Investigation into the role of Stmn2 in vascular smooth muscle phenotype transformation during vascular injury via RNA sequencing and experimental validation
Investigation into the role of Stmn2 in vascular smooth muscle phenotype transformation during vascular injury via RNA sequencing and experimental validation
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DOI：
10.1007/s11356-021-15846-7
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发表时间：
2021-08
影响因子：
5.8
通讯作者：
Xiao Ke;W. Guo;Yanren Peng;Zongming Feng;Yi-teng Huang;Ming Deng;M. Wei;Zanxin Wang
中科院分区：
文献类型：
--
作者：
Xiao Ke;W. Guo;Yanren Peng;Zongming Feng;Yi-teng Huang;Ming Deng;M. Wei;Zanxin Wang
本研究通过RNA测序和实验验证，研究了Stmn2在血管损伤中对血管平滑肌表型转化的影响。分别于伤后1、3、5d提取总RNA进行RNA测序，筛选差异表达基因。Western印迹法检测Stmn2及其相关靶点的蛋白表达。苏木精-伊红(H&E)染色观察大鼠颈动脉的形态变化。免疫组织化学方法检测血管平滑肌细胞表型标志物--平滑肌α-肌动蛋白(α-SMA)、波形蛋白和骨桥蛋白的表达。DEGS与细胞外基质和质膜外的其他细胞成分有关。它们与蛋白质结合、细胞骨架蛋白结合、信号受体结合等分子功能、肌动蛋白细胞骨架调控等京都百科全书中的基因和基因组途径相关。Stmn2被认为是肌动蛋白细胞骨架途径与血管疾病的中枢基因，在血管损伤过程中其表达呈先降低后升高的趋势。Western印迹分析显示，血管损伤后即刻Stmn2和微管蛋白表达下降，Stmn2过表达显著上调骨桥蛋白、α-SMA和Vimentin的表达。形态分析和免疫组织化学染色结果还显示，Stmn2过表达促进损伤血管组织内膜增厚，促进增殖细胞核抗原表达。综上所述，我们的结果提示Stmn2在血管损伤中可能起到潜在的作用，这可能与VSMC的表型转化有关。需要进一步研究以确定Stmn2在血管损伤中的详细分子机制。
This study examined the effects of Stmn2 on phenotype transformation of vascular smooth muscle in vascular injury via RNA sequencing and experimental validation. Total RNA was extracted for RNA sequencing after 1, 3 and 5 days of injury to screen the differentially expressed genes (DEGs). Western blot was used to detect the protein expression of Stmn2 and its associated targets. The morphological changes of carotid arteries in rats were examined by hematoxylin and eosin (H&E) staining. The expression of vascular smooth muscle cell (VSMC) phenotype markers smooth muscle alpha-actin (α-SMA), vimentin and OPN were detected by immunohistochemistry. DEGs were related to the extracellular matrix and other cell components outside the plasma membrane. They were associated with protein binding, cytoskeleton protein binding, signal receptor binding and other molecular functions, actin cytoskeleton regulation and other Kyoto Encyclopedia of Genes and Genomes pathways. Stmn2 was identified as the hub gene of actin cytoskeleton pathway and vascular disease, and its expression followed the trend of decreasing initially and increasing afterwards during the progress of vascular injury. Western blot assay showed that the expression of Stmn2 and Tubulin decreased immediately after vascular injury; Stmn2 overexpression significantly up-regulated the expression of osteopontin and α-SMA and vimentin in VSMCs. The results of morphology analysis and immunostaining also showed that Stmn2 overexpression promoted the intima thickening and enhanced the proliferating cell nuclear antigen expression in the injured vascular tissues. In conclusion, our results implied that Stmn2 may play a potential role in vascular injury, which may be associated with VSMC phenotype transformation. Further studies are warranted to determine detailed molecular mechanisms of Stmn2 in vascular injury.