Parallel with the recent increasing interest in alternative/herbal medicine for the prevention and treatment of various diseases including hypercholesterolemia, in addition to the increasing uses of Gum Arabic (GA) as one option. The objective of this study was to evaluate the effect of Gum Arabic mixed diet on serum concentration of total cholesterol (TC), very low density lipoprotein cholesterol (VLDL-c), low density lipoprotein cholesterol (LDL-c), high density lipoprotein cholesterol (HDL-c), and triacylglycerol (TAG) in rats fed diet supplemented with cholesterol. Twenty five Wistar albino rats were divided into five groups named A, B, C, D, and E, each of five rats. Group A was fed on basal diet and served as control, group B received the basal diet mixed with 1% cholesterol powder. Gum Arabic (GA) was added to the basal diet with 1% cholesterol powder at the rate of 10%, 20% and 30% in groups C, D and E, respectively. After 30 days, blood samples were collected from all groups for analysis. There was non-significant reduction on TC in groups C, D and E compared to group B. The level of LDL-c was found to be significantly (P<0.05) decreased in groups C, D and E compared to group B. In groups C, D and E HDL-c level was significantly (P<0.05) increased compared to group B. Serum TAG and VLDL-c were reduced in dose response manner of Gum Arabic in groups C, D and E compared to group B, although the decrease was only significant (P< 0.05) in groups D and E. The present study concluded that the addition of Gum Arabic to rat's diet has positive effect in lowering serum cholesterol and triacylglycerol levels.