Col18A1反义lncRNA在肾纤维化过程中的多重保护作用及机制

At early stage of renal fibrosis, autophagy helps to reduce cell oxidative stress, extracellular matrix deposition and inflammation, and fibrosis. Therefore, for the treatment and prognosis of renal fibrosis, it is of important significance to study the regulation mechanism of autophagy. Long non-coding RNA (lncRNA) can act as a competitive endogenous RNA (ceRNA) sponging multiple miRNA to indirectly regulate, but also directly regulate expression of key genes related to autophagy and fibrosis at the transcriptional and post transcriptional levels. Our group recently found that miR-21 and miR-34a were able to target multiple key genes related to autophagy and fibrosis, inhibited autophagy at early stage of renal fibrosis and exacerbated the degree of renal fibrosis. Besides, miR-21 and miR-34a could bind and interact with Col18A1 antisense (AS) lncRNA. However, the role of Col18A1 AS lncRNA in autophagy during early renal fibrosis is not clear. This study aims to explore the role of Col18A1 AS lncRNA on autophagy and the progression of renal fibrosis in vivo and in vitro. Rat model of renal ischemia reperfusion injury (IRI) induced chronic renal interstitial fibrosis is to be established, and primary rat renal tubular epithelial cells are to be cultured. Using the luciferase report gene, RNA pull-down, and RNase protection assay, we are to investigate the interaction of Col18A1 AS lncRNA with miR-21, miR-34a and Col18A1 mRNA respectively, and to clarify the multiple protective effects and mechanisms of Col18A1 AS lncRNA in the progression of renal fibrosis. The findings will provide theoretical basis for the autophagy-based treatment of renal fibrosis.

肾纤维化早期，自噬有助于减轻细胞氧化应激、炎症反应及纤维化程度，因此深入研究自噬调控机制对肾纤维化的治疗有重要意义。长链非编码RNA（lncRNA）既可结合多个miRNA间接调控，亦可转录和转录后水平直接调控自噬和纤维化关键基因。本团队前期发现miR-21和miR-34a可靶定多个自噬和纤维化相关基因，抑制肾纤维化自噬，加剧纤维化程度，且miR-21/34a均与Col18A1 AS lncRNA相互作用。而Col18A1 AS lncRNA在肾纤维化自噬中的作用尚不清楚。本研究拟构建肾缺血再灌注损伤大鼠模型，并分离培养原代肾小管上皮细胞，在体内外水平探索其对自噬和肾纤维化进程的影响；并运用荧光素酶报告基因、RNA pull-down、核酸酶保护分析等技术研究其与miR-21/34a和Col18A1 mRNA的相互作用，阐明其在肾纤维过程中的多重保护作用及机制，为肾纤维化治疗提供理论基础。

肾纤维化早期，自噬有助于减轻细胞氧化应激、炎症反应及纤维化程度，因此深入研.究自噬调控机制对肾纤维化的治疗有重要意义。长链非编码RNA（lncRNA）既可结合多个m.iRNA间接调控，亦可转录和转录后水平直接调控自噬和纤维化关键基因。本团队前期发现.miR-21和miR-34a可靶定多个自噬和纤维化相关基因，抑制肾纤维化自噬，加剧纤维化程.度，且miR-21/34a均与Col18A1 AS lncRNA相互作用。而Col18A1 AS lncRNA在肾纤维化自.噬中的作用尚不清楚。本研究拟构建肾缺血再灌注损伤大鼠模型，并分离培养原代肾小管.上皮细胞，在体内外水平探索其对自噬和肾纤维化进程的影响；并运用荧光素酶报告基因.、RNA pull-down、核酸酶保护分析等技术研究其与miR-21/34a和Col18A1 mRNA的相互作.用，阐明其在肾纤维过程中的多重保护作用及机制，为肾纤维化治疗提供理论基础。