The invention relates to injection amphiphilic-microsphere-containing hyaluronic acid mixed gel and a preparation method thereof. The microsphere material relates to amphiphilic materials of PLLA-PEG, PLGA, PLGA-PEG and the like. The molecular weight of PLLA or PLGA is 10,000-500,000, the molecular weight of PEG is 1,000-10,000, and in PLGA, LA/GA=90/10-10/90. The average grain diameter of the microsphere is 1-200 [mu[m, and the mass percent of the microsphere in the gel is 1-50%; the gel is made from hyaluronic acid, divinyl sulphone crosslinked hyaluronic acid, or glycidyl ether crosslinked hyaluronic acid, the molecular weight of the hyaluronic acid is 100,000-3,000,000, and the mass percent of the hyaluronic acid is 1-50%. The gel can be also made from animal-derived collagen, chitosan, amino acid cellulose and sodium alginate. The microsphere mixed gel is prepared in a sodium chloride solution or a phosphate buffered solution with the osmotic pressure of 250-350 mOsm/L and pH of 6.5-7.5. According to the invention, the amphiphilic microsphere is directly mixed with hyaluronic acid gel, can be uniformly distributed in the gel easily, and is unlikely to aggregate into blocks in the water environment of a human body after being injected.