Kim Minjee, Chun Jaemoo, Jung Hyun Ah, Choi Jae Sue, Kim Yeong Shik
Natural Products Research Institute, College of Pharmacy, Seoul National University, Seoul 08826, Republic of Korea.
Department of Food Science and Human Nutrition, Chonbuk National University, Jeonju 54896, Republic of Korea.
Phytomedicine. 2017 Aug 15;32:30-36. doi: 10.1016/j.phymed.2017.04.007. Epub 2017 Apr 13.
Intense exercise has the potential to increase oxidative stress and cause muscle damage. Mitogen-activated protein kinases (MAPKs) and nuclear factor-κB (NF-κB) are two major regulators of gene transcription in response to oxidative stress in the skeletal muscle. Pure capillarisin (CAP) isolated from Artemisia capillaris Thunberg is known to have antioxidant and anti-inflammatory effects.
HYPOTHESIS/PURPOSE: We hypothesized CAP to exert antioxidant activity against exercise-induced oxidative stress and suppress acute inflammatory response. We aimed to investigate skeletal muscle recovery after intense exercise with or without CAP administration.
Eccentric exercise was conducted to induce muscle damage (C57BL6 mice, 13m/min for 60min downhill running). Mice were divided into four groups (n=6): the rested control, exercised, and exercised with CAP treatments (20mg/kg and 80mg/kg, ip injection 24h prior to exercise) groups.
After the intense exercise, mice were sacrificed immediately, and after 24h the gastrocnemius muscles and blood plasma were collected for further study. The DCFH-DA and TBARS assays were conducted for anti-oxidative capacity. Muscle damage markers, creatinine phosphate kinase (CPK) and lactate dehydrogenase (LDH) were investigated at plasma level. Muscle data were examined with H&E staining and microscopy. MAPK and NF-κB pathway, chemokine and cytokine productions were confirmed by western blotting and RT-PCR.
From DCFH-DA and TBARS assays, exercise increased the level of ROS production, but these changes were suppressed by CAP treatment. Exercise induced muscle damage by raising the levels of soluble muscle enzymes, such as CPK and LDH. However, this result was improved in CAP-treated groups at plasma level. Exercise activated MAPK (ERK 1/2 and JNK but not p38) and NF-κB (nuclear p50 and p65, and cytosolic p-IκBα) subunits at protein level but CAP attenuated these increase in a dose dependent manner. At the mRNA level, the chemokines CINC-1 and MCP-1, and cytokine IL-6 in gastrocnemius muscle were increased by exercise, whereas CAP suppressed these increase.
Overall, our results indicate that CAP, as a single compound, can attenuate muscle damage by exerting antioxidant and anti-inflammatory effects. Thus, CAP is a potential candidate for the muscle protective agent in the future.
高强度运动有可能增加氧化应激并导致肌肉损伤。丝裂原活化蛋白激酶(MAPKs)和核因子-κB(NF-κB)是骨骼肌中响应氧化应激的基因转录的两个主要调节因子。已知从茵陈蒿中分离出的纯茵陈色原酮(CAP)具有抗氧化和抗炎作用。
假设/目的:我们假设CAP对运动诱导的氧化应激具有抗氧化活性,并抑制急性炎症反应。我们旨在研究在给予或不给予CAP的情况下,高强度运动后骨骼肌的恢复情况。
进行离心运动以诱导肌肉损伤(C57BL6小鼠,以13米/分钟的速度下坡跑60分钟)。将小鼠分为四组(n = 6):静息对照组、运动组以及运动并给予CAP处理组(20毫克/千克和80毫克/千克,在运动前24小时腹腔注射)。
高强度运动后,立即处死小鼠,并在24小时后收集腓肠肌和血浆用于进一步研究。进行DCFH-DA和TBARS测定以评估抗氧化能力。在血浆水平检测肌肉损伤标志物磷酸肌酸激酶(CPK)和乳酸脱氢酶(LDH)。用苏木精-伊红染色和显微镜检查肌肉数据。通过蛋白质印迹法和逆转录-聚合酶链反应确认MAPK和NF-κB途径、趋化因子和细胞因子的产生。
从DCFH-DA和TBARS测定结果来看,运动增加了活性氧的产生水平,但这些变化被CAP处理所抑制。运动通过提高可溶性肌肉酶如CPK和LDH的水平诱导肌肉损伤。然而,在血浆水平,CAP处理组的这一结果得到改善。运动在蛋白质水平激活了MAPK(细胞外信号调节激酶1/2和应激活化蛋白激酶,但不包括p38)和NF-κB(核p50和p65以及胞质磷酸化IκBα)亚基,但CAP以剂量依赖的方式减弱了这些增加。在mRNA水平,运动使腓肠肌中的趋化因子CINC-1和MCP-1以及细胞因子白细胞介素-6增加,而CAP抑制了这些增加。
总体而言,我们的结果表明,CAP作为单一化合物,可以通过发挥抗氧化和抗炎作用减轻肌肉损伤。因此,CAP是未来肌肉保护剂的潜在候选物。