Wang Yun, Xu Yi, Zhao Tun, Ma Ya-Jun, Qin Wei, Hu Wen-Li
Department of Neurology, Beijing Chaoyang Hospital, Capital Medical University, Beijing 100020, China.
Department of Pharmacy, First Affiliated Hospital of Wenzhou Medical University, Wenzhou, Zhejiang, 325000, China.
Open Life Sci. 2024 Sep 9;19(1):20220952. doi: 10.1515/biol-2022-0952. eCollection 2024.
Intensive care unit-acquired weakness (ICU-AW) is prevalent in critical care, with limited treatment options. Certain microRNAs, like miR-542, are highly expressed in ICU-AW patients. This study investigates the regulatory role and mechanisms of miR-542 in ICU-AW and explores the clinical potential of miR-542 inhibitors. ICU-AW models were established in C57BL/6 mice through cecal ligation and puncture (CLP) and in mouse C2C12 myoblasts through TNF-α treatment. experiments demonstrated decreased muscle strength, muscle fiber atrophy, widened intercellular spaces, and increased miR-542-3p/5p expression in ICU-AW mice model. experiments indicated suppressed ATG5, ATG7 and LC3II/I, elevated MDA and ROS levels, decreased SOD levels, and reduced MMP in the model group. Similar to animal experiments, the expression of miR-542-3p/5p was upregulated. Gel electrophoresis explored the binding of polyethyleneimine/mesoporous silica nanoparticles (PEI/MMNs) to locked nucleic acid (LNA) miR-542 inhibitor (LNA-542). PEI/MMNs@LNA-542 with positive charge (3.03 ± 0.363 mV) and narrow size (206.94 ± 6.19 nm) were characterized. Immunofluorescence indicated significant internalization with no apparent cytotoxicity. Biological activity, examined through intraperitoneal injection, showed that PEI/MMNs@LNA-542 alleviated muscle strength decline, restored fiber damage, and recovered mitochondrial injury in mice. In conclusion, PEI/MMNs nanoparticles effectively delivered LNA-542, targeting ATG5 to inhibit autophagy and alleviate mitochondrial damage, thereby improving ICU-AW.
重症监护病房获得性肌无力（ICU-AW）在重症监护中很常见，治疗选择有限。某些微小RNA，如miR-542，在ICU-AW患者中高表达。本研究探讨miR-542在ICU-AW中的调控作用及机制，并探索miR-542抑制剂的临床潜力。通过盲肠结扎和穿刺（CLP）在C57BL/6小鼠中建立ICU-AW模型，并通过TNF-α处理在小鼠C2C12成肌细胞中建立模型。实验表明，在ICU-AW小鼠模型中，肌肉力量下降、肌纤维萎缩、细胞间隙增宽以及miR-542-3p/5p表达增加。实验表明，模型组中自噬相关蛋白5（ATG5）、自噬相关蛋白7（ATG7）和微管相关蛋白1轻链3Ⅱ/Ⅰ（LC3II/I）受到抑制，丙二醛（MDA）和活性氧（ROS）水平升高，超氧化物歧化酶（SOD）水平降低，线粒体膜电位（MMP）降低。与动物实验类似，miR-542-3p/5p的表达上调。凝胶电泳探究了聚乙烯亚胺/介孔二氧化硅纳米颗粒（PEI/MMNs）与锁核酸（LNA）miR-542抑制剂（LNA-542）的结合情况。对带正电荷（3.03±0.363 mV）且尺寸较小（206.94±6.19 nm）的PEI/MMNs@LNA-542进行了表征。免疫荧光显示其有明显的内化且无明显细胞毒性。通过腹腔注射检测生物活性，结果显示PEI/MMNs@LNA-542可减轻小鼠肌肉力量下降，修复纤维损伤并恢复线粒体损伤。总之，PEI/MMNs纳米颗粒有效地递送了LNA-542，靶向ATG5以抑制自噬并减轻线粒体损伤，从而改善ICU-AW。
