摘要
低氧可能对机体代谢异常有着积极的调节作用。Apelin是G蛋白偶联受体APJ的内源性配体,参与骨骼肌能量代谢的调节。目的:采用动物实验,观察低氧暴露不同时间对小鼠骨骼肌apelin表达的影响;通过细胞实验进一步探讨HIF-1α是否参与了低氧对小鼠骨骼肌apelin表达的调节。方法:动物实验采用C57BL/6J雄性小鼠40只,随机分为对照组(0 h)和不同时间低氧暴露组(6 h、24 h、48 h),每组10只,共4组。低氧组氧浓度约为11. 2%,低氧暴露结束后即刻取材。细胞实验采用C2C12细胞,分为常氧对照组(N)、低氧暴露组(H)、低氧对照组(H+siControl)和低氧HIF-1α沉默组(H+siHIF-1α),共4组。转染48 h沉默后,在低氧浓度为5%的低氧中培养72 h,收集细胞。Western blot测定骨骼肌组织和细胞apelin、APJ、HIF-1α的蛋白表达。RT-PCR测定骨骼肌中apelin、APJ、HIF-1αmRNA含量。结果:1)与0 h组相比,6 h、24 h和48 h组小鼠骨骼肌apelin/APJ mRNA和蛋白及HIF-1αmRNA表达均增加;与6 h组相比,48 h组小鼠骨骼肌apelin mRNA及HIF-1αmRNA和蛋白表达量增加;与24 h组相比,48 h组小鼠骨骼肌apelin和HIF-1αmRNA表达量显著增加。2)与N组相比,H组细胞apelin、APJ、HIF-1α蛋白表达增加;与H组和H+siControl组相比,H+siHIF-1α组细胞apelin、APJ、HIF-1α蛋白表达降低。结论:模拟常压低氧暴露(11. 2%氧浓度) 6 h可显著增加小鼠骨骼肌apelin/APJ的表达,并且其表达水平呈时间依赖性;HIF-1α参与了低氧对骨骼肌apelin/APJ表达的调节作用。揭示了低氧与骨骼肌apelin表达的关系,为低氧暴露促进健康提供理论依据。
Hypoxia may have a positive role in the regulation of metabolic abnormalities. Apelin is the endogenous ligand for G protein-coupled receptor APJ,it can adjust the energy metabolism in skeletal muscle. Objectives: To investigate the effect of different duration of hypoxia on the expression of apelin/APJ in skeletal muscle of mice by animal experiment;and to detect whether HIF-1α is involved in the regulation of apelin expression in mice skeletal muscle by hypoxia through cell experiment. Methods: In animal experiment,40 male C57 BL/6j mice were divided into control group (0 h) and different duration hypoxia groups (6 h,24 h,48 h) randomly. There were 10 mice in each group and 4 groups in total. The oxygen concentration in hypoxia group was about 11. 2%. The mice were sacrificed instantly after hypoxic exposure. In cell experiment,C2 C12 cells were allocated into four groups:normoxia group (N),hypoxic exposure group (H),hypoxic exposure control group (H + siControl) and hypoxic exposure with HIF-1α silence group (H + siHIF-1α). After 48 h transfection,the cells were cultured for 72 h at 5% oxygen concentration and then were collected. The protein expressions of apelin,APJ and HIF-1α in skeletal muscle tissue and cells were tested by Western blot;and the mRNA expressions of apelin,APJ,HIF-1α mRNA were measured by RT-PCR. Results: 1) Compared with 0 h group,the expressions of apelin/APJ and HIF-1αmRNA in skeletal muscle in 6 h,24 h and 48 h groups increased. Compared with the 6 h group,the expressions of apelin mRNA and HIF-1α in 48 h group increased significantly. Compared with 24 h group,the expression of apelin and HIF-1α mRNA in 48 h group increased significantly. 2) Compared with N group,the expression of apelin,APJ,HIF-1α protein in H group increased. Compared with H and H + siControl groups,the protein expression of apelin,APJ,HIF-1α in H + siHIF-1α group decreased. Conclusions: The 6 h simulated normal pressure hypoxia exposure (11. 2% oxygen concentration) could increase the expression of apelin/
作者
姬卫秀
张缨
JI Wei-xiu;ZHANG Ying(Beijing Sport University,Beijing 100084,China;Tianjin University of Sport,Tianjin 301617,China)
出处
《北京体育大学学报》
CSSCI
北大核心
2018年第12期73-78,共6页
Journal of Beijing Sport University
基金
国家自然科学基金资助项目(编号:31640044)
中央高校基础科研业务费专项基金资助项目(编号:2016GJ001)
