长链非编码 RNA TCF7 通过与 miR-200c 形成海绵样结构触发内质网应激在糖尿病肾病患者中。
LncRNA TCF7 triggered endoplasmic reticulum stress through a sponge action with miR-200c in patients with diabetic nephropathy.
机构信息
Department of Nephrology, Central Hospital of Jiangjin District, Chongqing, China.
出版信息
Eur Rev Med Pharmacol Sci. 2019 Jul;23(13):5912-5922. doi: 10.26355/eurrev_201907_18336.
OBJECTIVE
To investigate the functions and mechanisms of long non-coding RNA TCF7 (LncTCF7) in patients with diabetic nephropathy (DN).
PATIENTS AND METHODS
LncTCF7 and miR-200c expressions in DN (+) were detected by Real Time-Polymerase Chain Reaction (RT-PCR). In the established high-glucose group (HG) model, RT-PCR and Western blot were used to detect the expressions of lncTCF7 and key apoptotic genes. The effect of podocyte endoplasmic reticulum (ER) stress was detected in high glucose conditions after lncTCF7 siRNA transfection. The miR-200c level was detected in lncTCF7 overexpressing cell model and the luciferase reporter gene assay was performed to verify the potential binding site of lncTCF7 with miR-200c. Furthermore, ER stress-associated genes were detected in patients with DN. Finally, in the HG model, the levels of ER stress were detected by WB after transfection with miR-200c inhibitor and lncTCF7 siRNA with miR-200c mimics.
RESULTS
Results showed that the lncTCF7 expression was up-regulated, while miR-200c was significantly downregulated in patients with DN (+). In the HG model, the expression of lncTCF7 was significantly increased and some key ER stress-associated genes, such as CHOP, XBP1, and cleaved caspase3, were significantly increased, while the anti-apoptotic protein Bcl-2 was significantly decreased. However, after inhibiting the lncTCF7 expression, these gene levels were reversed. In lncTCF7 overexpressing cells, miR-200c expression was significantly down-regulated compared with the control (p<0.05) and the luciferase reporter gene assay results showed that lncTCF7 could directly bind to miR-200c. In the HG model, after inhibiting lncTCF7 expression, the miR-200c level was increased, while the ER stress-associated proteins CHOP, XBP1, and cleaved caspase3 were significantly repressed. However, these proteins were reversed after inhibiting miR-200c expression. In addition, the expressions of ER stress-associated protein and apoptotic protein in human DN patients were consistent with HG cell model.
CONCLUSIONS
LncRNA TCF7 triggered endoplasmic reticulum stress through a sponge action with miR-200c in patients with diabetic nephropathy.
目的
研究长链非编码 RNA TCF7（LncTCF7）在糖尿病肾病（DN）患者中的功能和作用机制。
方法
采用实时聚合酶链反应（RT-PCR）检测 DN（+）患者中 LncTCF7 和 miR-200c 的表达。在建立高葡萄糖组（HG）模型后，采用 RT-PCR 和 Western blot 检测 lncTCF7 和关键凋亡基因的表达。在高葡萄糖条件下，通过 LncTCF7 siRNA 转染检测足细胞内质网（ER）应激的影响。在 LncTCF7 过表达细胞模型中检测 miR-200c 水平，并通过荧光素酶报告基因检测验证 LncTCF7 与 miR-200c 的潜在结合位点。此外，检测 DN 患者中与 ER 应激相关的基因。最后，在 HG 模型中，转染 miR-200c 抑制剂和 LncTCF7 siRNA 后，通过 WB 检测 ER 应激水平，并转染 miR-200c 模拟物。
结果
结果表明，DN（+）患者的 LncTCF7 表达上调，而 miR-200c 表达显著下调。在 HG 模型中，LncTCF7 的表达显著增加，一些关键的 ER 应激相关基因，如 CHOP、XBP1 和 cleaved caspase3，表达显著增加，而抗凋亡蛋白 Bcl-2 则显著减少。然而，抑制 LncTCF7 的表达后，这些基因水平得到逆转。在 LncTCF7 过表达细胞中，与对照组相比，miR-200c 的表达明显降低（p<0.05），荧光素酶报告基因检测结果表明 LncTCF7 可以直接与 miR-200c 结合。在 HG 模型中，抑制 LncTCF7 表达后，miR-200c 水平升高，而 ER 应激相关蛋白 CHOP、XBP1 和 cleaved caspase3 明显受到抑制。然而，抑制 miR-200c 表达后，这些蛋白被逆转。此外，DN 患者中 ER 应激相关蛋白和凋亡蛋白的表达与 HG 细胞模型一致。
结论
在糖尿病肾病患者中，LncRNA TCF7 通过与 miR-200c 的海绵作用引发内质网应激。
Zotero 插件