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ABSTRACT Gayam seed is one of the native plan of Indonesia, has a high amylose content and is a good source of resistant starch (RS). The autoclaving-cooling cycle treatment is the physical modification to increase RS of starch. Resistant starch has a positive impact on health, such as preventing the increase in blood glucose levels significantly. The high resistant starch in gayam starch is needed for prevention of diabetes mellitus. This study aims to evaluate the antidiabetic properties (blood glucose and SCFA levels) of modified gayamstarchin vivo.The modified gayam starch were prepared by autoclaving-cooling 3 cycles. In vivo studies was prepared as isocaloric feed for three groups of diabetic Sprague Dawley rats induced streptozotocin-nicotinamide (STZ-NA) i.e (a) standar diet AIN 93 M (Control), (b) native gayam starch diet (NGS), and (c) modified gayam starch by autoclaving-cooling for three cycles diet (MGS).evaluation the in vivo effects on blood glucose and SCFA levels.The results showed the autoclaving-coolingcyclestreatment increasedseveral chemical properties i.e. moisture, carbohydrate and amilopectin content, but its decreased fat, starch and amylose content. In vivo studies in 28 days experiments showed that the intervention of modified gayam starch decreased blood glucose and increased SCFA content, were more significant in MGS than NGS diet.It was concluded that that the intervention of modified gayam starch decreased blood glucose and increased SCFA content, were more significant in MGS than NGS diet. Keywords: gayam, modified starch,autoclaving-cooling cycles, antidiabetic, in vivo