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目的研究LPS体外刺激滋养层细胞是否诱导表达HBD-2,并进一步探讨该表达与TLR4信号传导通路的关系。方法建立不同孕周的滋养层细胞原代培养体系,应用TLR4阻断剂预处理滋养层细胞30 min前后,给予不同质量浓度的LPS(25、50、100、200、400 ng/ml)作用72 h,采用SYBR GreenⅠ实时荧光定量RT-PCR技术检测滋养层细胞HBD-2 mRNA的表达。结果 1)LPS能诱导滋养层细胞HBD-2 mRNA表达,且这种表达与其具有浓度和时间依赖性;当质量浓度为200 ng/ml,刺激24 h时,HBD-2 mRNA的相对表达量最高;2)TLR4阻断剂能抑制LPS对滋养层细胞表达HBD-2 mRNA的诱导作用,与未阻断之前相比差异具有统计学意义(P<0.01)。结论 LPS可能通过TLR4信号传导通路诱导滋养层细胞表达HBD-2 mRNA,将为宫内感染防治提供新靶点。
Objective To investigate whether LPS stimulation of trophoblast cells in vitro induces the expression of HBD-2 and further explores its relationship with TLR4 signaling pathway. Methods The primary culture system of trophoblast cells was established at different gestation weeks. Twenty-four hours after pretreatment of trophoblast cells with TLR4 antagonist, LPS (25, 50, 100, 200, 400 ng / ml) h, SYBR Green Ⅰ real-time fluorescent quantitative RT-PCR detection of trophoblast HBD-2 mRNA expression. Results 1) LPS induced HBD-2 mRNA expression in trophoblast cells in a concentration-and time-dependent manner. When the concentration of 200 ng / ml LPS stimulated for 24 h, the expression of HBD-2 mRNA was the highest ; 2) TLR4 blockade could inhibit the induction of HBD-2 mRNA expression by trophoblast cells in LPS, which was significantly different from that before non-blockade (P <0.01). Conclusion LPS may induce HBD-2 mRNA expression in trophoblast cells through TLR4 signal transduction pathway, which will provide a new target for the prevention and treatment of intrauterine infection.