Intestinal barrier function, critical for maintaining optimal gut health, is achieved primarily through mucins and tight junction proteins (i.e., zonula occludens-1 and occludin). An aberrant expression of these proteins results in increased paracellular permeability, leading to nonalcoholic steatohepatitis. Furthermore, enhanced expression of the inflammasome’s nucleotide-binding domain, leucine-rich repeat-containing protein 3 (cryopyrin), and Toll-like receptor 4 are also associated with nonalcoholic steatohepatitis. To understand the role of probiotic supplementation (Lactobacillus acidophilus, Bifidobacterium longum, and Streptococcus thermophilus) in rats on a high-fat, high-fructose diet with nonalcoholic steatohepatitis, the expression of occludin, cryopyrin, and Toll-like receptor 4 was evaluated. A comparison of the results between the control, nonalcoholic steatohepatitis group, and probiotic-treated nonalcoholic steatohepatitis groups showed (a) a lack of any significant difference in occluding expression (P = 0.724) and (b) a significant reduction in serum Toll-like receptor 4 (P = 0.012) and cryopyrin (P = 0.025) by probiotics. Furthermore, only one rat developed nonalcoholic steatohepatitis in the probiotic group, compared to six rats in the non-probiotic group. In conclusion, there were hepatoprotective effects of probiotic administration on high-fat, high-fructose diet-induced nonalcoholic steatohepatitis in Sprague-Dawley rats.