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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 investigate the effect of E. coli lipopolysaccharide (LPS) on the activation of NF-κB in rat Kupffer cells. Methods Kupffer cells were randomly divided into A, B, C three groups. Group A was control group. Group B received LPS supplemented with Kupffer cell culture medium and group C co-cultured PDTC with LPS plus Kupffer cell culture medium. The activity of NF-κB was detected by EMSA, the content of I-κB was detected by Western blot, and the expression of TNF-α mRNA and IL-6 mRNA was detected by RT-PCR. Results Kupffer cells increased the activity of NF-κB, the level of I-κB decreased and the expression of TNF-αmRNA and IL-6mRNA increased in LPS-treated Kupffer cells. The Kupffer cells decreased the activity of NF- κB levels increased, TNF-αmRNA, IL -6mRNA expression decreased. Conclusion LPS can induce the activation of NF-κB, and promote the expression of KCs-derived neurotrophic factor to promote cell injury.