黄芪汤通过TLR4/NF-κB信号通路改善糖尿病肾病大鼠炎症反应的研究

Huangqi decoction improved the inflammatory response of rats with diabetic nephropathy by inhib⁃iting TLR4/NF-κB signal pathway

目的探讨通过建立脂多糖(lipopolysaccharide,LPS)诱导的糖尿病肾病大鼠炎症模型,观察黄芪汤能否通过抑制TLR4/NF-κB信号通路,来改善糖尿病肾病大鼠的炎症因子表达。方法选择48只雄性SD大鼠按随机数字表法分为6组,即对照组、对照组+LPS组、模型组、模型+LPS组、黄芪汤干预组、黄芪汤+LPS组,动物造模采用腹腔注射链脲佐菌素(streptozocin,STZ)。检测大鼠病理、TLR4/NF-κB信号通路相关蛋白及其下游的相关蛋白的表达。结果在糖尿病肾病大鼠和LPS+糖尿病肾病大鼠中的TLR4/NF-κB信号通路明显被激活,TLR4、磷酸化IKKα/β、磷酸化NF-κB等蛋白的表达水平明显增高,其下游的炎症因子的表达也随之升高,肾脏病理指标和纤维化加重。黄芪汤可以改善LPS诱导的糖尿病肾病大鼠炎症模型的病理指标,通过降低TLR4的表达,进而减少IKK和NF-κB的磷酸化,达到抑制TLR4/NF-κB信号通路的作用,使得下游的IL-6、TNF-α、IL-1β等炎症因子的表达水平降低。结论黄芪汤通过下调TLR4/NF-κB信号通路相关蛋白的表达水平改善LPS诱导的糖尿病肾病大鼠炎症模型的炎症反应,改善肾脏病理指标,从而减轻糖尿病肾病大鼠的肾损害,起到延缓糖尿病肾病进展的作用。

Objective To establish an animal model of lipopolysaccharide(LPS)-induced in⁃flammation and observe whether Huangqi Decoction(HD)can improve the inflammatory response in rats with diabetic nephropathy(DN)by inhibiting the TLR4/NF-κB signaling pathway.Methods Fortyeightmale Sprague-Dawley(SD)rats were randomly divided into 6 groups of control,control plus LPS,model,model plus LPS,HD intervention and HD plus LPS.And animal model received an intraperito⁃neal injection of streptozotocin(STZ).Pathology,TLR4/NF-κB signaling pathway related proteins and their downstream related proteins were examined.Results TLR4/NF-κB signaling pathway was signifi⁃cantly activated in DN rats and LPS+DN rats.The expression levels of TLR4,phosphorylated IKKα/β,phosphorylated NF-κB and downstream inflammatory factors were significantly up-regulated.Renal pathological and fibrosis parameters worsened.HD could improve the pathological parameters.It low⁃ered the phosphorylation of IKK and NF-κB by reducing the expression of TLR4 and achievedan inhibi⁃tion of TLR4/NF-κB signal.Thus there was a lower expression of such downstream inflammatory fac⁃tors as IL-6,TNF-αand IL1β.Conclusion HD can improve the inflammatory response by lowering the expression level of TLR4/NF-κB signaling pathway-related protein,improve renal pathological param⁃eters,alleviate renal damage and delay the progression of DN in LPS-induced DN rats.