗CKD肾损伤时的角色并明确肾康灵的作用靶点，以期为肾康灵在CKD中的临床应用提供理论依据，为提高CKD的疗效奠定一定的理论基础。本研究结果表明，肾康灵能够通过SIRT1调控自噬，从而改善腺嘌呤诱导的CKD小鼠肾功能和肾组织病理形态变化，对CKD小鼠肾损伤起保护作用； miR-21a-3p和miR-29c-3p能够靶向调控SIRT1基因，从而参与肾小管上皮细胞的自噬和凋亡损伤；ox-LDL可诱导肾小管上皮细胞发生凋亡损伤和自噬，肾康灵可能通过调控miR-21a-3p/miR-29c-3p及SIRT1介导的细胞自噬，抑制细胞凋亡损伤，促进肾小管上皮细胞增殖。