Cell wall polysaccharides (dietary fiber) in cereal grains contribute to health benefits. The novelty of the current study was an effort to explore the in vivo therapeutic potential of different cereal bran cell walls against hypercholesterolemia. For this purpose, the cell walls were isolated from different cereal brans (wheat, maize, oats, and barley), and the intake of these cereal bran cell walls was evaluated for their anti-lipidemic activity in normal and hypercholesterolemic rats. The serum taken from the rats was tested for cholesterol, lipid, and triglyceride profiles before and after treatment. The outcomes of the current study have shown that the cereal cell wall has a significant hypercholesterolemia effect. The biochemical parameters of the control animals were within the normal clinical ranges, indicating that the experimental diets were safe. Among cereal bran cell walls, barley bran significantly decreased cholesterol (56.35 ± 1.35 mg/dL), low-density lipoprotein (56.35 ± 1.05 mg/dL), triglycerides (105.29 ± 1.95 mg/dL), and increased high-density lipoprotein level (48.35 ± 1.35 mg/dL). These findings provide conclusive evidence that the cereal cell wall is beneficial in the treatment of hypercholesterolemia and may potentially provide protection against other acute, recurring, or chronic illnesses.