The invention discloses an anti-aging and anti-fatigue traditional Chinese medicinal formula and application thereof. The traditional Chinese medicinal formula comprises the following components in parts by weight: 5-20 parts of lucid ganoderma, 5-15 parts of panax notoginseng, 10-30 parts of radix acanthopanacis senticosi, 5-25 parts of root of Kirilow Rhodiola, 10-25 parts of fructus lycii, 5-15parts of radix astragali, 15-35 parts of fructus Ziziphi Jujubae, 5-20 parts of rhizoma polygonati, 15-30 parts of cornu cervi pantotrichum, 5-15 parts of fructus schizandrae, 5-15 parts of rhizoma chuanxiong, 10-25 parts of rhizoma gastrodiae, 5-15 parts of poria cocos, 5-20 parts of flos chrysanthemi, 10-30 parts of fiveleaf gynostemma herb and 5-12 parts of cyclocarya paliurus. The traditionalChinese medicinal formula is applied to preparation of anti-aging and anti-fatigue medicines. Through a synergistic effect of various medicinal materials in a traditional Chinese medicine prepared according to the traditional Chinese medicinal formula provided by the invention, the traditional Chinese medicine has the effects of resisting fatigue, resisting aging, improving immunity, treating insomnia, clearing liver heat, improving eyesight, resisting bacteria, eliminating inflammation and resisting cancers, and is mild in medicinal nature, significant in effect and free of side effects; inaddition, the conventional auxiliary materials are added into the traditional Chinese medicinal formula according to the conventional technology, and clinically accepted capsules, tablets, pills, granules or oral liquid can be prepared, so that the using universality is improved.