基于 RNA 测序预测 lncRNA 在肾脏衰老中的作用。
Predict the role of lncRNA in kidney aging based on RNA sequencing.
机构信息
Dialysis Department of Nephrology Hospital, The First Affiliated Hospital of Xi'an Jiaotong University, West Yanta Road 277, 710061, Xi'an, Shaanxi, China.
Department of Nephrology, The First People's Hospital Lanzhou City, No1 Wujiayuan, 730050, Qilihe, Lanzhou, Gansu, China.
出版信息
BMC Genomics. 2022 Apr 2;23(1):254. doi: 10.1186/s12864-022-08479-8.
BACKGROUND
Long noncoding RNAs (lncRNAs) are involved in physiological and pathological processes. However, no studies have been conducted on the relationship between lncRNAs and renal aging.
RESULTS
First, we evaluated the histopathology of young (3-month-old) and old (24-month-old) C57BL/6J mouse kidneys. Masson trichrome staining and PAS staining showed interstitial collagen deposition and fibrosis, mesangial matrix expansion, a thicker basement membrane and renal interstitial fibrosis in old mouse kidneys. Senescence-associated β-galactosidase (SA-β-gal)-positive areas in the kidneys of old mice were significantly elevated compared to those of young mice. Then, we analyzed the differential expression of lncRNAs and mRNAs in the kidneys of young and old mouse kidneys by RNA-seq analysis. 42 known and 179 novel differentially expressed lncRNAs and 702 differential mRNAs were detected in the mouse kidney. Next, we focused on the differentially expressed mRNAs and lncRNAs by RNA-seq. GO and KEGG analyses were performed based on differentially expressed mRNAs between young and old mouse kidneys. Transregulation based on RIsearch and the correlation coefficient of mRNA-lncRNA were also calculated. The mRNA-lncRNA network was constructed by choosing a Spearman correlation coefficient > 0.9 or <-0.9. GO and KEGG pathway enrichment analyses revealed that differentially expressed mRNAs participated in aging-related pathways. A total of 10 lncRNAs and trans-regulated mRNAs were constructed. Finally, we validated the role of lncRNA Gm43360 by CCK-8, flow cytometry, western blot and SA-β-gal staining. The expression level of Adra1a was positively correlated and Csnk1a1 was negatively correlated with lncRNA Gm43360. The cell counting kit-8 (CCK-8) results showed that lncRNA Gm43360 promoted cell viability. LncRNA Gm43360 increased the percentage of S phase cells and decreased the percentage of G1 phase cells compared with the negative control. LncRNA Gm43360 decreased the expression of p53, p21 and SA-β-gal.
CONCLUSIONS
LncRNA Gm43360 may play a protective role in kidney aging.
背景
长链非编码 RNA（lncRNA）参与生理和病理过程。然而，尚未有研究探讨 lncRNA 与肾脏衰老之间的关系。
结果
首先，我们评估了年轻（3 月龄）和年老（24 月龄）C57BL/6J 小鼠肾脏的组织病理学变化。Masson 三色染色和 PAS 染色显示，年老小鼠肾脏的间质胶原沉积和纤维化、系膜基质扩张、基底膜增厚和肾间质纤维化更为明显。与年轻小鼠相比，年老小鼠肾脏中衰老相关β-半乳糖苷酶（SA-β-gal）阳性区域显著增加。然后，我们通过 RNA-seq 分析检测了年轻和年老小鼠肾脏中 lncRNA 和 mRNA 的差异表达。在小鼠肾脏中检测到 42 个已知和 179 个新的差异表达 lncRNA 和 702 个差异表达 mRNA。接下来，我们通过 RNA-seq 关注差异表达的 mRNAs 和 lncRNAs。基于年轻和年老小鼠肾脏之间差异表达的 mRNAs 进行了 GO 和 KEGG 分析。还根据 RIsearch 和 mRNA-lncRNA 的相关系数计算了反式调控。通过选择 Spearman 相关系数 > 0.9 或 <-0.9 构建了 mRNA-lncRNA 网络。GO 和 KEGG 途径富集分析显示，差异表达的 mRNAs 参与了与衰老相关的途径。构建了总共 10 个 lncRNA 和反式调控的 mRNA。最后，我们通过 CCK-8、流式细胞术、western blot 和 SA-β-gal 染色验证了 lncRNA Gm43360 的作用。Adra1a 的表达水平与 lncRNA Gm43360 呈正相关，Csnk1a1 与 lncRNA Gm43360 呈负相关。细胞计数试剂盒-8（CCK-8）结果表明，lncRNA Gm43360 促进了细胞活力。与阴性对照相比，lncRNA Gm43360 增加了 S 期细胞的百分比，减少了 G1 期细胞的百分比。lncRNA Gm43360 降低了 p53、p21 和 SA-β-gal 的表达。
结论
lncRNA Gm43360 可能在肾脏衰老中发挥保护作用。
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