Koyama Tomohisa, Uchida Kentaro, Itakura Makoto, Miyagi Masayuki, Tazawa Ryo, Inoue Gen, Fukushima Kensuke, Ohashi Yoshihisa, Tsukada Ayumi, Takaso Masashi
Department of Orthopedic Surgery, Kitasato University School of Medicine, 1-15-1 Minami-ku, Kitasato, Sagamihara City 252-0374, Kanagawa, Japan.
Department of Biochemistry, Kitasato University School of Medicine, 1-15-1 Minami-ku, Kitasato, Sagamihara City 252-0374, Kanagawa, Japan.
Curr Issues Mol Biol. 2022 Jul 8;44(7):3146-3155. doi: 10.3390/cimb44070217.
Expression of the apelin receptor, APJ, in skeletal muscle (SM) is known to decrease with age, but the underlying mechanism remains unclear. Increased tumor necrosis factor (TNF)-α levels are observed in SM with age and are associated with muscle atrophy. To investigate the possible interconnection between TNF-α elevation and APJ reduction with aging, we investigated the effect of TNF-α on APJ expression in cells derived from the quadriceps femoris of C57BL/6J mice. Expression of and in the quadriceps femoris was compared between 4- (young) and 24-month-old (old) C57BL/6J mice ( = 10 each) using qPCR. Additionally, APJ-positive cells and TNF-α protein were analyzed by flow cytometry and Western blotting, respectively. Further, quadricep-derived cells were exposed to 0 (control) or 25 ng/mL TNF-α, and the effect on expression was examined by qRT-PCR. expression and the ratio of APJ-positive cells among quadricep cells were significantly lower in old compared to young mice. In contrast, levels of mRNA and TNF-α protein were significantly elevated in old compared to young mice. Exposing young and old derived quadricep cells to TNF-α for 8 and 24 h caused levels to significantly decrease. TNF-α suppresses APJ expression in muscle cells in vitro. The increase in TNF-α observed in SM with age may induce a decrease in APJ expression.
已知阿片肽受体APJ在骨骼肌(SM)中的表达会随着年龄的增长而降低,但其潜在机制仍不清楚。随着年龄的增长,在骨骼肌中观察到肿瘤坏死因子(TNF)-α水平升高,且与肌肉萎缩有关。为了研究TNF-α升高与衰老过程中APJ减少之间可能存在的联系,我们研究了TNF-α对来自C57BL/6J小鼠股四头肌的细胞中APJ表达的影响。使用qPCR比较了4个月大(年轻)和24个月大(年老)的C57BL/6J小鼠(每组n = 10)股四头肌中APJ和TNF-α的表达。此外,分别通过流式细胞术和蛋白质免疫印迹法分析了APJ阳性细胞和TNF-α蛋白。进一步地,将股四头肌来源的细胞暴露于0(对照)或25 ng/mL的TNF-α中,并通过qRT-PCR检测其对APJ表达的影响。与年轻小鼠相比,年老小鼠股四头肌细胞中APJ的表达和APJ阳性细胞的比例显著降低。相反,与年轻小鼠相比,年老小鼠中TNF-α mRNA水平和TNF-α蛋白水平显著升高。将年轻和年老小鼠来源的股四头肌细胞暴露于TNF-α 8小时和24小时后,APJ水平显著降低。TNF-α在体外抑制肌肉细胞中APJ的表达。随着年龄增长在骨骼肌中观察到的TNF-α增加可能会导致APJ表达下降。