切换至 "中华医学电子期刊资源库"

中华肾病研究电子杂志 ›› 2019, Vol. 08 ›› Issue (05) : 213 -218. doi: 10.3877/cma.j.issn.2095-3216.2019.05.005

所属专题: 文献

论著

自噬和ASPPs在老年大鼠急性肾损伤早期的表达
李青霖1, 王小丹2, 傅博3, 周飞虎4,()   
  1. 1. 100853 北京,解放军总医院第一医学中心重症医学科
    2. 100853 解放军总医院第二医学中心保健科
    3. 100853 肾脏疾病国家重点实验室
    4. 100853 北京,解放军总医院第一医学中心重症医学科;100853 肾脏疾病国家重点实验室
  • 收稿日期:2019-01-08 出版日期:2019-10-28
  • 通信作者: 周飞虎
  • 基金资助:
    国家老年疾病临床医学研究中心课题(NCRCG-PLAGH-2017008); 吴阶平医学基金会临床科研专项资助(HRJJ20171039/320.6750.18383); 北京市科技计划课题(Z161100000116054)

Expression of autophagy and ASPPs in early acute kidney injury of aged rats

Qinglin Li1, Xiaodan Wang2, Bo Fu3, Feihu Zhou4,()   

  1. 1. Department of Critical Care Medicine, The First Medical Centre
    2. Department of Health Care, The Second Medical Centre
    3. State Key Laboratory of Kidney Diseases; Chinese PLA General Hospital, Beijing 100853, China
    4. Department of Critical Care Medicine, The First Medical Centre; State Key Laboratory of Kidney Diseases; Chinese PLA General Hospital, Beijing 100853, China
  • Received:2019-01-08 Published:2019-10-28
  • Corresponding author: Feihu Zhou
  • About author:
    Corresponding author: Zhou Feihu, Email:
引用本文:

李青霖, 王小丹, 傅博, 周飞虎. 自噬和ASPPs在老年大鼠急性肾损伤早期的表达[J]. 中华肾病研究电子杂志, 2019, 08(05): 213-218.

Qinglin Li, Xiaodan Wang, Bo Fu, Feihu Zhou. Expression of autophagy and ASPPs in early acute kidney injury of aged rats[J]. Chinese Journal of Kidney Disease Investigation(Electronic Edition), 2019, 08(05): 213-218.

目的

观察自噬相关蛋白和p53凋亡刺激蛋白(ASPPs)在肾损伤早期的表达变化,初步探讨自噬相关蛋白和ASPPs是否可能成为老年大鼠AKI早期生物标志物。

方法

建立顺铂致AKI青年与老年大鼠模型。雄性SD老年大鼠随机分为假手术组(Sham),顺铂模型组,同时设数量匹配的雄性SD青年大鼠为对照;模型组大鼠一次性腹腔注射顺铂4 mg/kg,Sham组相同途径注射生理盐水4 ml/kg;在给药12 h、1 d、3 d、5 d、7 d时检测大鼠Scr、BUN;光镜观察大鼠肾脏病理变化;透射电镜观察大鼠肾小管上皮细胞超微结构变化及自噬体的情况;免疫印迹法检测肾脏组织Beclin 1、溶酶体相关膜蛋白2(LAMP2)、p62、p53及ASPP抑制物(iASPP)和ASPP1表达情况。

结果

顺铂诱导12 h后,与Sham组比较,青年与老年大鼠Scr无明显变化(P>0.05);电镜观察到大鼠肾小管上皮细胞自噬体出现而且数量显著增多;老年大鼠肾组织Beclin 1、p62、LAMP-2和p53表达水平明显升高(P<0.05),iASPP表达水平明显降低(P<0.05),并且老年大鼠肾组织Beclin 1、LAMP-2和p53变化时间早于青年大鼠(P<0.05)。

结论

自噬和ASPPs在老年大鼠AKI发生早期即可出现,在Scr开始升高前,反应性自噬已经启动。自噬相关蛋白和ASPPs有望成为AKI早期的损伤标志物,可能是AKI早期干预的新靶点,但仍需更深入的研究。

Objective

To observe the expression changes of autophagy-related proteins and apoptosis-stimulating proteins of p53 (ASPPs) in early renal injury, and to explore whether autophagy-related proteins and ASPPs may become early biomarkers of acute kidney injury (AKI) in aged rats.

Methods

A cisplatin-induced AKI model was established in both young and old rats. Male SD rats were randomly divided into sham operation group and cisplatin model group, and a matched number of male young SD rats were as controls. Rats in the model group were given intraperitoneal injection of cisplatin 4 ml/kg, while rats in the sham group injected with normal saline 4 mg/kg. Rats were examined for Scr and BUN at 12 h, 1 day, 3 days, 5 days, and 7 days, and the pathological changes of rat kidney were observed by light microscopy. Ultrastructural changes and autophagosomes in rat renal tubular epithelial cells were observed under transmission electron microscopy. Western blotting was used to detect the expression of Beclin 1, LAMP-2, p62, p53, iASPP and ASPP1 in kidney tissues.

