The invention discloses a full-color three-dimensional self-luminous road line paint and a preparation method thereof. The full-color three-dimensional self-luminous road line paint comprises the following components in parts by mass: 20-40 parts of acrylic resin, 0-20 parts of titanium dioxide, 0.1-10 parts of an additive, 10-20 parts of an extender pigment, 10-30 parts of an organic solvent and 20-60 parts of a luminous carrier, wherein the additive is a mixture of a dispersant, a flatting agent, a plasticizer, an anti-settling agent and a defoaming agent at any ratio. The preparation method comprises the following steps: preparing a mother solution by stirring the acrylic resin, the organic solvent and the anti-settling agent at a high speed; respectively adding the titanium dioxide, the extender pigment and the dispersant into the mother liquid and uniformly dispersing by stirring by a medium speed to obtain a predispersoid; grinding the predispersoid, subsequently adding the luminous carrier, the flattening agent, the plasticizer and the defoaming agent and uniformly stirring to obtain the full-color three-dimensional self-luminous road line paint. The method is simple; the prepared paint is safe and nontoxic, has the capabilities of repeatedly absorbing light, storing light and emitting light, and is extremely high in light radiation resistance, high in ageing resistance and good in adhesive force and hardness friction force resistance; a coating formed by the paint is good in water resistance, alkali resistance and weather resistance.