Extracellular vesicles derived from human umbilical cord mesenchymal stem cells ameliorate acute kidney injury in mice

doi:10.3877/cma.j.issn.2095-1221.2018.05.002

Abstract

Abstract:

Objectives

To explore the effect of Oct-4 overexpressing mesenchymal stem cell derived extracellular vesicles (MSC-EVs) on acute kidney injury (AKI) and the related mechanism.

Methods

The renal tubular epithelial cells treated in low oxygen environment were cocultured with MSC-EVs and control medium for 48 hours. BrdU and TUNEL were used to assess cell proliferation and apoptosis. Mice with acute kidney injury were randomly divided into 4 groups, which were injected with "Vehicle", "EVs", Oct-4 overexpressing EVs (EVs + Oct-4) , and Oct-4 knocked-down EVs (EVs–Oct-4) . Blood creatinine and urine nitrone levels were assessed 48 hours after injection. The mice kedneys were harvested for TUNEL and PCNA staining to evaluate apoptosis and proliferation. Masson trichrome staining was used to evaluate renal fibrosis. Snail gene expression was assessed by PCR. The data was analyzed by ANOVA test and SNK-q test.

Results

48 hours after hypoxic treatment, TUNEL showed the EVs + Oct-4 group has a lowest apoptosis level (0-1) /?HPF. BrdU showed the EVs?+?Oct-4 group has a highest proliferation level (7±2) /HPF. Injection of Oct-4 overexpressing EVs could significantly decrease the level of blood Crea (28?mmol/?L) and BUN (17?mmol/L) , and the kidney fibrosis was also alleviated, as shown by slight Masson staining (3.2±1.0) ﹪ and few PCNA positive cells (70±7) /HPF 48?h post AKI, (21±4) /HPF 2?week post AKI) , the number of TUNEL positive cells was decreased (2±1) /HPF 48?h post AKI, (3±2) /?HPF 2?week post AKI. PCR showed that Vehicle group had a highest Snail level 2.3±0.2, and the EVs?+ Oct-4 group had a lowest Snail level 0.7±0.1.

Conclusions

MSC-EVs provides therapeutic effects in ischemic-reperfusion injury. Oct-4 overexpression could enhance its therapeutic effects by inhibiting apoptosis, promoting proliferation, and inhibiting fibrosis.

Key words: Acute kidney injury, Mesenchymal stem cells, Extracellular vesicles

Zhiyuan Zhang, Yanping Hou, Xiangyu Zou, Jin Zhou, Xiaoyu Xing, Guanqun Ju, Liang Zhong, Jie Sun. Extracellular vesicles derived from human umbilical cord mesenchymal stem cells ameliorate acute kidney injury in mice[J]. Chinese Journal of Cell and Stem Cell(Electronic Edition), 2018, 08(05): 264-271.

