北京中医药大学学报2025，Vol.48Issue(1)：21-33,13.DOI:10.3969/j.issn.1006-2157.2025.01.004
益肾通络方调控TLR4/MyD88/NF-κB信号通路改善糖尿病肾病小鼠细胞焦亡的机制研究
Mechanism of Yishen Tongluo Formula regulating the TLR4/MyD88/NF-κB signaling pathway to ameliorate pyroptosis in diabetic nephropathy mice
摘要
Abstract
Objective To investigate the mechanism of Yishen Tongluo Formula in ameliorating renal pyroptosis in diabetic nephropathy mice by regulating the toll-like receptor 4(TLR4)/myeloid differentiation factor 88(MyD88)/nuclear factor-κB(NF-κB)signaling pathway.Methods Sixty C57BL/6 male mice were randomly divided into control(10 mice)and intervention groups(50 mice)using random number table method.The diabetes nephropathy model was established by intraperitoneally injecting streptozotocin(50 mg/kg).After modeling,the intervention group was further divided into model,semaglutide(40 μg/kg),and high-,medium-,and low-dose Yishen Tongluo Formula groups(15.6,7.8,and 3.9 g/kg,respectively)using random number table method.The high-,medium-,and low-dose Yishen Tongluo Formula groups were administered corresponding doses of medication by gavage,the semaglutide group received a subcutaneous injection of semaglutide injection,and the control group and model groups were administered distilled water by gavage for 12 consecutive weeks.Random blood glucose levels of mice in each group were monitored,and the 24-h urinary protein content was measured using biochemical method every 4 weeks;after treatment,the serum creatinine and urea nitrogen levels were measured using biochemical method.The weight of the kidneys was measured,and the renal index was calculated.Hematoxylin and eosin,periodic acid-Schiff,periodic Schiff-methenamine,and Masson staining were used to observe the pathological changes in renal tissue.An enzyme-linked immunosorbent assay was used to detect urinary β2-microglobulin(β2-MG),neutrophil gelatinase-associated lipocalin(NGAL),and kidney injury molecule-1(KIM-1)levels.Western blotting and real-time fluorescence PCR were used to detect the relative protein and mRNA expression levels of nucleotide-binding domain leucine-rich repeat and pyrin domain-containing receptor 3(NLRP3),Caspase-1,gasdermin D(GSDMD),interleukin-1β(IL-1β),and interleukin-18(IL-18)in renal tissue.Immunohistochemistry was used to detect the proportion of protein staining area of the TLR4,MyD88,and NF-κB in renal tissue.Results Compared with the control group,the random blood glucose,24-h urinary protein,serum creatinine,urea nitrogen,and renal index of the model group increased,and the urine β2-MG,NGAL,and KIM-1 levels increased.The relative protein and mRNA expression levels of NLRP3,Caspase-1,GSDMD,IL-1β,and IL-18 in renal tissue increased,and the proportion of TLR4,MyD88,and NF-κB protein positive staining areas increased(P<0.05).Pathological changes such as glomerular hypertrophy were observed in the renal tissue of the model group.Compared with the model group,the Yishen Tongluo Formula high-dose group showed a decrease in random blood glucose after 12 weeks of treatment(P<0.05).The Yishen Tongluo Formula high-and medium-dose groups showed a decrease in 24-h urinary protein,creatinine,urea nitrogen,and renal index,as well as decreased β2-MG,NGAL,and KIM-1 levels.NLRP3,Caspase-1,GSDMD,IL-1 β,and IL-18 relative protein and mRNA expression levels were also reduced,and the proportion of TLR4,MyD88,and NF-κB protein positive staining areas was reduced(P<0.05).Pathological damage to renal tissue was ameliorated.Conclusion Yishen Tongluo Formula may exert protective renal effects by inhibiting renal pyroptosis and alleviating tubular interstitial injury in diabetic nephropathy mice by regulating the TLR4/MyD88/NF-κB signaling pathway.关键词
糖尿病肾病/益肾通络方/细胞焦亡/Toll样受体4/髓样分化因子88/核转录因子-κB信号通路/小鼠Key words
diabetes nephropathy/Yishen Tongluo Formula/pyroptosis/toll-like receptor 4/myeloid differentiation primary response protein 88/nuclear factor-κB signaling pathway/mice分类
医药卫生引用本文复制引用
张轶斐,郭玺,刘忠杰,刘伟敬,曹梓静,张泽钰,白雪慧,唐靖怡,席俊羽,王嘉奕,谢亦冉,吴宇琪..益肾通络方调控TLR4/MyD88/NF-κB信号通路改善糖尿病肾病小鼠细胞焦亡的机制研究[J].北京中医药大学学报,2025,48(1):21-33,13.基金项目
国家自然科学基金面上项目(No.82374382,No.82074361,No.82274293) （No.82374382,No.82074361,No.82274293）
中央高校基本科研业务费专项资金资助项目揭榜挂帅项目(No.2023-JYB-JBZD-037) （No.2023-JYB-JBZD-037）
国家中医药管理局高水平医院临床研究和成果转化能力提升试点项目揭榜挂帅项目(No.DZMG-XZYY-23002) （No.DZMG-XZYY-23002）
创新中药关键技术国家重点实验室开放课题(No.ZSL-003-02) National Natural Science Foundation of China(Nos.82374382,82074361,and 82274293) （No.ZSL-003-02）
