乳酸杆菌脂磷壁酸抑制脂多糖诱导的大鼠肝脏Kupffer细胞TLR4通路的激活

Lipoteichoic Acid of Lactobacillus Bulgaricus Inhibits the Lipopolysaccharide-activated TLR4 Pathway in Rat Kupffer Cells

目的：探讨保加利亚乳酸杆菌脂磷壁酸（Lipoteichoic Acid of lactobacillus bulgaricus, LBG-LTA）对大鼠肝脏Kupffer细胞Toll样受体4（Toll-like receptor4，TLR4）信号通路的作用。方法：雄性健康Wistar大鼠10只（2月龄，体重250～300g）处死后。分离培养肝脏Kupffer细胞；培养LBG，并提取制备LBG-LTA；Kupffer细胞，在有或无LBG-L11A（0．1、1、10wg／mL）预处理的情况下，给予脂多糖（lipopolysaccharide，LPS，1EU／mL）刺激后，Westernblot检测各孔Kupffer细胞的TLR4、TANK结合激酶1（TANK binding kinase-1，TBK1）及核中的核因子B（nuclear factor-κB，NF-κB）水平，酶联免疫吸附法检测各孔培养上清中的肿瘤坏死因子α（tumornecrosisfactor-α，TNF-α）和白介素1β（interleukin-1β，IL-1β）。结果：分离的Kupffer细胞经不同浓度LBG-LTA预处理后，其在LPS刺激下所表达的TLR4、TBK1、核中NF-B的水平及生成的TNF-a和IL-1β明显低于无LBG-LTA预处理情况下的LPS孔（P〈0．05），且LBG-LTA的作用呈浓度依赖性。结论：LBG-LTA以浓度依赖的方式抑制了LPS诱导下大鼠Kupffer细胞TUM通路的激活。

Objecfive： To determine whether Lipoteichoic Acid of lactobacillus bulgaricus （LBG-LTA） inhibits ToU-like receptor 4 （TLR4） pathway in Kupffer cells isolated from rats. Methods： Ten healthy male Wistar rats, 2 months old, 250-300 g, were killed for the isolation of Kupffer ceils. Isolated Kupffer cells were treated with lipopolysaccharide （LPS, 1 EU/mL） with or without LBG-LTA （0.1, 1 or 10μg/mL） pretreatment. The expression of TLR4, TANK binding kinase-1 （TBK 1） and activated nuclear factor-κB （NF- B） in Kupffer cells was measured by Western blot. The production of tumor necrosis factor-α （TNF-α） and interleukin-1β （IL-1β） was quanti- fied by ELISA. Results： The LPS-induced expression of TLR4, TBK1 and activated NF- B and production of TNF-α and IL-1β in isolat- ed Kupffer cells were significantly inhibited by LBG-LTA pretreatment in a dose-dependent manner （P〈0.05）. Conclusions： LBG-LTA inhibits LPS-activated TLR4 pathway in Kupffer ceils of rats in a dose-dependent manner.