大肠杆菌脂多糖诱导大鼠Kupffer细胞NF-κB激活及其意义

Significance of NF-κB Activation Induced by LPS in Rat Kupffer Cells

目的 探讨大肠杆菌脂多糖 (LPS)诱导大鼠Kupffer细胞NF -κB激活及其意义。方法 将Kupffer细胞随机分为A、B、C三组。A组为对照组 ,B组将LPS加入Kupffer细胞培养基共同培养 ,C组将PDTC和LPS加Kupffer细胞培养基共同培养。用EMSA法检测NF -κB活性 ,用Westernblot法检测I-κB蛋白含量 ,用RT -PCR法检测TNF -αmRNA、IL -6mRNA表达。结果 培养液中加入LPS后Kupffer细胞NF -κB活性增加 ,I -κB水平下降 ,TNF -αmRNA、IL -6mRNA表达增加 ;含有LPS的培养液中加入PDTC后Kupffer细胞NF -κB活性减少 ,I-κB水平上升 ,TNF -αmRNA、IL -6mRNA表达减少。结论 LPS可诱导NF -κB激活 ,并促进KCs源性细胞损害递质的表达从而促进细胞损伤。

Objective To explore the effect of NF-κB activation induced by lipopolysaccharide(LPS) in rat Kupffer cells(KCs). Methods KCs were divided randomly into three groups. A group was control group. KCs were co-cultured with LPS in B group. KCs were co-cultured with LPS and PDTC in C group. NF-κB activity of KCs was determined with EMSA. Content of I-κB protein in KCs was detected with western blot. Expression levels of TNF-α mRNA, IL-6 mRNA of KCs were measured with RT-PCR. Results LPS induced NF-κB activation of KCs, decreased I-κB content of KCs, and increased expression levels of TNF-α mRNA and IL-6 mRNA markedly. PDTC could diminish the above effects of LPS on KCs. Conclusions NF-κB activation induced by LPS may induce cell injury by promoting expressions of TNF-α mRNA and IL-6 mRNA in KCs. PDTC could decrease mRNA expression of proinflammatory mediators by suppressing excessive NF-κB activation in KCs.