IP_(3)R-Grp75-VDAC1-MCU钙轴分子高表达伴mTORC1过度活化参与阿霉素大鼠肾病模型蛋白尿发生  

Involvement of over-production of IP3R-Grp75-VDAC1-MCU calcium axis molecules accompanied with mTORC1 hyper-activation in Adriamycin induced nephropathy rat

作　　者：王斯斯 管娜[1] 许晗 魏骐骄 陶迎红[2] 杨国生[2] Wang Sisi;Guan Na;Xu Han;Wei Qijiao;Tao Yinghong;Yang Guosheng(Department of Pediatrics,Peking University First Hospital,Beijing 100034,China;Experimental Animal Center,Peking University First Hospital,Beijing 100034,China)

机构地区：[1]北京大学第一医院儿科,100034 [2]北京大学第一医院实验动物中心,100034

出　　处：《中华实用儿科临床杂志》2021年第10期767-771,共5页Chinese Journal of Applied Clinical Pediatrics

基　　金：国家自然科学基金(81570639)。

摘　　要：目的分析1,4,5三磷酸肌醇受体(IP_(3)R)-葡萄糖调节蛋白75(Grp75)-电压依赖性阴离子通道1(VDAC1)-线粒体钙单向转运体(MCU)钙轴分子在肾病蛋白尿时表达变化,探讨其上游调控路径。方法将SD大鼠分为对照组(6只)和阿霉素(ADR)组(10只)。通过尾静脉1次注射ADR法建立肾病模型。采用免疫组织化学染色分析肾小球钙轴分子表达和哺乳动物雷帕霉素靶蛋白复合物1(mTORC1)活化标志物。体外培养小鼠足细胞,用ADR诱导足细胞损伤,分别予不同浓度依维莫司干预,分析钙轴分子和凋亡标志物。结果与对照组比较,ADR组大鼠蛋白尿时肾小球IP3R(0.02±0比0)、Grp75(0.04±0比0)、VDAC1(0.04±0比0.01±0)、MCU(0.05±0.01比0.01±0)表达显著增强,mTORC1活化标志物增强(0.57±0.01比0.18±0),差异均有统计学意义(均P<0.001)。在小鼠足细胞中,与对照组比较,ADR组Grp75(1.89±1.17比0.16±0.08)、VDAC1(1.59±0.34比0.20±0.07)、MCU(1.56±0.38比0.46±0.35)表达显著增强,mTORC1活化标志物增强(2.12±0.08比0.39±0.09),差异均有统计学意义(均P<0.05)。与ADR组比较,ADR+依维莫司(1.0 nmol/L)组足细胞Grp75(0.26±0.20比1.89±1.17,P=0.001)、VDAC1(0.40±0.26比1.59±0.34,P=0.014)、MCU(0.60±0.32比1.56±0.38,P=0.029)表达显著减少,线粒体钙显著降低[(2664.00±140.57)U比(3025.16±180.92)U,P=0.023],凋亡标志物活化半胱氨酸天冬氨酸蛋白酶-3显著减少(0.55±0.28比1.48±0.45,P=0.011),差异均有统计学意义。结论IP_(3)R-Grp75-VDAC1-MCU钙轴分子高表达伴mTORC1过度活化参与ADR大鼠肾病模型蛋白尿发生,mTORC1抑制剂显著抑制足细胞钙轴分子表达,可能参与mTORC1抑制剂所致足细胞效应机制。Objective To investigate the expression of inositol 1,4,5-triphate receptor(IP3R)-glucose-regulated protein 75(Grp75)-voltage dependent anion channel 1(VDAC1)-mitochondrial Calcium uniporter(MCU)Calcium axis molecules in proteinuria and to explore its upstream regulator.Methods Sixteen Sprague Dawley rats were divided into control group(6 rats)and Adriamycin(ADR)group(10 rats).Nephropathy rat model was established by single injection of ADR through tail vein.The glomerular expression of IP3R,Grp75,VDAC1,MCU and the activation marker of mammalian target of Rapamycin complex 1(mTORC1)were analyzed by immunohistochemical staining.In cultured mouse podocyte,ADR was used to induce podocyte injury,and the Everolimus of different concentrations was applied for intervention.The expression of the Calcium axis molecules and apoptosis marker was analyzed.Results Compared with control group,the glomerular expression of IP3R(0.02±0 vs.0,P<0.001),Grp75(0.04±0 vs.0,P<0.001),VDAC1(0.04±0 vs.0.01±0,P<0.001),and MCU(0.05±0.01 vs.0.01±0,P<0.001)were significantly increased in ADR-induced nephropathy rats,and the activation marker of mTORC1(0.57±0.01 vs.0.18±0,P<0.001)was increased as well.In cultured mouse podocytes,compared with control group,the expression of Grp75(1.89±1.17 vs.0.16±0.08,P=0.001),VDAC1(1.59±0.34 vs.0.20±0.07,P=0.006),and MCU(1.56±0.38 vs.0.46±0.35,P=0.014)were obviously increased in ADR induced podocytes,and the activation marker of mTORC1(2.12±0.08 vs.0.39±0.09,P<0.001)was also increased.Compared with the ADR induced podocytes,the expression of Grp75(0.26±0.20 vs.1.89±1.17,P=0.001),VDAC1(0.40±0.26 vs.1.59±0.34,P=0.014)and MCU(0.60±0.32 vs.1.56±0.38,P=0.029)in podocytes treated with ADR and 1.0 nmol/L Everolimus were remarkably decreased,accompanied with the decrease of mitochondrial calcium[(2664.00±140.57)U vs.(3025.16±180.92)U,P=0.023]and apoptosis marker cleaved Caspase-3(0.55±0.28 vs.1.48±0.45,P=0.011).Conclusions The over-production of IP3R-Grp75-VDAC1-MCU Calcium axis molecules ac

关 键 词：蛋白尿 足细胞 哺乳动物雷帕霉素靶蛋白复合物1 

分 类 号：R965[医药卫生—药理学]