LncRNA PVT1 靶向 miR-410-3p 调控高糖诱导的人肾小管上皮细胞中炎性和纤维化成分表达的实验研究

刘燕霞; 邵志远; 田宝金; 耿周

doi:10.13898/j.cnki.issn.2097-5252.2025.04.003

LncRNA PVT1 靶向 miR-410-3p 调控高糖诱导的人肾小管上皮细胞中炎性和纤维化成分表达的实验研究

Experimental study on LncRNA PVT1 targeting miR-410-3p to regulate the expression of inflammatory and fibrotic components in high glucose-induced human renal tubular epithelial cells

摘要

摘要:
目的探讨长链非编码RNA(LncRNA)PVT1是否靶向miR-410-3p调控高糖诱导的人肾小管上皮细胞中炎性和纤维化成分的表达。
方法将人肾小管上皮细胞HK-2分为空白对照(NC)组、高糖诱导(HG)组、si-NC+HG组、si-LncRNA PVT1+HG组、miR-NC+HG组、miR-410-3p+HG组、anti-miR-NC+si-LncRNA PVT1+HG组、anti-miR-410-3p+si-LncRNA PVT1+HG组。CCK-8法检测细胞活性，逆转录-定量聚合酶链反应检测肾小管上皮细胞HK-2的LncRNA PVT1和miR-410-3p表达情况，Western blotting检测纤维化因子α-平滑肌肌动蛋白(α-SMA)、纤连蛋白(Fn)、Ⅰ型胶原蛋白α1链(COL1α1)蛋白的表达，酶联免疫吸附测定法检测炎性因子肿瘤坏死因子-α(TNF-α)、白细胞介素-6(IL-6)的表达。双荧光素酶活性检测实验分析LncRNA PVT1与miR-410-3p的相互作用。
结果 HG组肾小管上皮细胞HK-2中LncRNA PVT1表达量明显高于NC组(P＜0.01)，miR-410-3p表达量和细胞活性较NC组明显降低(P＜0.01)，α-SMA、Fn、COL1α1蛋白表达量、TNF-α、IL-6含量均较NC组升高(P＜0.05)。抑制LncRNA PVT1表达或过表达miR-410-3p后，人肾小管上皮细胞HK-2活性升高(P＜0.05)，α-SMA、Fn、COL1α1蛋白表达量、TNF-α、IL-6含量降低(P＜0.05)。LncRNA PVT1靶向调控miR-410-3p的表达。敲低miR-410-3p表达可减弱抑制LncRNA PVT1表达对人肾小管上皮细胞HK-2中α-SMA、Fn、COL1α1蛋白表达、TNF-α、IL-6含量的影响(P＜0.01)。
结论抑制LncRNA PVT1表达通过靶向miR-410-3p，抑制高糖诱导的人肾小管上皮细胞中炎性和纤维化成分表达。

Abstract:
Objective To investigate whether long non-coding RNA(LncRNA) PVT1 targeting miR-410-3p to regulate the expression of inflammatory and fibrotic components in high glucose-induced human renal tubular epithelial cells.
Methods Human renal tubular epithelial cells HK-2 were divided into the blank control(NC) group, high glucose induced(HG) group, si-NC+HG group, si-LncRNA PVT1+HG group, miR-NC+HG group, miR-410-3p+HG group, anti-miR-NC+si-LncRNA PVT1+HG group and anti-miR-410-3p+si-LncRNA PVT1+HG group.The cell activity was detected by CCK-8 assay.The mRNA expression levels of LncRNA PVT1 and miR-410-3p in renal tubular epithelial cells HK-2 were detected by reverse transcription-quantitative polymerase chain reaction, the protein expression levels of fibrosis factor α-smooth muscle actin(α-SMA), fibronectin(Fn), collagen type Ⅰ α1 chain(COL1a1) were detected by Western blotting, and the contents of tumor necrosis factor-α(TNF-α) and interleukin-6(IL-6) were detected by enzyme-linked immunosorbent assay.The interaction between LncRNA PVT1 and miR-410-3p was analyzed by dual luciferase activity assay. Results: The expression level of LncRNA PVT1 in renal tubular epithelial cells HK-2 in the HG group was significantly higher than that in NC group(P < 0.01), the expression level of miR-410-3p and cell activity in the HG group were significantly lower than that in NC group(P < 0.01), and the protein expression levels of α-SMA, Fn, COL1a1, and contents of TNF-α, IL-6 in the HG group were significantly higher than those in the NC group(P < 0.05).After inhibiting the expression of LncRNA PVT1 or overexpressing miR-410-3p, the cell activity of in renal tubular epithelial cells HK-2 increased(P < 0.05), and the protein expression levels of α-SMA, Fn, COL1a1, and contents of TNF-α, IL-6 decreased(P < 0.05).The LncRNA PVT1 targetedly regulated the expression of miR-410-3p.Down-regulating the expression of miR-410-3p could attenuate the inhibitory effects of LncRNA PVT1 expression on α-SMA, Fn, COL1a1 protein expression, TNF-α and IL-6 content in human renal tubular epithelial cells HK-2(P < 0.01).
Conclusions Inhibition of LncRNA PVT1 expression inhibits the expression of inflammatory and fibrotic components in high glucose-induced human renal tubular epithelial cells by targeting miR-410-3p.

HTML全文

参考文献(29)

施引文献

资源附件(0)