PDF(Pubmed)
Abstract：
Introduction: Carnosol exhibited ameliorating effects on muscle atrophy of mice developed cancer cachexia in our previous research. Method: Here, the ameliorating effects of carnosol on the C2C12 myotube atrophy result from simulated cancer cachexia injury, the conditioned medium of the C26 tumor cells or the LLC tumor cells, were observed. To clarify the mechanisms of carnosol, the possible direct target proteins of carnosol were searched using DARTS (drug affinity responsive target stability) assay and then confirmed using CETSA (cellular thermal shift assay). Furthermore, proteomic analysis was used to search its possible indirect target proteins by comparing the protein expression profiles of C2C12 myotubes under treatment of C26 medium, with or without the presence of carnosol. The signal network between the direct and indirect target proteins of carnosol was then constructed. Results: Our results showed that, Delta-1-pyrroline-5-carboxylate synthase (P5CS) might be the direct target protein of carnosol in myotubes. The influence of carnosol on amino acid metabolism downstream of P5CS was confirmed. Carnosol could upregulate the expression of proteins related to glutathione metabolism, anti-oxidant system, and heat shock response. Knockdown of P5CS could also ameliorate myotube atrophy and further enhance the ameliorating effects of carnosol. Discussion: These results suggested that carnosol might ameliorate cancer cachexia-associated myotube atrophy by targeting P5CS and its downstream pathways.
摘要：
简介：在我们先前的研究中，Carnosol对发展为癌症恶病质的小鼠的肌肉萎缩具有改善作用。方法：在这里，肌醇对C2C12肌管萎缩的改善作用是由模拟癌症恶病质损伤引起的，C26肿瘤细胞或LLC肿瘤细胞的条件培养基，被观察到。为了阐明鼠尾草酚的机制，使用DARTS（药物亲和力响应靶标稳定性）测定搜索了鼠尾草酚的可能直接靶蛋白，然后使用CETSA（细胞热转移测定）进行了确认。此外，蛋白质组学分析用于通过比较C26培养基处理下C2C12肌管的蛋白质表达谱来搜索其可能的间接靶蛋白，有或没有鼠尾草的存在。然后构建了鼠尾草酚直接和间接靶蛋白之间的信号网络。结果:我们的研究结果表明,Delta-1-吡咯啉-5-羧酸合酶（P5CS）可能是肌管中鼠尾草的直接靶蛋白。证实了鼠尾草酚对P5CS下游氨基酸代谢的影响。鼠尾草酚可上调谷胱甘肽代谢相关蛋白的表达,抗氧化系统，和热休克反应。击倒P5CS还可以改善肌管萎缩，并进一步增强鼠尾草的改善作用。讨论：这些结果表明，鼠尾草酚可能通过靶向P5CS及其下游途径改善与恶病质相关的肌管萎缩。
