左旋肉碱通过诱导 deltex E3 泛素连接酶 3L 负向调节 Runx2/COL1A1 轴,缓解与恶病质相关的骨骼肌纤维化。

IF 9.4 1区 医学 Q1 GERIATRICS & GERONTOLOGY
Zongliang Lu, Li Wang, Zhenyu Huo, Na Li, Ning Tong, Feifei Chong, Jie Liu, Yaowen Zhang, Hongxia Xu
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RUNX family transcription factor 2 (Runx2), a transcription factor, and collagen type I alpha 1 chain (COL1A1), the principal constituent of SMF, have been linked previously, with Runx2 shown to directly modulate COL1A1 mRNA levels. <span>l</span>-Carnitine, a marker of cancer cachexia, can alleviate fibrosis in liver and kidney models; however, its role in cancer cachexia-associated fibrosis and the involvement of Runx2 in the process remain unexplored.</p>\n </section>\n \n <section>\n \n <h3> Methods</h3>\n \n <p>Female C57 mice (48 weeks old) were inoculated subcutaneously with MC38 cells to establish a cancer cachexia model. A 5 mg/kg dose of <span>l</span>-carnitine or an equivalent volume of water was administered for 14 days via oral gavage, followed by assessments of muscle function (grip strength) and fibrosis. 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引用次数: 0

摘要

背景:癌症恶病质诱发的骨骼肌纤维化(SMF)会影响肌肉再生、改变肌肉结构和功能、降低抗癌药物的疗效、降低患者的生活质量并缩短总生存期。RUNX家族转录因子2(Runx2)是一种转录因子,它与SMF的主要成分I型胶原蛋白α1链(COL1A1)有关联,Runx2可直接调节COL1A1 mRNA水平:雌性 C57 小鼠(48 周大)皮下接种 MC38 细胞,建立癌症恶病质模型。连续14天通过口服灌胃给药5毫克/千克剂量的左旋肉碱或同等体积的水,然后评估肌肉功能(握力)和纤维化。为了阐明Deltex E3泛素连接酶3L(DTX3L)/Runx2/COL1A1轴与转化生长因子β1刺激的NIH/3T3细胞纤维化之间的相互作用,研究人员使用了一系列分子技术,包括定量实时PCR、Western印迹分析、共免疫沉淀、分子对接、免疫荧光和Duolink检测。结果:补充左旋肉碱可减少癌症恶病质诱发的握力下降(>88.2%,P 57.9%,P 结论:该研究揭示了一种以前未被认识的新的腓肠肌肌酸激酶--DTX3L/Runx2/COL1A1轴:这项研究揭示了Runx2和DTX3L在SMF中的联系,并证明了左旋肉碱对癌症恶病质相关SMF有显著的治疗作用,这可能是通过上调DTX3L实现的。
本文章由计算机程序翻译,如有差异,请以英文原文为准。

l-Carnitine relieves cachexia-related skeletal muscle fibrosis by inducing deltex E3 ubiquitin ligase 3L to negatively regulate the Runx2/COL1A1 axis

l-Carnitine relieves cachexia-related skeletal muscle fibrosis by inducing deltex E3 ubiquitin ligase 3L to negatively regulate the Runx2/COL1A1 axis

Background

Cancer cachexia-induced skeletal muscle fibrosis (SMF) impairs muscle regeneration, alters the muscle structure and function, reduces the efficacy of anticancer drugs, diminishes the patient's quality of life and shortens overall survival. RUNX family transcription factor 2 (Runx2), a transcription factor, and collagen type I alpha 1 chain (COL1A1), the principal constituent of SMF, have been linked previously, with Runx2 shown to directly modulate COL1A1 mRNA levels. l-Carnitine, a marker of cancer cachexia, can alleviate fibrosis in liver and kidney models; however, its role in cancer cachexia-associated fibrosis and the involvement of Runx2 in the process remain unexplored.

Methods

Female C57 mice (48 weeks old) were inoculated subcutaneously with MC38 cells to establish a cancer cachexia model. A 5 mg/kg dose of l-carnitine or an equivalent volume of water was administered for 14 days via oral gavage, followed by assessments of muscle function (grip strength) and fibrosis. To elucidate the interplay between the deltex E3 ubiquitin ligase 3L(DTX3L)/Runx2/COL1A1 axis and fibrosis in transforming growth factor beta 1-stimulated NIH/3T3 cells, a suite of molecular techniques, including quantitative real-time PCR, western blot analysis, co-immunoprecipitation, molecular docking, immunofluorescence and Duolink assays, were used. The relevance of the DTX3L/Runx2/COL1A1 axis in the gastrocnemius was also explored in the in vivo model.

Results

l-Carnitine supplementation reduced cancer cachexia-induced declines in grip strength (>88.2%, P < 0.05) and the collagen fibre area within the gastrocnemius (>57.9%, P < 0.05). At the 5 mg/kg dose, l-carnitine also suppressed COL1A1 and alpha-smooth muscle actin (α-SMA) protein expression, which are markers of SMF and myofibroblasts. Analyses of the TRRUST database indicated that Runx2 regulates both COL1A1 and COL1A2. In vitro, l-carnitine diminished Runx2 protein levels and promoted its ubiquitination. Overexpression of Runx2 abolished the effects of l-carnitine on COL1A1 and α-SMA. Co-immunoprecipitation, molecular docking, immunofluorescence and Duolink assays confirmed an interaction between DTX3L and Runx2, with l-carnitine enhancing this interaction to promote Runx2 ubiquitination. l-Carnitine supplementation restored DTX3L levels to those observed under non-cachectic conditions, both in vitro and in vivo. Knockdown of DTX3L abolished the effects of l-carnitine on Runx2, COL1A1 and α-SMA in vitro. The expression of DTX3L was negatively correlated with the levels of Runx2 and COL1A1 in untreated NIH/3T3 cells.

Conclusions

This study revealed a previously unrecognized link between Runx2 and DTX3L in SMF and demonstrated that l-carnitine exerted a significant therapeutic impact on cancer cachexia-associated SMF, potentially through the upregulation of DTX3L.

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来源期刊
Journal of Cachexia Sarcopenia and Muscle
Journal of Cachexia Sarcopenia and Muscle MEDICINE, GENERAL & INTERNAL-
CiteScore
13.30
自引率
12.40%
发文量
234
审稿时长
16 weeks
期刊介绍: The Journal of Cachexia, Sarcopenia and Muscle is a peer-reviewed international journal dedicated to publishing materials related to cachexia and sarcopenia, as well as body composition and its physiological and pathophysiological changes across the lifespan and in response to various illnesses from all fields of life sciences. The journal aims to provide a reliable resource for professionals interested in related research or involved in the clinical care of affected patients, such as those suffering from AIDS, cancer, chronic heart failure, chronic lung disease, liver cirrhosis, chronic kidney failure, rheumatoid arthritis, or sepsis.
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