Upload
jinyuan
View
224
Download
0
Embed Size (px)
Citation preview
ORIGINAL RESEARCH PAPER
Human adipose-derived stem cells modified by HIF-1aaccelerate the recovery of cisplatin-induced acute renalinjury in vitro
WeiWei Wang • Wei Wang • Yan Jiang •
Zezheng Li • Jin Cheng • Nanmei Liu •
GuoFeng Han • Shi Lu • JinYuan Zhang
Received: 6 April 2013 / Accepted: 8 October 2013
� Springer Science+Business Media Dordrecht 2013
Abstract Human adipose-derived stem cells (hAS-
Cs) improve renal function in acute kidney injury.
Hypoxia-inducible factor-1a (HIF-1a) was transfected
into hASCs. hASCs modified by lentivirus-mediated
empty-vector and HIF-1a maintained their stem cell
characteristics. The expression of the renal-protective
gene, heme oxygenase-1 and vascular endothelial
growth factor were significantly increased in hASCs
modified by HIF-1a, compared to hASCs modified by
empty-vector. Cellular ultra-structure and TUNEL
staining revealed that hASCs modified by HIF-1apromoted the recovery of apoptotic morphology in
cisplatin-treated human kidney-2 cells (HK-2 cells)
when compared to hASCs modified by empty-vector.
Additionally, hASCs modified by empty-vector inhib-
ited caspase-3 expression and up-regulated Bcl-2
expression in cisplatin-treated HK-2 cells, an effect
even more pronounced with hASCs modified by HIF-
1a. Thus, HIF-1a gene-modified ASCs could be an
effective way to enhance the renal-protective effect.
Keywords Acute kidney injury � Cisplatin �Human adipose-derived stem cells � Human
kidney-2 cells � Hypoxia inducible factor-1a �Renal injury
Introduction
Therapeutic options for acute kidney injury (AKI) are
limited to the use of supportive measures and dialysis.
A recent approach that has sparked great interest and
Electronic supplementary material The online version ofthis article (doi:10.1007/s10529-013-1389-x) contains supple-mentary material, which is available to authorized users.
W. Wang � J. Cheng � N. Liu � G. Han �S. Lu � J. Zhang (&)
Department of Nephrology, Jimin Hospital,
Shanghai 200052, People’s Republic of China
e-mail: [email protected]
W. Wang
e-mail: [email protected]
J. Cheng
e-mail: [email protected]
N. Liu
e-mail: [email protected]
G. Han
e-mail: [email protected]
S. Lu
e-mail: [email protected]
W. Wang � Y. Jiang � Z. Li
Graduate School, Shanghai University of Traditional
Chinese Medicine, Shanghai 201203, People’s Republic
of China
e-mail: [email protected]
Y. Jiang
e-mail: [email protected]
Z. Li
e-mail: [email protected]
123
Biotechnol Lett
DOI 10.1007/s10529-013-1389-x
gained popularity is the utilization of stem cells to
repair acutely damaged kidneys. Adipose tissue is an
attractive source of multipotent stem cells that can
differentiate into osteogenic, chondrogenic, myo-
genic, neurogenic, hematopoietic or endothelial cells
(Colazzo et al. 2010; Witkowska-Zimny and Walenko
2011). Additionally, ASCs show a high proliferation
rate and low senescence rate even when harvested
from adults, and they do not trigger immune rejection
(Xishan et al. 2013). Thus, adipose tissue is a
promising tissue source for stem cells with powerful
implications for regenerative medicine.
Hypoxia-inducible factor-1a (HIF-1a), one of HIF
family members, is a master regulator that mediates
the adaptive response to hypoxia in cells and tissues.
HIF-1a plays a key protective role against renal
damage, inducing the expression of downstream renal-
protective genes, including heme oxygenase-1 (HO-1)
and vascular endothelial growth factor (VEGF) (Wang
and Zhang 2008; Weidemann et al. 2008).
Under normoxic conditions, HIF-a is rapidly
degraded because it contains an O2-dependent degra-
dation (ODD) domain. Huang et al. (1998) reported that
cells transfected with cDNA encoding HIF-1a in which
the ODD was deleted showed constitutively active HIF-
1a signaling regardless of its O2 tension and that
deletion of the ODD domain [to generate HIF-1a(DODD)] did not affect the function of HIF-1a (Rosen-
berger et al. 2003), a finding which may be beneficial in
exploring the effect of HIF-1a on damaged tissue under
normoxic conditions. Based on these findings, we
investigated the enhanced protective effect of lentivi-
rus-mediated HIF-1a (DODD) over-expression in hAS-
Cs against cisplatin-induced nephrotoxicity in vitro.
