Phytochemical investigation of the decoction of Urtica fissa rhizomes led to the isolation of 23 known compounds. Their structures were identified as medioresinol dimethyl ether (1), L-pyroglutamic acid methyl ester (2), nicotinic acid (3), L-pyroglutamic acid (4), erythritol (5), 6-methyl-2'-deoxy thymidine (6), 2-methyl-6-(2',3',4'-trihydroxybutyl)-pyrazine (7), 5-hydroxyl-2-hydroxymethyl pyridine (8), adenine (9), uracil (10), thymine (11), adenosine (12), inosine (13), 2'-deoxyadenosine (14), 2'-deoxyguanosine (15), 2'-deoxyinosine (16), uridine (17), n-butyl-O-β-D-fructopyranoside (18), di-D-fructose (19), β-D-fructofuranosyl- α-D-galactopyranoside (20), his (5-formyl-furfuryl) ether (21), chlorogenic acid (22), and 5-hydroxymethyl furaldehyde (23) by spectroscopic methods. In addition, a total of 20 compounds (1-20) were isolated from U. fissa for the first time. Meanwhile, compounds 1, 6, 7, 8, 19 and 20 were isolated from the Urticaceae plants for the first time.