中医药信息2025，Vol.42Issue(4)：1-11,11.DOI:10.19656/j.cnki.1002-2406.20250401
通络保肾方调控NLRP3/GSDMD通路抑制细胞焦亡改善IgA肾病大鼠足细胞损伤的机制
Mechanistic Study of Tongluo Baoshen Formula in Ameliorating Podocyte Injury in IgA Nephropathy Rats by Regulating NLRP3/GSDMD Pathway to Inhibit Pyroptosis
摘要
Abstract
Objective:To observe the effect of Tongluo Baoshen Formula(TLBSF)on improving podocyte injury in IgA nephropathy(IgAN)rats and explore its mechanism.Methods:A total of 130 SD rats were randomly divided,with 20 rats in the normal group and 110 in the modeling group.The IgAN model was established using bovine serum albumin,carbon tetrachloride,and lipopolysaccharide.Finally,100 model rats were successfully constructed and randomly divided into five groups:model group,losartan potassium group[5×10-3 g/(kg·d)],and low,medium,and high-dose TLBSF groups[5.3,10.6,21.2 g/(kg·d)],each containing 20 rats.The treatment groups were administered the respective doses via gavage,while the normal and model groups received equal volumes of distilled water.After 4 and 8 weeks of treatment,samples were collected from 10 randomly selected rats per group at each time point.ELISA and creatinine oxidase methods were used to detect the urinary albumin-to-creatinine ratio(ACR),interleukin-18(IL-18),caspase-1,gasdermin D(GSDMD),and blood IgA levels.NOD-like receptor protein 3(NLRP3),GSDMD-N-terminal(GSDMD-NT),IL-18,integrin α3,and integrin β1 mRNA and protein expression levels in kidney tissue were measured using real-time quantitative PCR and Western blot.Immunohistochemistry was used to detect NLRP3 and caspase-1 expression in kidney tissue,and immunofluorescence was employed to observe GSDMD-NT expression.Transmission electron microscopy was utilized to assess podocyte ultrastructural changes.Results:Compared with the normal group,the model group showed significantly increased urinary ACR,caspase-1,GSDMD,and IL-18 levels(P<0.01),which progressively increased with prolonged modeling duration(P<0.05 or P<0.01).However,blood IgA levels did not differ significantly(P>0.05).NLRP3,GSDMD-NT,IL-18 mRNA,and protein expression levels were significantly elevated in kidney tissue(P<0.01),while integrin α3 and integrin β1 mRNA and protein levels were significantly reduced(P<0.01),with levels further decreasing over time(P<0.01).Immunohistochemistry showed a significantly increased NLRP3 and caspase-1 positive area(P<0.01),primarily localized in glomerular capillary loops and mesangial regions,with limited expression in renal tubules.Immunofluorescence showed significantly enhanced GSDMD-NT fluorescence intensity(P<0.01),primarily expressed in glomerular capillary loops and mesangial regions.Ultrastructurally,podocytes exhibited enlargement,partial foot process fusion,microfilament reduction,lysosomal membranous structures,and segmental membrane rupture.Compared with the model group,the low,medium,and high-dose TLBSF groups and the losartan potassium group showed significantly reduced urinary ACR,IL-18,and caspase-1 levels(P<0.01)at 4 and 8 weeks of treatment.NLRP3,GSDMD-NT,IL-18 mRNA,and protein levels were significantly reduced(P<0.01),while integrin α3 and integrin β1 levels were significantly increased(P<0.01).Immunohistochemistry revealed reduced NLRP3 and caspase-1 positive areas(P<0.05 or P<0.01),and immunofluorescence showed decreased GSDMD-NT fluorescence intensity(P<0.05 or P<0.01).Ultrastructurally,podocyte repair was observed,with reduced foot process fusion and alleviated swelling.High-dose TLBSF showed the most significant improvement,with further enhancement after 8 weeks compared to 4 weeks.Conclusion:TLBSF improves podocyte injury in IgAN rats,potentially through regulating the NLRP3/GSDMD pathway to inhibit pyroptosis.关键词
通络保肾方/IgA肾病/足细胞损伤/NLRP3/GSDMD通路/细胞焦亡Key words
Tongluo Baoshen Formula/IgA nephropathy/Podocyte injury/NLRP3/GSDMD pathway/Pyroptosis引用本文复制引用
刘永芳,周莉,刘晖扬,代建风,刘英华,陈帮明,林雪菲,杨太旺,刘星宇..通络保肾方调控NLRP3/GSDMD通路抑制细胞焦亡改善IgA肾病大鼠足细胞损伤的机制[J].中医药信息,2025,42(4):1-11,11.基金项目
国家自然科学基金项目(82060839) （82060839）
江西省中医肾病临床医学研究中心建设项目(2019BCD42006) （2019BCD42006）
国家中医优势专科(肾病科)建设项目(赣中医药综合字[2023]3号) （肾病科）
江西省中医优势病种肾衰病防治中心建设项目(赣中医药医政字[2021]8号) （赣中医药医政字[2021]8号）
全国名老中医药专家付义传承工作室建设项目(国中医药人教函[2022]75号) （国中医药人教函[2022]75号）
