Abstract

Diabetic nephropathy (DN) is one of the severe complications of diabetes. Nowadays, effective treatment for end-stage renal disease (ESRD) patients is still limited. HK-2 cells were stimulated with serum from phosphate-buffered saline (PBS) or Jiawei Shuilu Erxiandan (JSE)-treated DN mice, then long non-coding RNA (lncRNA) CLYBL-AS2 was discovered by RNA sequence, following the comparison of the serum from normal patients with DN patients to confirm the role of lncCLYBL-AS2. Next, we performed in vitro studies to explore the effect of lncCLYBL-AS2 in DN and its molecular mechanism. Coptis, as one of the components of JSE, could decrease the expression of lncCLYBL-AS2, which is increased in DN and correlated with the severity of DN. Knockdown/overexpression of lncCLYBL-AS2 inhibited/promoted the invasion and fibrogenesis of HK-2 cells. Furthermore, lncCLYBL-AS2 was negatively correlated with miR-204-4p with a positive correlation with SNAI1; eventually, CLYBL-AS2 regulated SNAI1 by binding to miR-204-5p, which accounted for the inhibition of epithelial–mesenchymal transition (EMT) and fibrogenesis. LncCLYBL-AS2 inhibited by Coptis improved EMT and fibrogenesis in HK-2 cells through miR-204-5p-SNAI1 axis, therefore, lncCLYBL-AS2 could serve as a potential diagnosis and therapeutic target for DN.

摘要

糖尿病肾病（DN）是糖尿病的严重并发症之一。目前，终末期肾病（ESRD）患者的有效治疗仍然有限。用磷酸盐缓冲液 (PBS) 或加味水路二仙丹 (JSE) 处理的 DN 小鼠的血清刺激 HK-2 细胞，然后通过 RNA 序列比较发现长链非编码 RNA (lncRNA) CLYBL-AS2。正常患者和DN患者的血清以确认lncCLYBL-AS2的作用。接下来，我们进行了体外探讨lncCLYBL-AS2在DN中的作用及其分子机制的研究。黄连作为 JSE 的组成成分之一，可以降低 lncCLYBL-AS2 的表达，lncCLYBL-AS2 在 DN 中增加，并与 DN 的严重程度相关。lncCLYBL-AS2 的敲低/过表达抑制/促进了 HK-2 细胞的侵袭和纤维化。此外，lncCLYBL-AS2 与 miR-204-4p 呈负相关，与 SNAI1 呈正相关；最终，CLYBL-AS2 通过与 miR-204-5p 结合来调节 SNAI1，从而抑制上皮间质转化 (EMT) 和纤维化。黄连抑制的 LncCLYBL-AS2 通过 miR-204-5p-SNAI1 轴改善 HK-2 细胞的 EMT 和纤维化，因此，lncCLYBL-AS2 可作为 DN 的潜在诊断和治疗靶点。