Disclosed herein are a catalyst for hydrocracking reaction of high molecular weight components in bio-oil, a method for preparing the same and a method for bio-oil upgrading using the same. The catalyst includes a zeolite carrier; and at least one metal selected from the group consisting of nickel (Ni), ruthenium (Ru) and cerium (Ce) supported on the carrier. The catalyst promotes the hydrocracking of high molecular weight compounds contained in the bio-oil, but also inhibits the polymerization reaction of the decomposed product, thereby more effectively enhancing the hydrocracking reaction of the bio-oil.