Effect Of Lta On Functional Receptor Tlr2Expression In Endometrium Of Mice

In recent years, diseases of high producing dairy cows significantly increased. It has seriously affected development of the dairy industry. As one of the three major diseases of the dairy industry, bovine endometritis incidence rate assumes the trend of escalation. And it seriously impacts on the development of the dairy industry. Bovine endometritis which causes by the gram-positive bacteria is very universal. Lipoteichoic acid is an important component of the cell wall in gram-positive bacteria. Toll-like receptor2, a main receptor of innate immune system, can recognize pathogenic microorganism and play a vital role. It’s also very important to bovine endometritis caused by the gram-positive bacteria. Immune response arose from LTA mediated by TLR2. It recognizes the conserved parts of LTA and activates a signaling cascade that leads to activation the transcription factor, NF-κB, increases the expression of inflammatory cytokines. In this experiment, the expression of TLR2and TNF-α、IL6in mice endometrium which stimulated by LTA in vitro and vivo was analyzed. To provide certain basis for the TLR2signal passage’s research.1Expression of TLR2and TNF-α、IL6mRNA in mice endometrium after LTA stimulation.This experiment took the mice as the experimental objects. Infusion of LTA to cause the endometrium to observes pathological changes. And analyze the changes of the expression of TLR2and TNF-a, IL6. Thirty-six ICR female mice were divided into five test groups and a control group after conception. Mice of five test groups were infused with different concentrations of LTA. And the control group infused with normal saline. Uterine tissues were collected after12hours and the changes were analyzed. The histology results showed that the inflammatory cells increased after different concentrations of LTA stimulation in endometrial tissues. TLR2and TNF-a, IL6mRNA expression were detected after RT-PCR. It indicated LTA can enhance TLR2and LTA TNF-a, IL-6mRNA expression.2Comparison with different methods of endometrial epithelial cells cultureTo explore for mice endometrium epithelial cells culture method,5culture ways were compared:collagenase digestion method, trypsinase and type Ⅰ collagenase digestion method, scraping and type Ⅰ collagenase digestion, tissue culture method and cell mass culture method. The results showed that collagenase digestion method and trypsinase and type Ⅰ collagenase digestion method obtained few cells; Type Ⅰ collagenase digestion and tissue culture method obtained more cells and higher activity, but slower growth; Cell mass culture method obtained higher cell viability, faster growth and higher purity cells. It’s a more appropriate mice endometrium epithelial cells culture method in primary culture.3The expression of TLR2and TNF-α、IL6mRNA in endometrial epithelial cells after LTA stimulation.Establishment of the primary endometrial epithelial cells cultures were divided into test groups and a control group. Test group cells were stimulated with2mL different concentrations of LTA, and2mL DMEM as control. Each group supposed6repeated holes. After3hours stimulation, the medium was removed and the cells were homogenized for RNA extraction. RT-PCR and analysis the expression of TLR2and TNF-a, IL6mRNA were carried out. The experiments were repeated five times. Results showed the mRNA levels of TLR2and TNF-α, IL6changed rapidly after stimulation with LTA, the expression of TLR2and TNFα-, IL6mRNA with0.05μg/mL,0.1μg/mL LTA stimulation were significantly higher than that of control. But the expression with0.5μg/mL LTA stimulation were significantly lower than that of control.