# Physiological Effects of Hydrolyzed Skim Milk and Probiotics on Osteoporosis Models
## 3 Results and Discussion
### Effect of Skin Milk and Probiotics on the Small Intestine
reported that Muc2-/-mice had higher susceptibility to attaching and effacing one of the mucosal pathogens, _Citrobacter rodentium_[36]; these mice also showed clinical signs of colitis. Muc2-/-mice exhibited mucosal thickening, superficial erosion, and hyperproliferation [37]. Prior studies strongly suggest that mucin 2 is critical for gut health. Furthermore, our results clearly show that the intake of skim milk and skim milk with probiotics can improve intestinal health by increasing mucin 2 production in the intestine. In particular, the expression of the Mucin2 gene was more pronounced in the co-administration group, which involved the administration of skim milk and probiotics, compared with that in the skim-milk-only group. A previous study revealed that human-milk-oligosaccharide-treated cells exhibit increased mucin 2 expression, decreased bacterial attachment, and increased dextran permeability when challenged with enteric pathogens [25]. In addition, several types of _Lactobacillus_ reportedly increase the expression of mucin protein in HT29 cells, a human intestinal cell line [38]. An extract of _L. acidophilus_ A 4 cell increased mucin 2 expression  
Figure 4: Expression of occludin and mucin 2 in the small intestines of the control, ovariectomy (OVX), and skim-milk-and-probiotic groups. The mRNA expression levels of occludin (**A**) and mucin 2 (**B**) in the intestine of each experimental group. Immunohistochemistryidochemical analysis of the mucin 2 protei. Scale bar = 50 μm (**C**). Western blotting analysis of the occludin protein; the graph shows the results of densitometric analysis of the band normalized to that of \(\beta\)-actin (**D**). The results are presented as mean \(\pm\) SD. * \(p<0.05\) and ** \(p<0.01\)** were considered to indicate statistical significance. Data from the experimental group were compared with data from the OVX group. in HT29 cells [39]. These studies highlight the positive synergistic effect of simultaneous intake on the increase in Mucin 2 in our study. The intestine is a very important organ that integrates immunity, nutrient absorption, and digestion. TJs, which form in the intestinal epithelia, play a critical role in the physical intestinal barrier and modulate the transfer of various substances. Among TJs, occludin is abundantly expressed at cell-to-cell contact sites. Occludin-/-mice exhibited increased inflammation, growth retardation, and hyperplasia, despite normal TJ structure in the intestine [40]. Accordingly, increased occludin levels improve the TJ barrier function and prevent TJ damage [41; 42]. Based on our results, the protein levels of occludin were markedly increased in the skim-milk and skim-milk-and-probiotic groups compared to those in the OVX groups, indicating that the intake of skim milk and probiotics has positive effects on intestinal TJ function. In addition, comparative analysis of the various effectiveness mentioned above with fermented skim milk using probiotics will be necessary as further study.