Accumulation of triglyceride (TG) in enterocytes of the small intestine follows two specific absorption pathways: the monoacylglycerol acyltransferase pathway (MGAT) and glycerol acyltransferase pathway (GPAT), the latter (GPAT) is usually found in the small intestine. In this study, we investigated the effects of Lactobacillus rhamnosus MRS6AN, an isolate from “amabere amaruranu” a Kenyan traditional cultured milk, on triglyceride accumulation and expression of GPAT3, I-FABP, MTP and NPC1L1 in Caco-2 cell enterocyte model. Intracellular triglyceride level (TG) of Caco2 cells was significantly reduced by live L. rhamnosus (LB) compared with other bacterial products. MTP expression in Caco2 cells was minimally reduced in live L. rhamnosus (LB) treated Caco2 cells. However, the expression level GPAT3, I-FABP and NPC1L1 was reduced in Caco2 cells treated with live bacteria. Data from this study suggests that Lactobacillus rhamnosus MRS6AN may reduce lipid uptake and accumulation perhaps via modulation of GPAT3 pathway.