PKD1抑制剂CID755673通过诱导线粒体功能障碍加重急性肾损伤  

PKD1 inhibitor CID755673 aggravates acute kidney injury by inducing mitochondrial dysfunction

作　　者：郭燕 徐爽 王亭 GUO Yan;XU Shuang;WANG Ting(Pediatric Hospital of Nanjing Medical University/Key Laboratory of Research on Children’s Major Diseases,Nanjing 210029,China;Department of Pediatrics,the Sir Run Run Hospital Affiliated to Nanjing Medical University,Nanjing 211112,China)

机构地区：[1]南京医科大学儿科学院/江苏省儿童重大疾病研究重点实验室,南京210029 [2]南京医科大学附属逸夫医院儿科,南京211112

出　　处：《临床与实验病理学杂志》2023年第2期206-211,215,共7页Chinese Journal of Clinical and Experimental Pathology

基　　金：国家自然科学青年基金(81800656)。

摘　　要：目的 探讨PKD1在急性肾损伤(acute kidney injury, AKI)中的表达及其作用机制。方法 采用免疫组化法检测癌旁正常肾组织和AKI患者肾活检组织中以及顺铂(Cisplatin)处理的小鼠肾组织中PKD1的表达;体外培养小鼠肾小管上皮细胞(mouse proximal tubular epithelial cells, mPTC),应用qRT-PCR和Western blot法检测PKD1及凋亡相关基因mRNA和蛋白表达水平;运用流式细胞术检测细胞凋亡、线粒体膜电位及线粒体活性氧水平。结果 免疫组化检测:PKD1在AKI患者肾活检组织中的表达(4.07±0.994)明显高于癌旁正常肾组织(1.001±0.073,P=0.021 1);在Cisplatin诱导的小鼠AKI肾组织中也发现,与对照组(0.992±0.072)相比,AKI肾组织中PKD1表达水平明显上调(2.457±0.317,P=0.002 5);在mPTC中,与Cisplatin组比较,Cisplatin+CID组中Bax mRNA及蛋白表达增加(P<0.05)及Caspase-3活化水平升高(P=0.026),抗凋亡蛋白BCL-2 mRNA及蛋白水平则进一步下调(P<0.05)。流式细胞术实验:与Cisplatin组比较,Cisplatin+CID组细胞凋亡百分比(Cisplatin组为21.77±2.378、Cisplatin+CID组为43.01±2.796,P<0.000 1)及线粒体活性氧水平(Cisplatin组为1.562±0.023、Cisplatin+CID组为1.957±0.088,P=0.002)进一步增加;而线粒体膜电位水平进一步降低(Cisplatin组为0.725 7±0.163、Cisplatin+CID组为0.46±0.026,P=0.043)。结论 PKD1在AKI损伤中显著上调,抑制PKD1明显加重Cisplatin诱导的AKI,其有望成为防治AKI的新靶点。Purpose To investigate the expression and mechanism of PKD1 in acute kidney injury(AKI). Methods Immunohistochemistry was used to detect the expression of PKD1 in normal tissues near the cancer of the kidney, kidney tissues of AKI patients and kidney tissues of Cisplatin-treated mice. Mouse proximal tubular epithelial cells(mPTC) were cultured in vitro, and the mRNA and protein expression levels of PKD1 and apoptosis related genes were detected by qRT-PCR and Western blot. Cell apoptosis, mitochondrial membrane potential and mitochondrial reactive oxygen species were detected by flow cytometry. Results Immunohistochemical test showed that the expression of PKD1 in pathological section of AKI patients(4.07±0.994) was significantly higher than that in adjacent normal tissue(1.001±0.073, P=0.0211), and which was also higher in Cisplatin-induced mouse kidney tissues. Compared with the normal group(0.992±0.072), the level of PKD1 in AKI kidney tissue was significantly up-regulated(2.457±0.317, P=0.0025). In mPTC, mRNA and protein of BAX were increased in Cisplatin+CID group compared with Cisplatin group(P<0.05) and the activation level of Caspase-3 was increased(P=0.026), and the mRNA and protein levels of anti-apoptotic protein BCL-2 were further down-regulated(P<0.05). Compared with Cisplatin group, the percentage of apoptosis in Cisplatin group(21.77±2.378) and Cisplatin+CID group(43.01±2.796 was also tested by flow cytometry, P<0.0001) and mitochondrial reactive oxygen species(Cisplatin group: 1.562±0.023, Cisplatin+CID group: 1.957±0.088, P=0.002) were further increased. Mitochondrial membrane potential was further decreased(Cisplatin group: 0.725 7±0.163, Cisplatin+CID group: 0.46±0.026, P=0.043). Conclusion PKD1 is significantly up-regulated in AKI injury, and inhibition of PKD1 can significantly aggravate Cisplatin-induced AKI, which is expected to be a new target for the prevention and treatment of AKI.

关 键 词：肾损伤 AKI PKD1 线粒体功能障碍 CISPLATIN 

分 类 号：R363.2[医药卫生—病理学] R736.6[医药卫生—基础医学]