槐定碱抑制内毒素血症小鼠肾组织NF-κB通路活化的研究

Sophoridine inhibits NF-κB signaling pathway activation in kidney tissue of endotoxemia mice

为研究槐定碱干预对内毒素血症小鼠肾组织NF-κB通路的影响,以细菌脂多糖(LPS)引起的内毒素血症肾损伤小鼠为实验对象,槐定碱(12、6及3 mg·kg-1,ip)干预,RT-PCR检测肾组织IKKβ与TNF-α的mRNA表达,免疫组化检测磷酸化IKKβ蛋白(pIKKβ)表达,Western blotting与免疫荧光激光共聚焦显微镜观察肾组织NF-κB的表达与分布,放射免疫法检测血清中TNF-α的水平。结果显示,槐定碱(12、6及3 mg·kg-1)干预降低了内毒素血症小鼠肾组织IKKβmRNA及pIKKβ表达,抑制NF-κB P65蛋白表达并减少NF-κB P65蛋白入核率,从而使肾组织TNF-αmRNA表达及血清TNF-α含量减少。结果提示,槐定碱可抑制LPS引起的炎症反应,其机制可能与抑制NF-κB通路活化有关。

This study is to investigate the effects of sophoridine on NF-κB signaling pathway in kidney tissue of endotoxemia mice and the mechanism involved.BALB/c mice were challenged with lipopolysaccharide（LPS） caudal vein injection,then sophoridine was administered by intraperitoneal injection.Totally 50 mice were divided into 5 groups： control group,LPS model group,sophoridine treatment 12 mg·kg 1 group,6 mg·kg 1 group and 3 mg·kg 1 group.All animals were sacrificed at 6 hours after treatment.Kidney and blood samples were harvested.IKKβ mRNA and TNF-α mRNA expression of renal tissue was measured by the reverse transcription polymerase chain reaction（RT-PCR）,and phosphorylation IKKβ protein（pIKKβ） was detected by immunohistochemistry.NF-κB P65 protein expression and distribution of renal tissue were observed by Western blotting and immunofluorescence laser confocal microscopy.Serum TNF-α level was detected by radioimmunoassay.The results showed that the sophoridine significantly reduced the expression of IKKβ mRNA and pIKKβ protein,and inhibited the expression of NF-κB P65 protein and decreased the entry nuclear rate of NF-κB P65 in the renal tissue of endotoxemia mice.Thereby the renal TNF-α mRNA expression and serum TNF-α level were significantly reduced.These results suggest that sophoridine could inhibit inflammatory reaction induced by LPS through inhibiting activation of NF-κB signaling pathway.