The purpose of this study was to analyze the muscle fatigue for lower limbs during the level and downhill running. The subjects were 6 males of twenties who have no experience to get the injury in the lower limbs and required to run on the level and downhill which was -7% grade treadmill at 8.3km/h. EMG signal was gained by ME3000P8 Measurement Unit and computed the Median Frequency(MF) with the power spectrum analysis in the Megawin software. Rectus femoris(RF), Vastus lateralis(VL), Gluteus medius(GLU), Biceps Femoris(BF), gastrocnemius medial head(GM), gastrocnemius lateral head(GL), Tibialis anterior(TA) were selected. The result of this study were as follows: The MF of RF decreased in the downhill running than level running in length of time but, the MF of VL was opposite. The MF of BF decreased in the level and downhill running, but, the MF of BF decreased much in the level than downhill running. The MF of GLU decreased much in the downhill running but, almost no change in the level running. The MF of TA decreased in the level running than downhill running. The MF of GL decreased in the level running but, the MF of GM decreased in the downhill running in length of time. This study analyzed the muscle fatigue of the lower limbs with the median frequency on the basis of an assumption that the impact force for the flexion and extension of the joint and the body mass may be much in the eccentric contraction such as the downhill running than level running. RF and GM showed the muscle fatigue in the downhill running than level running. BF and GL showed the muscle fatigue in the level running than downhill running.