TDO2介导肾小管上皮细胞凋亡在Cis-AKI中作用及机制  

Role and mechanism of TDO2 mediated apoptosis ofrenal tubular epithelial cells in Cis-AKI

作　　者：林茜茜 宗雪梅 陈悦兰 汪文莉 王越业 严尚学[1,2] 魏伟[1] 常艳[1,2] LIN Qian-qian;ZONG Xue-mei;CHEN Yue-lan;WANG Wen-li;WANG Yue-ye;YAN Shang-xue;WEI Wei;CHANG Yan(Clinical Pharmacology Research Institute of Anhui Medical University,Key Laboratory of Anti-inflammatory and Immune Drugs of the Ministry of Education,Anhui Anti-inflammatory and Immune Drug Collaborative Innovation Center,Rheumatoid Arthritis Research Center of Anhui Medical University,Hefei 230032,China;Experimental Animal Center of Anhui Medical University,Hefei 230032,China)

机构地区：[1]安徽医科大学临床药理研究所,抗炎免疫药物教育部重点实验室,安徽省抗炎免疫药物协同创新中心,安徽医科大学类风湿关节炎研究中心 [2]安徽医科大学实验动物中心,安徽合肥230032

出　　处：《中国药理学通报》2025年第3期475-482,共8页Chinese Pharmacological Bulletin

基　　金：安徽省自然科学基金面上项目(No 2108085MH320);省留学人员创新项目择优资助计划项目(No 200LCX019);2024年度安徽省高校年度科研计划项目(No 2024AH050745);安徽省转化医学研究院科研基金项目(No 2023zhyx-C11)。

摘　　要：目的探讨色氨酸-2,3-双加氧酶(tryptophan 2,3-dioxygenase,TDO2)在顺铂诱导急性肾损伤(cisplatin-acute kidney injury,Cis-AKI)中的作用,并在肾小管上皮细胞(tubular epithelial cells,TECs)中探究TDO2与凋亡相关机制。方法采用腹腔注射顺铂(cisplatin,Cis)建立AKI模型。比色法检测CRE、BUN水平,PAS染色观察小鼠肾脏损伤。免疫组化检测TDO2蛋白表达和分布及巨噬细胞(F4/80+)浸润;免疫荧光检测TDO2与肾小管标记物LTL共定位;TUNEL染色检测小鼠肾脏细胞凋亡;流式细胞术检测人肾皮质近曲小管上皮细胞(HK2)过表达和给予TDO2抑制剂680C91后细胞凋亡情况;Western blot检测过表达和抑制TDO2后HK2细胞TDO2、NF-κB信号通路蛋白水平。结果在整体动物实验中,与对照组相比,Cis-AKI小鼠CRE、BUN水平均明显升高,肾小管损伤明显;同时,Cis-AKI小鼠肾脏组织F4/80表达增加且肾脏细胞凋亡比例增加。免疫组化和免疫荧光结果表明TDO2表达增加且主要定位在TECs中。在细胞实验中,HK2细胞过表达TDO2后凋亡比例增加,TDO2、p-IκBα、p-p65蛋白表达升高,p-IκBα/IκBα、p-p65/p65升高;此外,给予680C91后细胞凋亡比例减少,p-IκBα、p-p65蛋白表达降低,p-IκBα/IκBα、p-p65/p65降低。结论TECs中TDO2升高参与Cis-AKI病理机制,可能与其诱导TECs凋亡和激活NF-κB信号通路进而参与肾损伤有关。Aim To investigate the role of tryptophan 2,3-dioxygenase(TDO2)in cisplatin-acute kidney injury(Cis-AKI)and to explore the mechanism of TDO2 in relation to apoptosis in tubular epithelial cells(TECs)to investigate the mechanism of TDO2 associated with apoptosis.Methods An AKI model was established by intraperitoneal injection of cisplatin(Cis).Colorimetric assay was used to detect CRE and BUN levels,and PAS staining was employed to observe renal injury in mice.Immunohistochemistry was used to detect TDO2 protein expression and distribution and macrophage(F4/80+)infiltration;immunofluorescence was used to detect the co-localization of TDO2 with the tubular marker LTL;TUNEL staining was used to detect apoptosis in mouse kidney;flow cytometry was used to detect overexpression of human renal cortical proximal tubular epithelial cells(HK2)and apoptosis after administration of the TDO2 inhibitor 680C91;Western blot was used to detect TDO2 and NF-κB pathway protein levels in HK2 cells after over-expression and inhibition of TDO2.Results In the overall animal experiments,Cis-AKI mice showed significantly higher levels of CRE and BUN and obvious tubular damage compared with the control group;at the same time,the renal tissues of Cis-AKI mice showed increased expression of F4/80,and the proportion of apoptotic cells in kidney cells was increased.Immunohistochemistry and immunofluorescence showed that the expression of TDO2 increased,mainly localized in TECs.In cellular experiments,HK2 cells overexpressing TDO2 increased the proportion of apoptosis,and the expression of TDO2,p-IκBα,and p-p65 proteins was elevated,and p-IκBα/IκBαand p-p65/p65 were elevated;furthermore,the proportion of apoptosis was reduced by the administration of 680C91,and the expression of p-IκBα,and p-p65 proteins decreased,and the expression of p-IκBα/IκBα,and p-p65/p65 decreased.Conclusions Elevated TDO2 in TECs is involved in the pathological mechanism of Cis-AKI,which may be related to its induction of apoptosis in TECs and activation o

关 键 词：急性肾损伤 顺铂 HK2 TDO2 凋亡 NF-ΚB 

分 类 号：R-332[医药卫生] R322.61R329.25R345.47R692