参芪地黄汤加味对脂多糖诱导人肾小管上皮细胞TLRs/NF-κB信号通路的影响  

Effect of Shenqi Dihuang decoction on lipopolysaccharide induced TLRs/NF-κB signaling pathway in HK-2 cells

作　　者：吴斯琦 丁萌譞 舒占钧[2] WU Siqi;DING Mengxuan;SHU Zhanjun(The Forth Clinical College,Xinjiang Medical University,Urumqi 830099,China;The Fourth Affiliated Hospital of Xinjiang Medical University,Urumqi 830099,China)

机构地区：[1]新疆医科大学第四临床医学院,新疆乌鲁木齐830099 [2]新疆医科大学第四附属医院,新疆乌鲁木齐830099

出　　处：《陕西中医》2025年第3期320-324,共5页Shaanxi Journal of Traditional Chinese Medicine

基　　金：新疆维吾尔自治区重点实验室开放课题(2023D04075)。

摘　　要：目的:通过体外细胞实验研究参芪地黄汤加味对脂多糖(LPS)诱导的人肾小管上皮细胞(HK-2细胞)Toll样受体/核因子-κB(TLRs/NF-κB)信号通路的影响。方法:取雄性SD大鼠20只,随机分为对照组、给药组。对照组给予等体积0.9%氯化钠溶液灌胃;给药组给予参芪地黄汤加味灌胃,连续给药7 d。干预后采集血液,用HK-2细胞专用培养基培养HK-2细胞。将细胞分为对照组(正常大鼠血清)、脂多糖组(正常大鼠血清与脂多糖)、参芪地黄汤低浓度组(脂多糖与5%参芪地黄汤加味含药血清)、参芪地黄汤高浓度组(脂多糖与10%参芪地黄汤加味含药血清)。各组培养24、48 h后收获细胞。酶联免疫法(ELISA)检测白介素6(IL-6)、肿瘤坏死因子α(TNF-α);蛋白质印迹法(Western blotting)检测Toll样受体2(TLR2)、Toll样受体4(TLR4)、髓样分化因子88(MyD88)、核因子-κB(NF-κB)表达。结果:与对照组相比,脂多糖组IL-6、TNF-α水平升高,经参芪地黄汤加味干预后IL-6、TNF-α降低,差异有统计学意义(均P<0.05)。与对照组、脂多糖组相比,TLR4、MyD88、NF-κB p65降低,差异有统计学意义(均P<0.05)。结论:参芪地黄汤加味含药血清可下调炎症水平,抑制TLRs/NF-κB信号通路,改善脂多糖诱导炎症,改善CKD微炎症,保护HK-2细胞,延缓CKD进程。Objective:To investigate effect of Shenqi Dihuang decoction on lipopolysaccharide(LPS)-induced Toll-like receptor/nuclear factor-κB(TLRs/NF-κB)signaling pathway in human renal tubular epithelial cells(HK-2 cells)through in vitro cell experiments.Methods:20 male SD rats were randomly divided into control group and administration group.The control group were gavaged with the same volume of 0.9%sodium chloride solution.The administration group were gavaged with Shenqi Dihuang decoction 7 consecutive days.Blood were collected after the intervention.HK-2 cells were cultured with medium dedicated to HK-2 cells.The cells were divided into control group(normal rat serum),lipopolysaccharide group(normal rat serum+lipopolysaccharide),Shenqi Dihuang low-concentration group(lipopolysaccharide+5%Shenqi Dihuang decoction),and Shenqi Dihuang high-concentration group(lipopolysaccharide+10%Shenqi Dihuang decoction).Cells were harvested after 24 h and 48 h of culture.The levels of interleukin 6(IL-6)and tumor necrosis factorα(TNF-α)were detected by enzyme-linked immunoassay(ELISA).Expression levels of Toll-like receptor 2(TLR2),Toll-like receptor 4(TLR4),myeloid differentiation factor 88(MyD88)and nuclear factor-κB(NF-κB)were detected by western blotting.Results:Compared with the control group,levels of IL-6 and TNF-αin the lipopolysaccharide group were significantly increased,and levels of IL-6 and TNF-αdecreased after intervention of Shenqi Dihuang decoction,difference statistically significant(all P<0.05).The expression levels of TLR4,MyD88 and NF-κB p65 were significantly decreased compared with the control group and lipopolysaccharide group,difference statistically significant(all P<0.05).Conclusion:The medicated serum of Shenqi Dihuang decoction can down-regulate level of inflammation,improve inflammatory damage induced by lipopolysaccharide,improve micro-inflammatory state of CKD,protect HK-2 cells,and delay process of CKD by inhibiting the expression of TLRs/NF-κB signaling pathway.

关 键 词：慢性肾脏病 参芪地黄汤 脂多糖 TOLL样受体 核因子-κB通路 微炎症状态 

分 类 号：R256.5[医药卫生—中医内科学]