基于TLR4/NF-κB p65/NLRP3信号通路探讨当归芍药散改善糖尿病肾病小鼠炎症反应机制OA
Mechanism of Danggui Shaoyaosan in Improving Inflammatory Response in Mice with Diabetic Kidney Disease Based on TLR4/p65/NLRP3 Signaling Pathway
目的:观察当归芍药散对自发性糖尿病db/db小鼠肾脏组织中Toll样受体4/核转录因子-κB p65/NOD样蛋白受体3(TLR4/NF-κB p65/NLRP3)信号通路的表达,探讨当归芍药散缓解糖尿病肾病(DKD)炎症的可能作用机制.方法:将30只db/db小鼠按随机数字表法分为5组(n=6):模型组,当归芍药散低(16.77 g·kg-1·d-1)、中(33.54 g·kg-1·d-1)、高(67.08 g·kg-1·d-1)3个剂量干预组及厄贝沙坦组(0.025 g·kg-1·d-1);同时,以db/m小鼠(n=6)作为正常组.干预8周后,检测小鼠体质量、空腹血糖(FBG)、24h尿蛋白(24 h-UTP)及血肌酐浓度(SCr);苏木素-伊红(HE)染色、马松(Masson)染色和过碘酸-希夫染色(PAS)染色对肾脏组织进行病理学观察和分析;蛋白免疫印迹法(Western blot)、实时荧光定量聚合酶链式反应(Real-time PCR)检测TLR4、NF-κB p65、NLRP3和肿瘤坏死因子-α(TNF-α)、白细胞介素(IL)-1β、IL-6、IL-10、IL-18蛋白及mRNA表达水平;免疫组化法(IHC)检测肾脏组织中TLR4、NF-κB p65、NLRP3蛋白的表达.结果:与正常组比较,模型组小鼠体质量、FBG、24 h-UTP、SCr表达升高(P＜0.05);肾脏结构紊乱,基底膜增厚,间质可见较多炎细胞浸润;TLR4、NF-κB p65、NLRP3、TNF-α、IL-1β、IL-6、IL-18表达增加,IL-10表达降低(P＜0.05);与模型组比较,给药组大鼠FBG、24 h-UTP、SCr水平均有不同程度降低,肾脏组织病理损伤明显改善,TLR4、NF-κB p65、NLRP3、TNF-α、IL-1β、IL-6、IL-18表达降低,IL-10表达升高(P＜0.05).结论:当归芍药散可能通过调节TLR4/NF-κB p65/NLRP3信号通路减轻肾脏炎症反应并降低尿蛋白排泄,进而延缓DKD的疾病进展.
Objective:To investigate the effect of Danggui Shaoyaosan on the expression of Toll-like receptor 4/nuclear factor-kappa B p65/NOD-like receptor protein 3(TLR4/NF-κB p65/NLRP3)signaling pathway in the renal tissues of db/db mice with spontaneous diabetes,and to explore the potential mechanism by which Danggui Shaoyaosan alleviates inflammation in diabetic kidney disease(DKD).Methods:Thirty db/db mice were divided into five groups:A model group,Danggui Shaoyaosan low-(16.77 g·kg-1·d-1),medium-(33.54 g·kg-1·d-1),and high-dose(67.08 g·kg-1·d-1)intervention groups,as well as an irbesartan group(0.025 g·kg-1·d-1)by the random number table method,with 6 mice in each group.Additionally,6 db/m mice were assigned to the normal group.After 8 weeks of intervention,the following parameters were determined by corresponding methods:body weight,fasting blood glucose(FBG),24-hour urinary protein(24 h-UTP),and serum creatinine(SCr)levels,renal histopathological analysis by hematoxylin-eosin(HE)staining,Masson staining,and periodic acid-Schiff(PAS)staining,the protein and mRNA expression levels of TLR4,NF-κB p65,NLRP3,tumor necrosis factor-alpha(TNF-α),interleukin-1β(IL-1β),interleukin-6(IL-6),interleukin-10(IL-10),and interleukin-18(IL-18)by Western blot and Real-time quantitative polymerase chain reaction(Real-time PCR),as well as TLR4,NF-κB p65,and NLRP3 protein expression in renal tissues by immunohistochemistry(IHC).Results:Compared with the normal group,the model group exhibited increased body weight,FBG,24 h-UTP,and SCr levels(P＜0.05);disordered renal structure,thickened basement membrane,and interstitial inflammatory cell infiltration,elevated TLR4,NF-κB p65,NLRP3,TNF-α,IL-1β,IL-6,and IL-18 expression;as well as decreased IL-10 expression(P＜0.05).Compared with the model group,these pathological changes and biochemical abnormalities were reversed in the medicine intervention groups to varying degrees(P＜0.05).Conclusion:Danggui Shaoyaosan may delay DKD progression by alleviating renal inflammatory response and reducing urinary protein excretion via modulating the TLR4/NF-κB p65/NLRP3 signaling pathway.
郭世龙;李睿嘉;王子轩;王新爱;侯露瑜;石文婧;田萌媛;郭登洲
河北中医药大学研究生学院,石家庄 050091河北中医药大学研究生学院,石家庄 050091河北中医药大学研究生学院,石家庄 050091河北中医药大学第一附属医院/河北省中医院,石家庄 050011河北中医药大学研究生学院,石家庄 050091河北中医药大学研究生学院,石家庄 050091河北中医药大学第一附属医院/河北省中医院,石家庄 050011河北中医药大学第一附属医院/河北省中医院,石家庄 050011
医药卫生
糖尿病肾病当归芍药散炎症反应Toll受体4/核转录因子-κB二聚体基因/NOD样蛋白受体3通路db/db小鼠
diabetic kidney diseaseDanggui Shaoyaosaninflammatory responseToll-like receptor 4/nuclear factor-kappa B p65/NOD-like receptor protein 3(TLR4/NF-κB p65/NLRP3)signaling pathwaydb/db mice
《中国实验方剂学杂志》 2026 (6)
19-27,9
河北省中医药管理局科研计划项目(2025285)河北省自然科学基金项目(H2022423367)河北中医药大学研究生创新项目(XCXZZSS2025024)
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