Results

After 12 hours of cisplatin induction, compared with the sham group, there was no significant change in Scr of young and old rats (P>0.05), and electron microscopy showed that autophagosomes in rat renal tubular epithelial cells appeared and the number increased significantly. The expression levels of Beclin 1, p62, LAMP-2, and p53 in the kidney of aged rats were significantly increased (P<0.05), with the expression level of iASPP being significantly decreased (P<0.05). And the renal expression of Beclin 1, LAMP-2, and p53 changed earlier in the old rats than in the young rats (P<0.05).

Conclusions

Autophagy and ASPPs appeared in the early stage of rat AKI, and reactive autophagy had started before Scr began to rise. Autophagy-associated proteins and ASPPs are expected to be earlier markers of AKI, and may be new targets for early intervention of AKI, which still need further research.

表1 各组大鼠血清Scr和BUN的变化
图1 各组青年与老年大鼠肾组织PAS染色图片( 400×)
图2 青年与老年大鼠肾小管上皮细胞自噬体电镜图片(50 000×)
图3 各组青年与老年大鼠平均出现的自噬体(个)
图4 不同观察时间青年与老年大鼠肾脏组织自噬相关蛋白表达变化
[1]
Anderson S, Eldadah B, Halter JB, et al. Acute kidney injury in older adults [J]. J Am Soc Nephrol, 2011, 22(1): 28-38.
[2]
Bouchard J, Soroko SB, Chertow GM, et al. Fluid accumulation, survival and recovery of kidney function in critically ill patients with acute kidney injury [J]. Kidney Int, 2009, 76(4): 422-427.
[3]
Chao C-T, Tsai H-B, Lin Y-F, et al. Acute kidney injury in the elderly: only the tip of the iceberg [J]. J Clin Gerontol Geriatr, 2014, 5(1): 7-12.
[4]
Iyngkaran P, Schneider H, Devarajan P, et al. Cardio-renal syndrome: new perspective in diagnostics [J]. Semin Nephrol, 2012, 32(1): 3-17.
[5]
程庆砾. 老年人急性肾损伤[J]. 临床肾脏病杂志,2015, 15(7):388-391.
[6]
李青霖,王小丹,杜婧,等. 自噬和p53凋亡刺激蛋白在大鼠急性肾损伤模型中的表达及早期诊断价值[J/CD]. 中华肾病研究电子杂志,2017, 6(3):120-126.
[7]
Mizushima N, Yoshimori T, Levine B. Methods in mammalian autophagy research [J]. Cell, 2010, 140(3): 313-326.
[8]
Salminen A, Kaarniranta K. Regulation of the aging process by autophagy [J]. Trends Mol Med, 2009, 15(5): 217-224.
[9]
Bolignano D, Mattace-Raso F, Sijbrands EJ, et al. The aging kidney revisited: a systematic review [J]. Ageing Res Rev, 2014, 14: 65-80.
[10]
孙雪峰. 老年人急性肾损伤特点[J].中国血液净化,2010, 9(3):123-125.
[11]
Crotzer VL, Blum JS. Autophagy and adaptive immunity [J]. Immunology, 2010, 131(1): 9-17.
[12]
Takabatake Y, Kimura T, Takahashi A, et al. Autophagy and the kidney: health and disease [J]. Nephrol Dial Transplant, 2014, 29(9): 1639-1647.
[13]
Shibutani ST, Yoshimori T. A current perspective of autophagosome biogenesis [J]. Cell Res, 2014, 24(1): 58-68.
[14]
Mehrpour M, Esclatine A, Beau I, et al. Overview of macroautophagy regulation in mammalian cells [J]. Cell Res, 2010, 20(7): 748-762.
[15]
Klionsky DJ, Abdalla FC, Abeliovich H, et al. Guidelines for the use and interpretation of assays for monitoring autophagy [J]. Autophagy, 2012, 8(4): 445-544.
[16]
Lenoir O, Tharaux PL, Huber TB. Autophagy in kidney disease and aging: lessons from rodent models [J]. Kidney Int, 2016, 90(5): 950-964.
[17]