Materials and methods
Human adipose-derived stem cells (hASC) culture
and preparation
hASCs, purchased from Cyagen Biosciences Inc.,
were maintained in culture medium including
Dulbecco’s Modified Eagle Medium (DMEM), 10 %
(v/v) fetal bovine serum, 50 U penicillin ml-1 and
50 lg streptomycin ml-1 at 37 �C/5 % CO2. At 80 %
confluence, cells were trypsinized with 0.25 % tryp-
sin/EDTA and passaged into new flasks for further
expansion. The medium was changed every other day.
Lentivirus production
HIF-1a (DODD) was amplified from pcDNA3-HIF-
1a (401D603; a gift from Dr. Franklin) by PCR using
primers containing BamHI and AscI restriction sites.
HIF-1a (DODD) was constructed into lentivirus-
expressing vector containing green fluorescent protein
(GFP) according to the Invitrogen protocol. Viral
particles were harvested and stored at -80 �C. The
empty lentiviral vector was generated using the same
procedure. (HIF-1a over-expression refers to HIF-1a(DODD) over-expression.)
Lentiviral transfection of hASCs
hASCs were plated in 25 cm2 flasks and grown to
80 % confluence (*106 cells). Cells were incubated
overnight with lentivirus at a multiplicity of infection
(moi) of 1 in the presence of 8 lg polybrene ml-1
(Sigma, USA), and the medium was replaced with
5 ml fresh medium the next day. Three days later,
GFP-expressing cells were collected by FACS and re-
plated for further culture. GFP expression of sorted
cells was examined by fluorescence microscopy.
Additionally, immunohistochemistry and western blot
were used to assess the expression of HIF-1a in hASCs
transduced with empty vector and HIF-1a.
Character of hASCs modified by lentivirus-
mediated empty vector and HIF-1a
hASCs modified by lentivirus-mediated empty vector
(EV-hASCs) and HIF-1a (HIF-1a-hASCs) (2 9 106
cells) were trypsinized, washed three times with PBS,
and immunostained for 30 min on ice with monoclo-
nal antibodies against CD29 (FITC-conjugated),
CD44 (FITC-conjugated) and CD105 (phycoery-
thrin-conjugated). Labeled cells were analyzed using
a flow cytometer. Differentiation potential of EV-
hASCs and HIF-1a-hASCs were examined for oste-
ogenic and adipogenic differentiation according to the
manufacture’s protocol (Cyagen).
Cisplatin-induced AKI model in vitro
and treatments
Human kidney-2 (HK-2 cells, ATCC, USA) induced by
cisplatin, was used as an in vitro AKI model. Cisplatin
(Sigma, 1, 2, 3, and 4 lg ml-1) was used to treat HK-2
Biotechnol Lett
123
cells. 3 lg cisplatin ml-1 was the optimal concentration
for inducing apoptosis in HK-2 cells according to
transmission electron microscopy. We designed exper-
iments in which HK-2 cells were cultured in four
different conditions: untreated cells cultured alone
(control); cisplatin-treated cells cultured alone (cisplatin
3 lg ml-1); cisplatin-treated HK-2 cells cultured with
EV-hASCs; and cisplatin-induced HK-2 cells co-cul-
tured with HIF-1a-hASCs. The detailed protocol was as
follows. HK-2 cells were seeded at 2 9 103 cells/cm2 in
six-well plates in keratinocyte serum-free medium
(Invitrogen, USA) supplemented with 2 % (v/v) fetal
bovine serum and incubated for 3 days. Cells were
starved for 12 h to induce synchronization. Control cells
were then cultured in DMEM/F12 supplemented with
5 % (v/v) fetal bovine serum for 6 h, and the three
experimental groups were cultured with DMEM/F12,
5 % (v/v) fetal bovine serum plus cisplatin (3 lg ml-1)
for 6 h. After 6 h, all cells were washed three times with
DMEM. Control and cisplatin-treated cells were then
cultured in DMEM/F12 and 5 % (v/v) fetal bovine
serum for 24 h, and the two co-culture groups were
cultured with EV-hASCs or HIF-1a-hASCs using the
six-well plate transwell co-culture system (Corning,
USA) according to the manufacturer’s instruction.
