Early diagnosis value of autophagy and expression of apoptosis-stimulating proteins of p53 in acute kidney injury model of rats

doi:10.3877/cma.j.issn.2095-3216.2017.03.006

Abstract

Abstract:

Objective

To observe the expression changes of autophagy-related proteins and apoptosis-stimulating proteins of p53 (ASPPs) in early kidney injury, in order to investigate whether autophagy-related proteins and ASPPs can be biomarkers of early acute kidney injury (AKI) induced by cisplatin in Sprague-Dawley (SD) rats.

Methods

A total of 46 eight-week-old male SD rats were randomly divided into sham group and cisplatin model group. The rats in the model group were injected intraperitoneally with cisplatin 5mg/kg once, while rats in the sham group were treated in the same way with normal saline of the same volume. Serum creatinine (Scr), blood urea nitrogen (BUN), and cystatin C (Cys C) were measured at 1, 3, 7, 10 and 14 days after the administration. The kidneys pathological changes were observed under light microscopy. Ultrastructural changes and autophagy of rat renal tubular epithelial cells were observed under transmission electron microscopy. Expressions of LC3, Atg5-Atg12, Beclin 1, LAMP-2, p62, p53, iASPP, and ASPP1 were detected with Western blotting.

Results

At 1 day after the administration, levels of Scr, BUN, and Cys C in the cisplatin group were significantly higher than those in the sham group (P<0.05), and the renal tubular injury was significantly increased under light microscopy. The results of electron microscopy showed that the average number of autophagy was significantly increased in the cisplatin model group. From 1 day after the administration, the expression of iASPP protein in kidneys tissues was significantly decreased (P<0.05), while the expressions of Atg5-Atg12, Beclin1, LAMP-1, p62, p53, and ASPP1 proteins in the cisplatin model group was significantly higher than those in the sham group (P<0.05). The expression of LC3 in renal tissues was significantly decreased from 3 days after the administration (P<0.05).

Conclusions

Autophagy and ASPPs occurred in early AKI, and were involved in the development and progression of AKI. Reactive autophagy had been initiated before Scr began to increase. Although autophagy-related proteins and ASPPs are expected to be earlier markers of AKI, which may provide new targets for early intervention in AKI, further studies are still needed.

Key words: Cisplatin, Acute kidney injury, Biomarker, Apoptosis-stimulating proteins of p53, Autophagy

Qinglin Li, Xiaodan Wang, Jing Du, Yuru Li, Bo Fu, Yang Lyu. Early diagnosis value of autophagy and expression of apoptosis-stimulating proteins of p53 in acute kidney injury model of rats[J]. Chinese Journal of Kidney Disease Investigation(Electronic Edition), 2017, 06(03): 120-126.

/ / Recommend

Add to citation manager EndNote|Ris|BibTeX

URL: https://zhsbyjdzzz.cma-cmc.com.cn/EN/10.3877/cma.j.issn.2095-3216.2017.03.006