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Figures/Tables 8

References 25

1	Bellomo R, Kellum JA, Ronco C. Acute kidney injury[J]. Lancet, 2012, 380(9843):756-766.
2	王万里, 李青山, 周颖, 等. DCD供肝肝移植术后早期急性肾损伤相关危险因素分析[J]. 器官移植, 2018, 9(2):130-136.
3	Zuk A, Bonventre JV. Acute kidney injury[J]. Annu Rev Med, 2016, 67:293-307.
4	Korula S, Balakrishnan S, Sundar S, et al. Acute kidney injury-incidence, prognostic factors, and outcome of patients in an Intensive Care Unit in a tertiary center: A prospective observational study[J]. Indian J Crit Care Med, 2016, 20(6):332-336.
5	Libório AB, Branco KM, Torres de Melo Bezerra C. Acute kidney injury in neonates: from urine output to new biomarkers[J]. Biomed Res Int, 2014:601568.
6	Zeisberg M, Duffield JS. Resolved: EMT produces fibroblasts in the kidney[J]. J Am Soc Nephrol, 2010, 21(8):1247-1253.
7	He J, Xu Y, Koya D, et al. Role of the endothelial-to-mesenchymal transition in renal fibrosis of chronic kidney disease[J]. Clin Exp Nephrol, 2013, 17(4):488-497.
8	Boutet A, De Frutos CA, Maxwell PH, et al. Snail activation disrupts tissue homeostasis and induces fibrosis in the adult kidney[J]. EMBO J, 2006, 25(23):5603-5613.
9	Boutet A, Esteban MA, Maxwell PH, et al. Reactivation of snail genes in renal fibrosis and carcinomas: a process of reversed embryogenesis?[J]. Cell Cycle, 2007, 6(6):638-642.
10	Camussi G, Deregibus MC, Bruno S, et al. Exosomes/microvesicles as a mechanism of cell-to-cell communication[J]. Kidney Int, 2010, 78(9):838-848.
11	Camussi G, Deregibus MC, Cantaluppi V. Role of stem-cell-derived microvesicles in the paracrine action of stem cells[J]. Biochem Soc Trans, 2013, 41(1):283-287.
12	Zhu YG, Feng XM, Abbott J, et al. Human mesenchymal stem cell microvesicles for treatment of Escherichia coli endotoxin-induced acute lung injury in mice[J]. Stem Cells, 2014, 32(1):116-125.
13	Arslan F, Lai RC, Smeets MB, et al. Mesenchymal stem cell-derived exosomes increase ATP levels, decrease oxidative stress and activate PI3K/Akt pathway to enhance myocardial viability and prevent adverse remodeling after myocardial ischemia/reperfusion injury[J]. Stem Cell Res, 2013, 10(3):301-312.
14	Du T, Zou X, Cheng J, et al. Human wharton's jelly-derived mesenchymal stromal cells reduce renal fibrosis through induction of native and foreign hepatocyte growth factor synthesis in injured tubular epithelial cells[J]. Stem Cell Res Ther, 2013, 4(3):59.
15	Du T, Ju G, Wu S, et al. Microvesicles derived from human Wharton's jelly mesenchymal stem cells promote human renal cancer cell growth and aggressiveness through induction of hepatocyte growth factor[J]. PLoS One, 2014, 9(5):e96836.
16	Li R, Liang J, Ni S, et al. A mesenchymal-to-epithelial transition initiates and is required for the nuclear reprogramming of mouse fibroblasts[J]. Cell Stem Cell, 2010, 7(1):51-63.
17	Majore I, Moretti P, Hass R, et al. Identification of subpopulations in mesenchymal stem cell-like cultures from human umbilical cord[J]. Cell Commun Signal, 2009, 7:6.
18	Zou X, Zhang G, Cheng Z, et al. Microvesicles derived from human Wharton's Jelly mesenchymal stromal cells ameliorate renal ischemia-reperfusion injury in rats by suppressing CX3CL1[J]. Stem Cell Res Ther, 2014, 5(2):40.
19	Devarajan P. Update on mechanisms of ischemic acute kidney injury[J]. J Am Soc Nephrol, 2006, 17(6):1503-1520.
20	Caplan AI, Dennis JE. Mesenchymal stem cells as trophic mediators[J]. J Cell Biochem, 2006, 98(5):1076-1084.
21	Du T, Cheng J, Zhong L, et al. The alleviation of acute and chronic kidney injury by human Wharton's jelly-derived mesenchymal stromal cells triggered by ischemia-reperfusion injury via an endocrine mechanism[J]. Cytotherapy, 2012, 14(10):1215-1227.
22	Zhang G, Zou X, Miao S, et al. The anti-oxidative role of micro-vesicles derived from human Wharton-Jelly mesenchymal stromal cells through NOX2/gp91(phox) suppression in alleviating renal ischemia-reperfusion injury in rats[J]. PLoS One, 2014, 9(3):e92129.
23	Gu D, Zou X, Ju G, et al. Mesenchymal Stromal Cells Derived Extracellular Vesicles Ameliorate Acute Renal Ischemia Reperfusion Injury by Inhibition of Mitochondrial Fission through miR-30[J]. Stem Cells Int, 2016:2093940.
24	Li R, Liang J, Ni S, et al. A mesenchymal-to-epithelial transition initiates and is required for the nuclear reprogramming of mouse fibroblasts[J]. Cell Stem Cell, 2010, 7(1):51-63.
25	Lamouille S, Xu J, Derynck R. Molecular mechanisms of epithelial-mesenchymal transition[J]. Nat Rev Mol Cell Biol, 2014, 15(3):178-196.

分组	样数	TUNEL染色	BrdU染色
空白组	6	10±2	0 ~ 1
EVs组	6	2±1^a	4±1^a
Oct-4过表达组	6	0 ~1^ab	7±2^ab
Oct-4低敲组	6	7±2^abc	2±1^abc
F值	?	50.20	30.57
P值	?	< 0.01	< 0.01

分组	样数	TUNEL染色	BrdU染色
空白组	6	10±2	0 ~ 1
EVs组	6	2±1^a	4±1^a
Oct-4过表达组	6	0 ~1^ab	7±2^ab
Oct-4低敲组	6	7±2^abc	2±1^abc
F值	?	50.20	30.57
P值	?	< 0.01	< 0.01

分组	样数	TUNEL染色		PCNA染色
分组	样数	48 h	2周	48 h	2周
空白组	6	224±25	145±20	5± 2	2±1
EVs组	6	114±23^a	88±12^a	51± 6^a	10±2^a
Oct-4过表达组	6	2± 1^ab	3± 2^ab	70± 7^ab	21±4^ab
Oct-4低敲组	6	134±12^ac	107±14^abc	29±10^abc	5±2^bc
F值	?	153.74	116.29	99.93	66.88
P值	?	< 0.01	< 0.01	< 0.01	< 0.01

分组	样数	TUNEL染色		PCNA染色
分组	样数	48 h	2周	48 h	2周
空白组	6	224±25	145±20	5± 2	2±1
EVs组	6	114±23^a	88±12^a	51± 6^a	10±2^a
Oct-4过表达组	6	2± 1^ab	3± 2^ab	70± 7^ab	21±4^ab
Oct-4低敲组	6	134±12^ac	107±14^abc	29±10^abc	5±2^bc
F值	?	153.74	116.29	99.93	66.88
P值	?	< 0.01	< 0.01	< 0.01	< 0.01

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