Briefly, EV-hASCs and HIF-1a-hASCs were first
seeded on polycarbonate inserts (105 cells per insert)
and cultured for 24 h. Inserts were then transferred into
six-well plates containing cisplatin-pretreated HK-2
cells, and cells were cultured together in DMEM/F12
and 5 % (v/v) fetal bovine serum for 24 h. After 24 h,
HK-2 cells were harvested for the following analyses.
Transmission electron microscopy
HK-2 cells were fixed with 2 % (w/v) glutaraldehyde,
dehydrated and embedded in epoxy resin. Ultrathin
sections of 70 nm were prepared and stained with lead
citrate. The ultra-structure of HK-2 cells was observed
using transmission electron microscopy.
Terminal deoxynucleotidyltransferase-mediated
dUTP nick end-labelling (TUNEL) staining
TUNEL staining was performed using the in situ
Apoptosis Detection Kit (Boster Biotech Co, China)
according to the manufacturer’s instruction. Briefly,
HK-2 cells were fixed, endogenous peroxidase was
quenched, and the cells were permeabilised. The cells
were then incubated with the TUNEL reaction
mixture, 3,30-diaminobenzidine tetrahydrochloride
(DAB), and haematein. The apoptotic index was
calculated by the percentage of positive nuclei.
Immunocytochemistry
Immunocytochemical staining was performed using
the SABC system (Boster Biotech Co, China) accord-
ing to the manufacturer’s instruction. Briefly, EV-
hASCs or HIF-1a-hASCs was seeded at 2 9 104 cells/
cm2 on coverslips coated with poly-L-lysine. After
5 days, cells were fixed with acetone for 30 min at
4 �C and washed with PBS three times. Coverslips
were incubated with HIF-1a monoclonal antibody
(Abcam, USA; 1:1,000) overnight at 4 �C followed by
incubation with a suitable secondary antibody. Chro-
mogenic detection was achieved with DAB.
Western blotting
HK-2 cells were rinsed twice with cold PBS and lysed
in whole-cell lysis buffer containing a protease
inhibitor cocktail. Equal amounts of protein were
separated by SDS-PAGE and transferred onto a
nitrocellulose membrane. The membranes were
blocked with 5 % (v/v) nonfat milk in TBS-T
(10 mM Tris-buffered saline with 0.1 % Tween 20)
and incubated with HIF-1a monoclonal antibody
(MAB1536, R&D, USA) overnight at 4 �C. The
membranes were then incubated with the appropriate
secondary antibodies conjugated to horseradish per-
oxidase (HRP; Sigma) (at 1:5,000 dilution) and the
proteins were visualized using enhanced chemilumi-
nescence (ECL) and exposure to X-ray film (Kodak).
Enzyme-linked immunosorbent assay
Culture supernatants from control, model, EV-hASCs
and HIF-1a-hASCs were collected and the level of
HO-1 and VEGF in the medium was determined
according to the manufacturer’s instruction (R&D,
USA).
Analysis of mRNA expression by real time-PCR
Total RNA was isolated from the cells using TRIZOL
and the reverse transcription of the purified RNA was
performed using oligo (dT) priming and superscript II
Biotechnol Lett
123
reverse transcription, according to the manufacturer’s
instruction (Invitrogen, USA). Real time-PCR was
performed using SYBR green. The primer pairs for the
selected genes are listed in Supplementary Table 1.
Statistical analysis
Samples values are expressed as the mean ± standard
deviation (SD). Data were analyzed by ANOVA using
the SPSS13.0 statistical software package. P-values
less than 0.05 were considered significant.
Results
Efficient transduction of hASCs by lentivirus
mediated-empty vector and HIF-1a
After transient transduction with lentivirus, EV-hAS-
Cs or HIF-1a-hASCs were cultured for 72 h, and then
were observed by phase contrast and fluorescence
microscopy (Fig. 1Aa, b, e, f). After FACS sorting of
GFP-expressing hASCs, we achieved [99 % GFP
positivity as assessed by microscopy at 48 h (Fig. 1Ac,
d, g, h). Expression of HIF-1a was markedly increased
in HIF-1a-hASCs, as demonstrated by western blotting
(Fig. 1B) and immunocytology (Fig. 1C).
Differentiation and surface markers of hASCs
modified by lentivirus-mediated empty vector
and HIF-1a
To evaluate multi-differentiation of EV-hASCs and
HIF-1a-hASCs, both types of hASCs were differentially
induced into osteoblasts and adipocytes, respectively.