https://zhsbyjdzzz.cma-cmc.com.cn/EN/Y2017/V06/I03/120

Figures/Tables 5

References 39

[1]	Xu X, Nie S, Liu Z, et al. Epidemiology and clinical correlates of AKI in Chinese hospitalized adults [J]. Clin J Am Soc Nephrol, 2015, 10(9): 1510-1518.
[2]	Kilic U, Kilic E, Tuzcu Z, et al. Melatonin suppresses cisplatin-induced nephrotoxicity via activation of Nrf-2/HO-1 pathway [J]. Nutr Metab (Lond), 2013, 10(1): 7.
[3]	Wei Q, Dong G, Yang T, et al. Activation and involvement of p53 in cisplatin-induced nephrotoxicity [J]. Am J Physiol Renal Physiol, 2007, 293(4): F1282-F1291.
[4]	Kidney Disease: Improving Global Outcomes (KDIGO) Acute Kidney Injury Work Group. KDIGO clinical practice guideline for acute kidney injury [J]. Kidney Int Suppl, 2012, 2: 1-138.
[5]	程庆砾．老年人急性肾损伤[J]. 临床肾脏病杂志，2015，(7): 388-391.
[6]	蔡广研，卜茹．急性肾损伤概念和诊断标准的变迁[J/CD]. 中华肾病研究电子杂志，2013，2(3): 115-119.
[7]	Lynch GS. Emerging drugs for sarcopenia: age-related muscle wasting[J]. Expert Opin Emerg Drugs, 2008, 13(4): 345-361.
[8]	李荣山．急性肾损伤的生物标志物[J/CD]. 中华肾病研究电子杂志，2013，2(3): 134-137.
[9]	Iyngkaran P, Schneider H, Devarajan P, et al. Cardio-renal syndrome: new perspective in diagnostics [J]. Semin Nephrol, 2012, 32(1): 3-17.
[10]	李立斌，严静．急性肾损伤的早期诊治：路在何方？[J]. 中华危重病急救医学，2014，26(4): 209-211.
[11]	Crotzer VL, Blum JS. Autophagy and adaptive immunity [J]. Immunology, 2010, 131(1): 9-17.
[12]	Livingston MJ, Dong Z. Autophagy in acute kidney injury [J]. Semin Nephrol, 2014, 34(1): 17-26.
[13]	Kaushal GP, Shah SV. Autophagy in acute kidney injury [J]. Kidney Int, 2016, 89(4): 779-791.
[14]	Periyasamy-Thandavan S, Jiang M, Wei Q, et al. Autophagy is cytoprotective during cisplatin injury of renal proximal tubular cells [J]. Kidney Int, 2008, 74(5): 631-640.
[15]	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.
[16]	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.
[17]	Song B, Bian Q, Zhang YJ, et al. Downregulation of ASPP2 in pancreatic cancer cells contributes to increased resistance to gemcitabine through autophagy activation [J]. Mol Cancer, 2015, 14: 177.
[18]	Chikh A, Sanza P, Raimondi C, et al. iASPP is a novel autophagy inhibitor in keratinocytes [J]. J Cell Sci, 2014, 127(Pt 14): 3079-3093.
[19]	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.
[20]	肖梦云，王小丹，洪权，等．洛汀新和科素亚对糖尿病肾病大鼠足细胞的保护作用[J]. 西安交通大学学报:医学版，2014，35(5): 659-664.
[21]	潘明娇，王小丹．自噬在高血压足细胞损伤中的作用[J]. 中国循环杂志，2015，30(1): 94-97.
[22]	潘明娇，肖梦云，吕杨，等．贝那普利和氯沙坦对老年自发性高血压大鼠肾小球足细胞自噬的影响及作用机制[J]. 中华老年医学杂志，2016，35(8): 888-893.
[23]	潘明娇，肖梦云，吕杨，等．衰老和高血压对大鼠肾皮质血管紧张素Ⅱ及肾小球足细胞自噬的影响[J]. 中华老年医学杂志，2016，35(4): 421-426.
[24]	Mizushima N, Yoshimori T, Levine B. Methods in mammalian autophagy research [J]. Cell, 2010, 140(3): 313-326.
[25]	Takabatake Y, Kimura T, Takahashi A, et al. Autophagy and the kidney: health and disease [J]. Nephrol Dial Transplant, 2014, 29(9): 1639-1647.
[26]	Shibutani ST, Yoshimori T. A current perspective of autophagosome biogenesis [J]. Cell Res, 2014, 24(1): 58-68.
[27]	Mehrpour M, Esclatine A, Beau I, et al. Overview of macroautophagy regulation in mammalian cells [J]. Cell Res, 2010, 20(7): 748-762.
[28]	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.
[29]	Lenoir O, Tharaux PL, Huber TB. Autophagy in kidney disease and aging: lessons from rodent models [J]. Kidney Int, 2016, 90(5): 950-964.
[30]	Rubinsztein DC, Cuervo AM, Ravikumar B, et al. In search of an "autophagomometer" [J]. Autophagy, 2009, 5(5): 585-589.
[31]	Barth S, Glick D, Macleod KF. Autophagy: assays and artifacts [J]. J Pathol, 2010, 221(2): 117-124.
[32]	Kundu M, Thompson CB. Autophagy: basic principles and relevance to disease [J]. Annu Rev Pathol, 2008, 3: 427-455.
[33]	Weidberg H, Shvets E, Elazar Z. Biogenesis and cargo selectivity of autophagosomes [J]. Annu Rev Biochem, 2011, 80: 125-156.
[34]	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.
[35]	Cao Y, Klionsky DJ. Physiological functions of Atg6/Beclin 1: a unique autophagy-related protein [J]. Cell Research, 2007, 17(10): 839-849.
[36]	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.
[37]	Kimura T, Takabatake Y, Takahashi A, et al. Autophagy protects the proximal tubule from degeneration and acute ischemic injury [J]. J Am Soc Nephrol, 2011, 22(5): 902-913.
[38]	Rovetta F, Stacchiotti A, Consiglio A, et al. ER signaling regulation drives the switch between autophagy and apoptosis in NRK-52E cells exposed to cisplatin [J]. Exp Cell Res, 2012, 318(3): 238-250.
[39]	Yang C, Kaushal V, Shah SV, et al. Autophagy is associated with apoptosis in cisplatin injury to renal tubular epithelial cells [J]. Am J Physiol Renal Physiol, 2008, 294(4): F777-F787.

Please choose a citation manager

Content to export