金芪志颗粒调控ATF-4/NUPR1信号轴抑制软骨细胞铁死亡改善膝骨关节炎的机制OA
Study on the mechanism of Jinqizhi Granule(金芪志颗粒)in regulating the ATF-4/NUPR1 signaling pathway to inhibit ferroptosis in chondrocytes and alleviate knee osteoarthritis
目的 基于ATF-4/NUPR1信号轴探讨金芪志颗粒对膝关节炎(KOA)大鼠软骨细胞铁死亡的影响及其作用机制.方法 将40只SD大鼠,随机分为对照组、KOA组、金芪志颗粒低、中、高剂量组,每组8只.采用半月板失稳术构建KOA大鼠模型.术后2周开始灌胃干预,连续4周.通过番红O-固绿染色观察软骨组织病理变化并进行Mankin评分;Western blot、qPCR及免疫组化法检测软骨组织中ATF-4、NUPR1、铁死亡关键蛋白(ACSL4、GPX4、SLC7A11)及基质代谢指标(MMP-13、Collagen Ⅱ)的表达.结果 与对照组比较,KOA模型组软骨结构破坏严重,Mankin评分显著升高(P<0.01);ATF-4、NUPR1、ACSL4、MMP-13的蛋白及mRNA表达均上调(P<0.01),而GPX4、SLC7A11、Collagen Ⅱ表达下调(P<0.01).金芪志颗粒干预后,软骨损伤明显改善,Mankin评分降低,且呈剂量依赖性下调ATF-4、NUPR1、ACSL4、MMP-13表达,上调GPX4、SLC7A11、Collagen Ⅱ表达(P<0.01).免疫组化结果进一步验证上述蛋白表达趋势.结论 金芪志颗粒可能通过抑制ATF-4/NUPR1信号轴激活,减轻软骨细胞铁死亡,延缓KOA软骨退变,其作用与调控铁死亡相关蛋白ACSL4、GPX4、SLC7A11及改善基质代谢平衡密切相关.
Objective To investigate whether Jinqizhi Granule(金芪志颗粒,JQZG)can inhibit chondrocyte ferroptosis by modulating the ATF-4/NUPR1 signaling pathway in knee osteoarthritis(KOA)rats,thereby protecting cartilage tissue.Methods A KOA rat model was established via meniscal instability surgery.Rats were divided into five groups:control,KOA,and low/medium/high-dose JQZG groups(n=8 per group).The control group underwent sham operation.Two weeks post-operation,low/medium/high-dose groups received oral administration of JQZG at 6.4,12.8,and 25.6 g/(kg·d),respectively.The sham-operated and KOA groups received saline via oral gavage once daily for 4 consecutive weeks.Chondral tissue pathological changes were observed using Fuchsin-Green staining and scored according to the Mankin grading system.Western blot(WB)and qPCR were used to detect chondral tissue markers of the ATF-4/NUPR1 pathway[Activated Transcription Factor 4(ATF-4),Nucleoprotein 1(NUPR1)],ferroptosis markers[acyl-CoA synthase 4(ACSL4),glutathione peroxidase 4(GPX4),solute carrier family 7 member 11(SLC7A11)],and matrix metabolism mark-ers[matrix metalloproteinase-13(MMP-13),type II collagen(Collagen II)].Immunohistochemistry(IHC)examined the expression levels of ATF-4,NUPR1,ACSL4,GPX4,and SLC7A11 in cartilage tissue across all groups.Results Compared with the sham-operated group,rats in the KOA group exhibited severe cartilage margin destruction,elevated Mankin scores,and decreased protein and mRNA expression levels of GPX4,SLC7A11,and Collagen II,while showing increased expression levels of ATF-4,NUPR1,ACSL4,and MMP-13(P<0.01).Compared with the KOA group,all intervention groups exhibited decreased ATF-4,NUPR1,ACSL4,and MMP-13 protein and mRNA expression levels(P<0.01),while GPX4,SLC7A11,and Collagen II protein and mRNA expression levels were increased.IHC analysis of cartilage tissue after intervention showed a significant decrease in ATF-4,NUPR1,ACSL4 in cartilage tissue was significantly reduced,while the proportion of GPX4 and SLC7A11 positive areas was markedly increased.Conclusion JQZG can improve cartilage damage in KOA by regulating the ATF-4/NUPR1 pathway to inhibit ferroptosis in chondrocytes.
匡辰;魏义保;刘德仁;胡恩睿;龚子健;茆军;张农山;廖太阳;王培民
南京中医药大学附属医院,江苏 南京 210004||南京中医药大学昆山附属医院,江苏 昆山 215300||南京中医药大学第一临床医学院,江苏 南京 210004南京中医药大学附属医院,江苏 南京 210004||南京中医药大学第一临床医学院,江苏 南京 210004南京中医药大学附属医院,江苏 南京 210004||南京中医药大学第一临床医学院,江苏 南京 210004南京中医药大学附属医院,江苏 南京 210004||南京中医药大学第一临床医学院,江苏 南京 210004南京中医药大学附属医院,江苏 南京 210004||南京中医药大学第一临床医学院,江苏 南京 210004南京中医药大学附属医院,江苏 南京 210004||南京中医药大学第一临床医学院,江苏 南京 210004南京中医药大学附属医院,江苏 南京 210004||南京中医药大学第一临床医学院,江苏 南京 210004南京中医药大学附属第二医院,江苏 南京 210004南京中医药大学附属医院,江苏 南京 210004||南京中医药大学第一临床医学院,江苏 南京 210004
医药卫生
金芪志颗粒膝骨关节炎软骨退变铁死亡ATF-4/NUPR1信号轴
Jinqizhi Granule(金芪志颗粒)Knee osteoarthritisCartilage degenerationFerroptosisATF-4/NUPR1 Signal-ing pathway
《时珍国医国药》 2026 (11)
2007-2014,8
国家自然科学基金(82305276)江苏省中医药科技发展项目(QN202105)江苏省高等学校自然科学研究项目(22KJB360008)江苏省中医院膝关节骨关节炎临床医学创新中心项目(Y2023zx05)江苏省医学重点学科(实验室)培育单位项目(JSDW202252)苏州市科技发展计划项目(SYSD2020036)苏州市吴门医派研究专项科研基金(SYWD2025270)
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