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Abstract Lactobacillus casei Zhang, a potential probiotic strain isolated from homemade koumiss in Inner Mongolia of China, has been sequenced and deposited in GenBank. Real-time quantitative PCR is one of the most widely used methods to study related gene expression levels of Lactobacillus casei Zhang. For accurate and reliable gene expression analysis, normalization of gene expression data using one or more appropriate reference genes is essential. We used three statistical methods (geNorm, NormFinder, and BestKeeper) to evaluate the expression levels of five candidate reference genes (GAPD, gyrB, LDH, 16s rRNA, and recA) under different culture conditions and different growth phases to find a suitable housekeeping gene which can be used as internal standard. The results showed that the best reference gene was GAPD, and a set of two genes, GAPD and gyrB (which were the most stable reference genes), is recommended for normalization of real-time quantitative PCR experiments under all the different experimental conditions tested. The systematic validation of candidate reference genes is important for obtaining reliable analysis results of real-time quantitative PCR studies in gene expression levels of Lactobacillus casei Zhang.

The aim of this study was to evaluate the antidiabetic effects of Lactobacillus casei CCFM0412 on mice with type 2 diabetes induced by a high-fat diet and streptozotocin. Thirty-two male C57 BL/6 J mice were assigned to four groups in this study. Type 2 diabetes was induced by feeding of a high-fat diet and injection of streptozotocin. L. casei CCFM0412 was administered to mice at a dose of 109 cfu/d per mouse for 12 wk. Body weight, fasting and postprandial 2-h blood glucose, oral glucose tolerance, glycosylated hemoglobin, insulin, and glycogen in liver were measured. Endotoxin, tumor necrosis factor-α, and interleukin-10 levels were determined. Lipid metabolic parameters including triglycerides, total cholesterol, high-density lipoprotein cholesterol, and low-density lipoprotein cholesterol were also measured. The activities of glutathione peroxides, reactive oxygen species, and superoxide dismutase, and the levels of glutathione and malondialdehyde in the liver also were determined. Pancreas injury was evaluated by histologic analysis. At 13 wk, L. casei CCFM0412 significantly decreased fasting and postprandial 2-h blood glucose, glycosylated hemoglobin, endotoxin, tumor necrosis factor-α, triglycerides, total cholesterol, low-density lipoprotein cholesterol, reactive oxygen species, and malondialdehyde levels compared with the control group (P < 0.05). The values for insulin, interleukin-10, high-density lipoprotein cholesterol, glutathione peroxides, superoxide dismutase, glutathione, and glycogen were significantly increased at 13 wk (P < 0.05). Islets of Langerhans in the L. casei CCFM0412 group were substantially protected from destruction compared with those in the control group. L. casei CCFM0412 significantly improved glucose intolerance, dyslipidemia, immune-regulatory properties, and oxidative stress in mice with type 2 diabetes induced by a high-fat diet and streptozotocin. The results provide a sound rationale for future clinical trials of oral administration of L. casei CCFM0412 for the primary prevention of type 2 diabetes.

The aim of the present study was to investigate the antiosteoclastogenic effects of black rice (Oryza sativa L.) fermented with Lactobacillus casei (LAB) in RANKL-induced RAW macrophage cells and its antiosteoporosis activity against ovariectomy-induced osteoporosis in rats. LAB extract (LABE) treatment attenuated receptor activator of nuclear factor-kappa B (NF-κB) ligand-induced osteoclastic differentiation in RAW cells by inhibiting intercellular reactive oxygen species generation and downregulating the activation of mitogen-activated protein kinases and NF-κB, leading to the downregulation of c-Fos and expression of nuclear factor of activated T cells c1. This consequently suppressed the expression of osteoclast-specific genes including those for cathepsin K, tartrate-resistant acid phosphatase, calcitonin receptor, and integrin β3. Oral administration of LABE protected against ovariectomy-induced bone loss by significantly inhibiting bone architecture alterations and improving serum bone turnover markers in ovariectomized rats. The findings suggest that the antiosteoporotic activity of LABE may be derived from its antiosteoclastic and anti-bone-resorptive activities. LABE has potential as a promising functional material or substrate to prepare protective agents for osteoporosis and osteoclast-mediated bone diseases.

Peptidoglycan (PG) is a polymer that makes up the cell wall of most bacteria. In this study, the peptidoglycan of Lactobacillus casei ATCC 393 was extracted, and its prebiotic function as well as its effects on intestinal health and inflammation reduction in a colitis murine model were investigated. PG was extracted from L. casei ATCC 393 using the ultrasonic-assisted enzymatic method. A structural characterization and assessment of its antioxidant capacity were subsequently performed to evaluate its functional properties. In a dextran sulfate sodium (DSS)-induced colitis model, dietary supplementation with PG (100 mg/kg) demonstrated significant protective effects. Specifically, the PG intervention group exhibited reduced inflammatory symptoms, improved disease activity indices, suppressed weight loss, and colon shortening compared to the DSS-induced group. Intestinal barrier injury was reversed and the Firmicutes/Bacteroidetes ratio was increased. These clinical improvements were accompanied by decreased circulating levels of pro-inflammatory cytokines (IL-6, TNF-α, and IL-1β). These findings revealed that PG modulated gut microbial ecology by enhancing bacterial diversity and promoting the enrichment of beneficial taxa, particularly the Lachnospiraceae and Lactobacillus species. Additionally, PG intervention increased fecal short-chain fatty acid (SCFA) concentrations, especially the concentration of propionic acid and butyric acid, which increased by 13% and 42%, respectively, compared to the DSS-induced group, suggesting enhanced microbial metabolic activity. Furthermore, these findings emphasize the potential of peptidoglycan as a functional component for preventing colitis through microbial-mediated pathways. This study underscores the prebiotic promise of peptidoglycan in the development of interventions targeting intestinal inflammation and supports its further exploration as a functional agent for promoting human health.