After 3 weeks’ incubation, both types of hASCs became
Alizarin Red positive with osteogenic supplementation
(Fig. 2Aa, b) and, were induced with adipogenic
medium, showed Oil Red O positive lipid droplets
(Fig. 2Ac, d). Flow cytometry analysis showed that EV-
hASCs and HIF-1a-hASCs were positive for mesen-
chymal markers CD29, CD44 and CD105 (Fig. 2B).
Evaluation of HO-1 and VEGF expression
To measure the protein expression of HO-1 and VEGF
in the different medium, ELISA was used. HO-1 and
VEGF protein expression in the medium of HK-2 cells
incubated with 3 lg cisplatin ml-1 were significantly
decreased compared to that in the medium of control
cells (P \ 0.05). After co-culture of the cisplatin-
treated HK-2 cells with EV-hASCs or HIF-1a-hASCs,
HO-1 and VEGF protein expression were increased,
and there was a significant difference between both co-
cultured groups and cisplatin-induced cells (P \ 0.05).
Significant differences were also observed between the
cells co-cultured with EV-hASCs and HIF-1a-hASCs
(P \ 0.05) (Fig. 3a and b). Additionally, HO-1 and
VEGF gene expression were also evaluated by real-
time PCR, with the results showing the same trend as
the protein expression (Fig. 3c and d).
Fig. 1 Virus transduction of hASCs. A Transiently transfected
hASCs (a, b and e, f) and stably transfected hASCs (c, d and g, h) in
the EV-hASC and HIF-1a-hASC groups were sorted by FACScan
after 72 h. Western blot (B) and immunocytochemistry (C) anal-
yses were performed for HIF-1a protein expression in the EV-
hASC and HIF-1a-hASC groups (n = 3 each). Scale bar 100 lm
Biotechnol Lett
123
Fig. 2 Characteristics of
EV-hASCs and HIF-1a-
hASCs. A Multi-potential
differentiation of EV-
hASCs and HIF-1a-hASCs,
including osteoblasts
(Alizarin Red, a, b) and
adipocytes (Oil Red O, c, d).
Scale bar 100 lm. B Flow
cytometric analysis showing
EV-hASC and HIF-1a-
hASC positivity for
mesenchymal markers
CD29, CD44, and CD105
(n = 3 each)
Biotechnol Lett
123
Ultra-structural analysis of HK-2 cells
To determine the optimal cisplatin concentration for
inducing cellular apoptosis in HK-2 cells, transmission
electron microscopy was used to observe the change in
cellular structure. Untreated control HK-2 cells
appeared normal (Fig. 4a). After treatment with 1 lg
cisplatin ml-1 for 6 h, HK-2 cells did not show obvious
alterations in cellular structure (Fig. 4b). HK-2 cells
treated with 2 lg cisplatin ml-1 presented cellular
chromatin condensation (Fig. 4c), and those treated
with 3 lg cisplatin ml-1 showed typical apoptotic
characteristics (Fig. 4d). HK-2 cells treated with 4 lg
cisplatin ml-1 underwent necrosis (Fig. 4e). When HK-
2 cells exposed to 3 lg cisplatin ml-1 were co-cultured
with EV-hASCs, those cells appeared less apoptotic
(Fig. 4f). When HK-2 cells treated with 3 lg cisplatin
ml-1 were co-cultured with with HIF-1a-hASCs, most
cells recovered their normal cellular structure, with only
a few cells exhibiting modest cellular damage, such as
swollen mitochondria or heterochromatic foci (a char-
acteristic of early apoptosis) (Fig. 4g).
Fig. 3 The level of HO-1 and VEGF protein and gene expression. The data are expressed as the mean ± SD (n = 5 each) *P \ 0.05,
compared to the model group. #P \ 0.05, compared to the EV-hASCs group
Biotechnol Lett
123
DNA fragmentation determined by TUNEL assay
To detect DNA fragmentation in situ and calculate the
apoptotic index, TUNEL staining was performed.