Rubinsztein DC, Marino G, Kroemer G. Autophagy and aging [J]. Cell, 2011, 146(5): 682-695.
[18]
Jiang M, Wei Q, Dong G, et al. Autophagy in proximal tubules protects against acute kidney injury [J]. Kidney Int, 2012, 82(12): 1271-1283.
[19]
Cao Y, Klionsky DJ. Physiological functions of Atg6/Beclin 1: a unique autophagy-related protein [J]. Cell Res, 2007, 17(10): 839-849.
[20]
Xiong J, Xia M, Xu M, et al. Autophagy maturation associated with CD38-mediated regulation of lysosome function in mouse glomerular podocytes [J]. J Cell Mol Med, 2013, 17(12): 1598-1607.
[21]
Nezis IP, Stenmark H. p62 at the interface of autophagy, oxidative stress signaling, and cancer [J]. Antioxid Redox Signal, 2012, 17(5): 786-793.
[22]
Maiuri MC, Galluzzi L, Morselli E, et al. Autophagy regulation by p53 [J]. Curr Opin Cell Biol, 2010, 22(2): 181-185.
[23]
López-Otín C, Blasco MA, Partridge L, et al. The hallmarks of aging [J]. Cell, 2013, 153(6): 1194-1217.
[24]
Wang Y, Godin-Heymann N, Dan Wang X, et al. ASPP1 and ASPP2 bind active RAS, potentiate RAS signalling and enhance p53 activity in cancer cells [J]. Cell Death Differ, 2013, 20(4): 525-534.
[25]
Wang Y, Wang XD, Lapi E, et al. Autophagic activity dictates the cellular response to oncogenic RAS [J]. Proc Natl Acad Sci USA, 2012, 109(33): 13325-13330.
[26]
Wilson AM, Morquette B, Abdouh M, et al. ASPP1/2 regulate p53-dependent death of retinal ganglion cells through PUMA and Fas/CD95 activation in vivo [J]. J Neurosci, 2013, 33(5): 2205-2216.
[27]
Eskelinen EL. Roles of LAMP-1 and LAMP-2 in lysosome biogenesis and autophagy [J]. Mol Aspects Med, 2006, 27(5): 495-502.
[1] 孔莹莹, 谢璐涛, 卢晓驰, 徐杰丰, 周光居, 张茂. 丁酸钠对猪心脏骤停复苏后心脑损伤的保护作用及机制研究[J]. 中华危重症医学杂志(电子版), 2023, 16(05): 355-362.
[2] 韩圣瑾, 周正武, 翁云龙, 黄鑫. 碳酸氢钠林格液联合连续性肾脏替代疗法对创伤合并急性肾损伤患者炎症水平及肾功能的影响[J]. 中华危重症医学杂志(电子版), 2023, 16(05): 376-381.
[3] 张秋彬, 张楠, 林清婷, 徐军, 朱华栋, 姜辉. 急性胰腺炎合并急性肾损伤患者的预后评估[J]. 中华危重症医学杂志(电子版), 2023, 16(05): 382-389.
[4] 罗丹, 孔为民, 陈姝宁, 赵小玲, 谢云凯. 子宫内膜异位症患者在位及异位内膜上皮细胞-间充质转化相关生物标志物的变化[J]. 中华妇幼临床医学杂志(电子版), 2023, 19(05): 530-539.
[5] 代雯荣, 赵丽娟, 李智慧. 细胞外囊泡对胚胎着床影响的研究进展[J]. 中华妇幼临床医学杂志(电子版), 2023, 19(05): 616-620.
[6] 周子慧, 李恭驰, 李炳辉, 王知, 刘慧真, 王卉, 邹利军. 细胞自噬在创面愈合中作用的研究进展[J]. 中华损伤与修复杂志(电子版), 2023, 18(06): 542-546.
[7] 李越洲, 张孔玺, 李小红, 商中华. 基于生物信息学分析胃癌中PUM的预后意义[J]. 中华普通外科学文献(电子版), 2023, 17(06): 426-432.
[8] 陈安, 冯娟, 杨振宇, 杜锡林, 柏强善, 阴继凯, 臧莉, 鲁建国. 基于生物信息学分析CCN4在肝细胞癌中表达及其临床意义[J]. 中华肝脏外科手术学电子杂志, 2023, 12(06): 702-707.
[9] 李青霖, 宋仁杰, 周飞虎. 一种重型劳力性热射病相关急性肾损伤小鼠模型的建立与探讨[J]. 中华肾病研究电子杂志, 2023, 12(05): 265-270.
[10] 任加发, 邬步云, 邢昌赢, 毛慧娟. 2022年急性肾损伤领域基础与临床研究进展[J]. 中华肾病研究电子杂志, 2023, 12(05): 276-281.
[11] 李金璞, 饶向荣. 抗病毒药物和急性肾损伤[J]. 中华肾病研究电子杂志, 2023, 12(05): 287-290.
[12] 王小红, 钱晶, 翁文俊, 周国雄, 朱顺星, 祁小鸣, 刘春, 王萍, 沈伟, 程睿智, 秦璟灏. 巯基丙酮酸硫基转移酶调控核因子κB信号介导自噬对重症急性胰腺炎大鼠的影响及机制[J]. 中华消化病与影像杂志(电子版), 2023, 13(06): 422-426.
[13] 吴蓉菊, 向平超. COPD频繁急性加重表型与炎性因子相关性研究[J]. 中华临床医师杂志(电子版), 2023, 17(9): 939-947.
[14] 易成, 韦伟, 赵宇亮. 急性肾脏病的概念沿革[J]. 中华临床医师杂志(电子版), 2023, 17(08): 906-910.
[15] 邱甜, 杨苗娟, 胡波, 郭毅, 何奕涛. 亚低温治疗脑梗死机制的研究进展[J]. 中华脑血管病杂志(电子版), 2023, 17(05): 518-521.
阅读次数
全文


摘要