The invention belongs to the technical field of civil engineering, and specifically relates to a prestressing anti-fatigue girder. The prestressing anti-fatigue girder comprises a top flange, a bottom flange, a web, a bearer stiffening plate and prestressing anchor slab, anchor holes, a high-strength prestressing steel strand, an anchoring stiffening plate, an anchor head, an arc plate, a bearer steel bracket and lateral movement proof cushion block. The prestressing steel strand is penetrated into anchor holes which are near to a tensile flange and are symmetrical on two sides of the web, and is anchored by the anchor head after stretch-draw, so as to generate a precompression on a girder tensile area. After prestressing relaxation, the prestressing steel strand has a pressure stress larger than amplitude of a self-weight stress and a fatigue tensile stress; and the girder bottom flange will not be subjected to tension under compound effects of deadweight and fatigue load. Therefore,a fatigue problem id solved. According to the invention, a design fatigue stress amplitude of the bottom flange is increased from a former one limited in an allowed fatigue stress amplitude to one approaching a design strength f, so as o substantially increase anti-fatigue capability and change a fatigue brittle failure to a ductility strength problem. An ultimate strength of the bottom flange isincreased by one time, so as to substantially increase material utilization rate of high-strength steel.