The invention discloses an airtight adhesive used for tire puncture prevention and a preparation method thereof, belonging to the technical field of tire manufacture and repair. The airtight rubber of the present invention is based on chloroprene rubber, natural rubber, bamboo powder, tackifying resin RX-80, high ammonia natural latex, KN4006 naphthenic rubber oil, rosin, zinc oxide, modified pineapple leaf fiber, accelerator DETU , silane coupling agent Si‑563, dispersant WB212, stearic acid SA, polyphenyl polymethylene polyisocyanate, antioxidant RD, sodium persulfate, biphenyl yellow G as raw materials. The present invention adopts high-ammonia latex, chloroprene rubber with good airtightness and natural rubber with good processability as the mixing system, and the properties of the colloids complement each other. At the same time, the modified pineapple leaf fiber is added to improve the rubber layer and the mouth glue during molding. The viscosity between the materials improves the tensile and tear strength of the rubber, and the various compounding agents cooperate with each other to jointly improve the bonding and repair performance of the product.