薯蓣皂苷通过抑制Akt/GSK-3β信号通路减轻UUO诱导肾纤维化的机制研究OA
Dioscin attenuates UUO-induced renal fibrosis by inhibiting the Akt/GSK-3β signaling pathway
目的:探讨薯蓣皂苷(dioscin,Dis)对单侧输尿管梗阻(unilateral ureteral occlusion,UUO)诱导的肾纤维化及转化生长因子-β1(transforming growth factor-beta 1,TGF-β1)诱导的人肾小管上皮(human renal tubular ep-ithelial cells,HK-2)细胞上皮-间质转化(epithe-lial-mesenchymal transition,EMT)样表型改变的影响及机制.方法:将 C57BL/6J 雄性小鼠随机分为假手术、UUO、UUO+Dis(50 mg/kg、100 mg/kg)组.术后干预 7 d,取肾组织进行苏木精-伊红和Masson 染色观察肾组织病理学改变;免疫组化、蛋白质印迹法(Western blot)和实时荧光定量PCR(real time PCR,RT-PCR)检测蛋白激酶 B(protein kinase B,Akt)/糖原合成酶激酶 3β(glycogen synthase kinase 3β,GSK-3β)通路及EMT 相关分子表达.体外实验以 TGF-β1诱导HK-2细胞 EMT 样表型改变,给予 Dis 或/和 Akt 激动剂 SC79干预 24 h,通过 Western blot、免疫荧光和 RT-PCR 检测上述指标.结果:Dis 显著改善UUO 模型小鼠肾脏病理损伤和胶原沉积,能抑制UUO 模型小鼠肾脏中 Akt/GSK-3β 磷酸化(P<0.05),降低纤维连接蛋白、α-平滑肌肌动蛋白(alpha smooth muscle actin,α-SMA)、波形蛋白(Vimentin)表达(P<0.05),上调 E-钙黏蛋白表达(P<0.05).体外实验中,Dis 抑制 TGF-β1诱导的 Akt/GSK-3β 磷酸化(P<0.01),逆转 α-SMA及 Vimentin 表达升高(P<0.01).此外,SC79可激活该通路并加剧 TGF-β1诱导的 EMT 样表型改变(P<0.05),而 Dis 能逆转 SC79的上述作用(P<0.01).结论:Dis 可能通过抑制 Akt/GSK-3β 信号通路过度激活,减轻 UUO 模型肾纤维化及 TGF-β1诱导的 EMT 样表型改变.
AIM:To investigate the effect of dioscin(Dis)on unilateral ureteral obstruction(UUO)-induced renal fibrosis and on transforming growth factor-β1(TGF-β1)-induced epithelial-mes-enchymal transition(EMT)-like phenotypic changes in human renal tubular epithelial(HK-2)cells,as well as its underlying mechanism.METHODS:Male C57BL/6J mice were randomly divided into sham-operated,UUO,UUO+Dis(50 mg/kg),and UUO+Dis(100 mg/kg)groups.After 7 days of intervention,kidney tissues were collected for hematoxylin-eosin and Masson's trichrome staining to observe histopathological changes.Immunohistochemistry,Western blot,and real-time PCR(RT-PCR)were used to detect the expression of protein kinase B(Akt)/glycogen synthase kinase 3β(GSK-3β)path-way and EMT-related molecules.HK-2 cells were in-duced with TGF-β1 to establish an EMT-like pheno-typic changes model and treated with Dis and/or the Akt agonist SC79 for 24 h.Western blot,im-munofluorescence,and RT-PCR were performed to evaluate the above indicators.RESULTS:Dis signifi-cantly ameliorated renal pathological injury and col-lagen deposition in UUO mice.It inhibited the phos-phorylation of Akt and GSK-3β in UUO kidneys(P<0.05),downregulated the expression of fi-bronectin,α-smooth muscle actin(α-SMA),and Vi-mentin(P<0.05),and upregulated E-cadherin ex-pression(P<0.05).In HK-2 cells,Dis suppressed TGF-β1-induced phosphorylation of Akt/GSK-3β(P<0.01)and reversed the upregulation of α-SMA and Vi-mentin(P<0.01).Furthermore,SC79 activated this pathway and aggravated TGF-β1-induced EMT-like phenotypic changes(P<0.05),whereas Dis reversed these effects of SC79(P<0.01).CONCLUSION:Dis may alleviate UUO-induced renal fibrosis and TGF-β1-induced EMT-like phenotypic changes by inhibit-ing the overactivation of the Akt/GSK-3β signaling pathway.
高骏伟;马贵婕;林晓蒙;邓颖萍;尹程倩;吴宸广;刘鹏;钟光辉
浙江中医药大学附属宁波市中医院,宁波 315010,浙江十堰市中西医结合医院,十堰 442000,湖北浙江中医药大学附属宁波市中医院,宁波 315010,浙江浙江中医药大学附属宁波市中医院,宁波 315010,浙江黑龙江省中医药科学院,哈尔滨 150000,黑龙江黑龙江省中医药科学院,哈尔滨 150000,黑龙江中国中医科学院西苑医院,北京 100091浙江中医药大学附属宁波市中医院,宁波 315010,浙江
医药卫生
肾纤维化UUO模型小鼠薯蓣皂苷Akt/GSK-3β通路SC79
renal fibrosisUUO micedioscin(Dis)Akt/GSK-3β pathwaySC79
《中国临床药理学与治疗学》 2026 (9)
1182-1192,11
宁波市中医肾病临床医学研究中心(2023L001)浙江省自然科学基金联合项目(LHDMY24H270001,LBY24H290002)浙江省中医药重点学科建设项目(2024-XK-63)宁波市中医肾病临床医学研究中心(2024L001)宁波市重点研发计划暨"揭榜挂帅"项目(2023Z165)浙江省中医药科技计划项目(2026ZL0117,2024ZL142)宁波市科技计划项目(2023Z165,2024J259)
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