Lu Jing, Miao Jiangang, Sun Jianhua
Department of Nephropathy, Yucheng People's Hospital, Yucheng, Shandong 251200, P.R. China.
Department of CT, Yucheng People's Hospital, Yucheng, Shandong 251200, P.R. China.
Exp Ther Med. 2020 Apr;19(4):2833-2840. doi: 10.3892/etm.2020.8534. Epub 2020 Feb 18.
Long noncoding (Lnc)RNA np_5318 has been proved to be involved in renal injury, while its functionality in renal ischemia-reperfusion (I/R) injury is unknown. Therefore, the present study aimed to investigate the role of lncRNA np_5318 in the development of renal I/R injury. Renal I/R injury model and I/R cell model were established . The expression of np_5318 in I/R cell was inhibited by small interfering (si)-np_5318 and increased by pc-np_5318. Renal function was detected and evaluated by automatic biochemical tests. Immunohistochemical staining was performed to detect the expression cluster of differentiation (CD)31, transforming growth factor (TGF)-β1 and (mothers against decapentaplegic homolog 3) Smad3 in renal tissue. The interaction between np_5318 and Smad3 was verified by chromatin immunoprecipitation (ChIP). Western blotting was performed to detect the expression levels of TGF-β1, Smad3 and phosphorylated (p)-Smad3 in renal tissue and renal cells. Expression of np_5318 in renal tissue and renal cells was detected by reverse transcription-quantitative PCR. Relative cell viability was confirmed by MTT assay. Renal function was impaired and pathological changes in renal tissue were observed in the renal I/R injury group, indicating the renal I/R injury model was successfully established. Compared with the sham group, the expression level of np_5318 significantly increased in the renal I/R injury group. ChIP data confirmed the interaction between np_5318 and Smad3. The expression of TGF-β1, Smad3 and p-Smad3 in renal tissue was also significantly increased in the renal I/R injury group. Furthermore, the I/R cell model was successfully constructed and np_5318 in I/R group was significantly increased compared with the control group. Cell growth was significantly suppressed in the I/R group compared with the control group. Additionally, transfection with pc-np_5318 significantly inhibited cell growth of I/R cells at 48 and 72 h. While inhibition of np_5318 by si-np_5318 significantly increased the cell growth of I/R cells at 48 and 72 h. Moreover, the level of TGF-β1, p-Smad3 and Smad3 was significantly increased in the I/R group compared with the control group, and transfection with pc-np_5318 significantly increased the level of TGF-β1, p-Smad3 and Smad3. While inhibition of np_5318 by si-np_5318 significantly suppressed the level of TGF-β1, p-Smad3 and Smad3. LncRNA np_5318 may participate in the development of renal I/R injury through TGF-β/Smad signaling pathway.
长链非编码（Lnc）RNA np_5318已被证明与肾损伤有关，但其在肾缺血再灌注（I/R）损伤中的功能尚不清楚。因此，本研究旨在探讨lncRNA np_5318在肾I/R损伤发生发展中的作用。建立了肾I/R损伤模型和I/R细胞模型。小干扰（si）-np_5318抑制I/R细胞中np_5318的表达，而pc-np_5318则使其表达增加。通过自动生化检测来检测和评估肾功能。进行免疫组织化学染色以检测肾组织中分化簇（CD）31、转化生长因子（TGF）-β1和（抗五聚体蛋白同源物3）Smad3的表达。通过染色质免疫沉淀（ChIP）验证np_5318与Smad3之间的相互作用。采用蛋白质免疫印迹法检测肾组织和肾细胞中TGF-β1、Smad3和磷酸化（p）-Smad3的表达水平。通过逆转录定量PCR检测肾组织和肾细胞中np_5318的表达。通过MTT法确认相对细胞活力。肾I/R损伤组肾功能受损，肾组织出现病理变化，表明成功建立了肾I/R损伤模型。与假手术组相比，肾I/R损伤组中np_5318的表达水平显著升高。ChIP数据证实了np_5318与Smad3之间的相互作用。肾I/R损伤组肾组织中TGF-β1、Smad3和p-Smad3的表达也显著增加。此外，成功构建了I/R细胞模型，与对照组相比，I/R组中np_5318显著增加。与对照组相比，I/R组细胞生长明显受到抑制。此外，在48和72小时时，pc-np_5318转染显著抑制I/R细胞的生长。而si-np_5318抑制np_5318在I/R细胞48和72小时时显著增加细胞生长。此外，与对照组相比，I/R组中TGF-β1、p-Smad3和Smad3水平显著升高，pc-np_5318转染显著增加TGF-β1、p-Smad3和Smad3水平。而si-np_5318抑制np_5318显著抑制TGF-β1、p-Smad3和Smad3水平。LncRNA np_5318可能通过TGF-β/Smad信号通路参与肾I/R损伤的发生发展。
Zotero 插件
