Canadine from Corydalis turtschaninovii Stimulates Myoblast Differentiation and Protects against Myotube Atrophy

Abstract

Cachexia and sarcopenia are the main causes of muscle atrophy. These result in a reduction in the muscle fiber area, myo-protein content, and muscle strength, with various molecular modulators being involved. Although several reports have proposed potential therapeutic agents, no effective treatments have been found for muscle atrophy. We searched for myogenic modulators from medicinal plants to treat muscle diseases. We isolated six alkaloids from Corydalis turtschaninovii and evaluated their myogenic potential by using the MyoD reporter gene assay in C2C12 cells. Among the tested compounds, canadine showed the strongest transactivation of MyoD and increased MHC expression during myogenesis. The activation of p38 MAP kinase and Akt are major mechanisms that contribute to the myogenesis by canadine. Canadine increased the number of multinucleated and cylinder-shaped myotubes during myogenesis of C2C12 myoblasts. To determine the preventive effect of canadine in cancer-induced muscle wasting, differentiated C2C12 myotubes were treated with conditioned media from CT26 colon carcinoma culture (CT26 CM) in the presence of canadine. Canadine ameliorated the muscle protein degradation caused by CT26-CM by down-regulating the muscle specific-E3 ligases, MAFbx/atrogin-1 and MuRF1. In this study, we found that canadine from C. turtschaninovii stimulates myogenesis and also inhibits muscle protein degradation. Therefore, we suggest canadine as a protective agent against muscle atrophy.

A Chalcone from Ashitaba (Angelica keiskei) Stimulates Myoblast Differentiation and Inhibits Dexamethasone-Induced Muscle Atrophy

“…Mouse skeletal myoblast C2C12 is derived from mouse satellite cell and well-established cell line to study the myogenic potential of chemicals [21]. C2C12 myoblasts were differentiated with EAK (10, 100, or 1000 ng/mL), which dose-dependently enhanced expression levels of MHC (Figure 2A,B) as a marker of terminal myogenesis [19]. We observed the cylinder-shaped multinucleated myotubes (containing more than four nuclei and indicating mature myotubes) by immunostaining for MHC and DAPI [19].…”

“…C2C12 myoblasts were differentiated with EAK (10, 100, or 1000 ng/mL), which dose-dependently enhanced expression levels of MHC (Figure 2A,B) as a marker of terminal myogenesis [19]. We observed the cylinder-shaped multinucleated myotubes (containing more than four nuclei and indicating mature myotubes) by immunostaining for MHC and DAPI [19]. The number of multinucleated myotubes was increased by EAK in a dose-dependent manner.…”

“…C2C12 myoblast cells obtained from the American Type Culture Collection (ATCC, Manassas, VA, USA) were differentiated for 3 days as demonstrated in our previous studies [19]. We prepared CT26 murine colon carcinoma cell-conditioned medium (CM) according to previous studies [19]. To prepare 4-HD-treated CM (HD-CM), CT26 cells were plated and treated with 4-HD for 24 h in DMEM containing 10% FBS.…”

“…To measure MyoD transcriptional activity, C2C12 cells were transiently transfected with MyoD-responsive reporter 4RTK-luciferase (RTK-Luc) and pBP-MyoD constructs using Lipofectamine ® 2000 Reagent (Invitrogen, Carlsbad, CA, USA) [19]. At 24 h post transfection, cells were treated with test compounds for 24 h. Luciferase assay was then performed using cell lysates and a commercial luciferase assay kit (Promega, Madison, WI, USA).…”

Molecular mechanisms of cancer cachexia‑induced muscle atrophy (Review)

“…a number of proinflammatory and transfer factors, in addition to the activation of several pathways have been identified in skeletal muscle and were illustrated to be involved in cancer cachexia-induced muscle atrophy; for example, TnF-α (47), Twist1 (48), the nF-κB signaling pathway (49) and the p38 MaPK signaling pathway (45), which were all discovered to be upregulated. The overexpression of proinflammatory factors, transfer factors or members of signaling pathways in skeletal muscle in the context of cancer cachexia eventually converge on the MurF1 and MaFbx of the uPS, promoting proteasome hydrolysis in the uPS and leading to skeletal muscle protein degradation (50)(51)(52)(53).…”