The invention discloses a complex probiotic for glucose regulation and a preparation method. The complex probiotic is composed of autochthonic gordonia powder, synanthrin, microcrystalline cellulose, xylitol, powdered milk, tartary buckwheat powder, oat powder, pumpkin powder, additives and citric acid. The preparation method comprises the steps that A, cultures are cultivated, specifically, the freeze-dried cultures, namely autochthonic gordonia sq-5, are inoculated into an eggplant-shaped bottle bevel surface, and then transferred into an inoculation bottle through sterile water; B, cultivation is conducted in a culture pot, specifically, the cultures in the inoculation bottle are transferred into the culture pot for cultivation; C, cultivation is conducted in a fermentation pot, specifically, culture liquid in the culture pot is injected into the fermentation pot, each thallus is in a single ball arm shape, the bacterial count is 108-1010 individuals/ml, and fermentation is stopped; D, the autochthonic gordonia powder is prepared, specifically, fermentation liquid is boiled, inactivated, concentrated and dried, thalluses are collected, and the autochthonic gordonia powder is obtained; and E, complex probiotic chewable tablets are prepared, specifically, the raw materials are blended and then subjected to wet granulating and tabletting. By adoption of the complex probiotic, toxin and fat can be effectively cleared away, the digestion function is enhanced, the insulin resistance is improved, and glucose and lipid are regulated.