芪地糖肾方对糖尿病肾病足细胞焦亡及MAPK14/RELA/Caspase-8信号通路的影响  

Implication of Qidi Tangshen Prescription(QDTS)on Podocyte Pyroptosis in Diabetes Nephropathy by Regulating MAPK14/RELA/Caspase-8 Signaling Pathway

作　　者：高飞 于博睿[2] 谢惠迪 周盈 史扬 张先慧 柳红芳[2] GAO Fei;YU Borui;XIE Huidi;ZHOU Ying;SHI Yang;ZHANG Xianhui;LIU Hongfang(Beijing Chaoyang District Hospital of Traditional Chinese Medicine,Beijing 100020,China;Dongzhimen Hospital,Beijing University of Chinese Medicine,Beijing 100700,China;General Hospital of the People's Liberation Army of China,Beijing 100853,China;Beijing Puren Hospital,Beijing 100062,China)

机构地区：[1]北京市朝阳区中医医院,北京100020 [2]北京中医药大学东直门医院,北京100700 [3]中国人民解放军总医院,北京100853 [4]北京市普仁医院,北京100062

出　　处：《中国实验方剂学杂志》2024年第13期67-75,共9页Chinese Journal of Experimental Traditional Medical Formulae

基　　金：国家自然科学基金项目(81774273);北京市自然科学基金项目(7212180);2022年东直门医院科技创新项目(DZMKJCX-2022-003);2023年东直门医院科技创新项目(DZMKJCX-2023-005);北京中医药大学“揭榜挂帅”项目(2023-JYBJBZD-023)。

摘　　要：目的:探讨芪地糖肾方(QDTS)调控糖尿病肾病(DN)足细胞焦亡的分子机制。方法:体内实验将db/db小鼠分为模型组、芪地糖肾方(3.34 g·kg^(-1))和缬沙坦胶囊(10.29 mg·kg^(-1))组,db/m小鼠作为正常组,每组8只,连续干预8周。末次给药后处死小鼠,观察小鼠肾脏病理变化及焦亡活性指标NOD样受体蛋白3(NLRP3)、Gasdermin D蛋白(GSDMD)和白细胞介素-1β(IL-1β)蛋白表达水平。体外实验将小鼠足细胞分为正常糖组(5.5 mmol·L^(-1)葡萄糖),高糖组(35 mmol·L^(-1)葡萄糖),二甲基亚砜(DMSO)组(35 mmol·L^(-1)葡萄糖+200 mg·L^(-1)DMSO)和QDTS组(35 mmol·L^(-1)葡萄糖+200 mg·L^(-1)芪地糖肾方冻干粉),干预48 h后检测足细胞中NLRP3、GSDMD和IL-1β蛋白的表达水平。使用网络药理学方法构建QDTS治疗DN的药物-成分-靶标-疾病交互网络,分析其调控足细胞焦亡的关键信号通路并在体内、体外实验进行验证。结果:与正常组比较,模型组小鼠肾小球肥大,肾小球基底膜增厚,部分节段伴有明显足细胞足突融合,小鼠肾脏中NLRP3、GSDMD和IL-1β蛋白表达升高,小鼠肾脏中丝裂原活化蛋白激酶14(MAPK14)、V-Rel网状内皮增生病毒癌基因同源物A(RELA)和胱天蛋白酶-8(Caspase-8)蛋白表达升高(P<0.05);与模型组比较,QDTS组小鼠肾脏病理损伤明显减轻,QDTS组和缬沙坦组小鼠肾脏中NLRP3、GSDMD和IL-1β蛋白表达降低(P<0.05),QDTS组和缬沙坦组小鼠肾脏中MAPK14、RELA、Caspase-8蛋白表达降低(P<0.05)。网络药理学结果显示QDTS调控DN细胞焦亡的靶点有16个,其中MAPK14、RELA和Caspase-8是关键靶点。与正常糖组比较,高糖组小鼠足细胞中NLRP3、GSDMD和IL-1β蛋白表达升高(P<0.05),小鼠足细胞中MAPK14、RELA和Caspase-8蛋白表达升高(P<0.05);与高糖组比较,QDTS组小鼠足细胞中NLRP3、GSDMD和IL-1β蛋白表达降低(P<0.05),QDTS组小鼠足细胞中MAPK14、RELA和Caspase-8蛋白表达降低(P<0.05)。结论:QDTS减轻DN足细胞�Objective:To explore the molecular mechanism of Qidi Tangshen prescription(QDTS)in regulating podocyte pyroptosis in diabetes nephropathy(DN).Method:Through in vivo experiment,db/db mice were divided into the model group,QDTS group(3.34 g·kg^(-1)),valsartan capsule group(10.29 mg·kg^(-1)),with db/m mice serving as the normal control.Each group consisted of 8 mice,and they underwent continuous intervention for 8 weeks.After the last administration,mice were euthanized,and kidney pathological changes were observed.Additionally,the expression levels of pyroptosis-related indicators,including NOD-like receptor protein 3(NLRP3),Gasdermin D protein(GSDMD),and interleukin-1β(IL-1β)protein,were examined.Through in vitro experiment,mouse podocytes were divided into the normal glucose group(5.5 mmol·L^(-1)glucose),high glucose group(35 mmol·L^(-1)glucose),DMSO group(35 mmol·L^(-1)glucose+200 mg·L^(-1)DMSO),and QDTS group(35 mmol·L^(-1)glucose+200 mg·L^(-1)QDTS freeze-dried powder).After 48 hours of intervention,the expression levels of NLRP3,GSDMD,and IL^(-1)βproteins were measured in podocytes.A drug-ingredient-target-disease interaction network for QDTS in the treatment of DN was constructed by network pharmacology methods.The key signaling pathways regulating podocyte pyroptosis were analyzed,and validation was conducted through in vivo and in vitro experiments.Result:Compared with normal group,glomerular hyperplasia and glomerular basement membrane thickening were observed in model group,and some segments were accompanied by obvious podocellular process fusion.The protein expressions of NLRP3,GSDMD and IL-1βin mouse kidney were increased,the protein expressions of mitogen-activated protein kinase 14(MAPK14),V-Rel reticuloendotheliosis virus oncogene homology A(RELA)and Caspase-8 in mouse kidney were increased(P<0.05).Compared with model group,kidney pathological injury of mice in QDTS group was significantly reduced,and the expressions of NLRP3,GSDMD and IL-1βin kidney of mice in QDTS group and valsartan g

关 键 词：芪地糖肾方 糖尿病肾病 足细胞损伤 焦亡 分子机制 

分 类 号：R2-0[医药卫生—中医学] R33R289R587.1