Jellyfish envenomation is a common medical problem in many countries. However, the myotoxicity and effector molecules of scyphozoan venoms remain uninvestigated. Here, we present the myotoxicity of nematocyst venom from Nemopilema nomurai (NnNV), a giant venomous scyphozoan from China, for the first time, using in vivo models with inhibitors. NnNV was able to induce remarkable myotoxicity including significant muscle swelling, increasing the content of CK and LDH in serum, stimulating inflammation of muscle tissue, and destroying the structure of muscle tissue. In addition, the metalloproteinase inhibitors BMT and EDTA significantly reduced the myotoxicity induced by NnNV. Moreover, BMT and EDTA could decrease the inflammatory stimulation and necrosis of muscle tissue caused by the venom. These observations suggest that the metalloproteinase components of NnNV make a considerable contribution to myotoxicity. This study contributes to understanding the effector molecules of muscle injury caused by jellyfish stings and suggests a new idea for the treatment of scyphozoan envenomation.