| Objective:The aim of the work is to study the characteristics of mitochondrial quality control, oxidavtive state and inflammatory cytokines expression in mouse skeletal muscle under fifferent exercise protocol in order to find the relationship between ROS, mitochondrial qaulity control and IL-6 generation. Explore Nrf-2 and its effects on different signal pathways and the regulation mechanism of IL-6 biological effectiveness, in order to find the role of ROS and mitochondrial homeostasis in IL-6 anti-inflammatory effects.Methods:Male C57BL/6 mice were performed a continuous concentric (CE) or ecconcentric exercise(DE) for 1 hour. Time sequential analysis related indies at sedentary state (control), exercise state (E30, E60), and convalescence state (PE30, PE60, PE3h and PE6h) respectively. Another part of mice in the same state were partitioned dividied into concentric or ecconcentric exercise, and each of them were further dividied into four group:control group, drench group, exercise group and drech and exercise jointed group. All trans retinoic acidand Sulforaphane were emploryed in concentric and ecconcentric exercise group respectively. Animals were trained and killed on the 4th day after 3 days drench. HE stains of mice skeletal muscle slide to explore morphology and inflammatory cell infiltration. Mitochondrial respiratory functions, member potential and ROS generation ratio were detected from fresh mitochondril which were extracted from mice skeletal muscle. Oxidative stress indies were determined from skeletal muscle homogenates. mRNA and protein levels of mitochondrial biogenesis/dynamic gene, and inf lammton related gene were measured by Realtime PCR and Westernblot respectively.Results:1. Muscle-derived IL-6 presented an impulse-type expression in CE group but sustained increase with inflammatory cell infiltration and TNF-a content increase in serum in DE group. Mitochondrial ROS generation rate is consistent with the trend in two groups at different time points.2. PGC-1 a expression in skeletal muscle dramatically increase started from E30 point in CE group-Compared with Control group, DE p65 expression was significantly increased at each time points, and TNF-α presented a gradual increase from PE60 to final in DE group.3. Mitochondrial functions were strengthened with PGC-1α, COX Ⅳ,OPA1 and MFN2 lever increased during E30 to PE60 comparied with Control in CE group. Compared to Control group, Drpl and Fisl level were increased from E60 to PE60 point in DE group.4. After up-regulated Nrf-2 expression, mitochondrial functions were strengthened with mitochondrial ROS generation ratio and antioxidant enzyme activity increased. Meantime, PGC-1α, COX Ⅳ and IL-6 expression increased, but p65 and TNF-α expressions decrased in skeletal muscle.Conclusions:1. Different exercise can active different signal pathway to induce muscle-derived IL-6 expression, whichi preseanted the same change trend with mitochondrial ROS generation suggested the signal role of mitochondrial drived ROS.2. A about of concentric exercise can up-regulate PGC-1α expression to mobilize mitochondria and accelerate antioxidase expression, promote mitochondrial biogenesis and fusion process, contribute to delay the exercise-induced TNF-α increase, which were suggested PGC-1α can suppressed NF-κB and related inflammation response, in order to participated in biological effects of IL-6 regulation by mitochondrial homeostasis.3. Nrf-2 can activate PGC-1α expression, further influence mitochondrial function and antioxidant capacity, which were suggested PGC-1α may inhibit oxidative injury of ROS and NF-κB expression to suppression inflammation response, and involved the anti-inflammation effect of IL-6 in skeletal muscle. |
PDF Full Text Request