Treatment with 3 lg cisplatin ml-1 increased the
apoptotic index in the model and a significant
difference was observed between the model and
control groups (P \ 0.05). However, co-culturing
HK-2 cells with EV-hASCs and HIF-1a-hASCs
lowered the apoptotic index, with a significant
Fig. 4 Ultra-structural change of HK-2 cells. a Control
(untreated cells cultured alone); b incubated with 1 lg cisplatin
ml-1; c incubated with 2 lg cisplatin ml-1; d incubated with
3 lg cisplatin ml-1; e incubated with 4 lg cisplatin ml-1;
f cisplatin (3 lg ml-1)-induced cells co-cultured with EV-
hASCs; g cisplatin (3 lg ml-1)-induced cells co-cultured with
HIF-1a-hASCs. The arrows show cellular alterations (a, normal
cellular shape; b, no obvious changes; c, chromatin condensa-
tion; d, typical apoptotic characteristics; e, necrosis; d and f, the
recovery of cellular structure). Scale bar (a, b, c, g): 10 lm,
Scale bar (d, e, f), 3 lm
Biotechnol Lett
123
difference between the co-incubated groups and
model group (P \ 0.05) (Fig. 5).
Analysis of caspase-3 and Bcl-2 gene expression
To evaluate the expression of related apoptotic
gene, real time-PCR was used. Cisplatin (3 lg ml-1)
increased the gene expression of caspase-3 and
decreased the gene expression of Bcl-2 in HK-2 cells
compared to control cells (P \ 0.05). Co-culturing
HK-2 cells with EV-hASCs and HIF-1a-hASCs
resulted in decreased levels of caspase-3 expression
and prevented the cisplatin-induced reduction in Bcl-2
gene expression in HK-2 cells. The EV-hASCs and
Fig. 5 Apoptotic analysis.
A TUNEL staining.
a. control (untreated cells
cultured alone); b. incubated
with cisplatin (3 lg ml-1);
c. cisplatin (3 lg ml-1)-
induced cells co-cultured
with h EV-hASCs;
d. cisplatin (3 lg ml-1)-
induced cells co-cultured
with HIF-1a-hASCs. Scale
bar 100 lm. B Apoptotic
index. The data are expressed
as the mean ± SD (n = 5
each) *P \ 0.05, compared
to the model group
Biotechnol Lett
123
HIF-1a-hASCs groups showed significant differences
compared to the model group (P \ 0.05) (Fig. 6).
Discussion
We have delivered human HIF-1a gene into hASCs
via a lentiviral vector and increased the levels of HIF-
1a expression in vitro. EV-hASCs or HIF-1a-hASCs
maintained their stem cell characteristics, including
the expression of surface antigens and normal differ-
entiation potential. Additionally, EV-hASCs secreted
some renal-protective genes, HO-1 and VEGF, and
HIF-1a-hASCs was obviously promoted the expres-
sion of HO-1 and VEGF.
The main forms of cisplatin-induced renal cell
injury are apoptosis and necrosis and the pathway of
cell death is concentration dependent, with low
concentrations of cisplatin primarily inducing apop-
tosis and high concentrations predominantly inducing
necrosis. In this study, 3 lg cisplatin ml-1 was
regarded as optimal for inducing apoptosis in HK-2
cells. Unlike necrosis, apoptosis is mediated by the
active participation of the dying cells. Therefore,
apoptosis appears much easier to reversal by some
methods compared with unreversible necrosis.
Stem cells, especially mesenchymal stem cells
(MSCs), have protective effects against AKI arising
from chemical (glycerol and cisplatin) and ischemia–
reperfusion injury by secreting beneficial factors and
activating signal proteins (Kim et al. 2012; Zarjou and
Agarwal 2012). Conversely, beneficial effects were
not reported for CsA-induced renal injury (Chung
et al. 2013). Our study demonstrates that EV-hASCs
may improve the cellular morphology of co-cultured
HK-2 cells incubated with cisplatin suggesting that
EV-hASCs exert their renal protective function
through an autocrine mechanism, such as HO-1 and
VEGF expression. Zarjou et al. (2011) study indicated
that the conditioned medium of HO-1?/?MSCs
rescued the functional and morphological changes
associated with cisplatin-induced AKI, whereas an
HO-1-/--conditioned medium was ineffective. In
addition, Togel et al. (2009) study showed that VEGF
is an important mediator of the early and late phase of
renoprotective action after AKI within the context of
stem cell treatment. We also explored the mechanism
for the EV-hASC-mediated protection of cisplatin-
treated HK-2 cells. Caspase-3 is activated by mito-
chondrial injury leading to an increase in cytochrome c
release into the cytoplasm, a process that was precisely
regulated by members of the Bcl-2 family (Zheng et al.
2013). In our study, EV-hASCs suppressed the
cisplatin-induced activation of caspase-3 suggesting
that EV-hASCs may act upstream of caspase-3 to
block apoptosis. Based on our observations, a decrease
in caspase-3 expression correlated well with an
increase in anti-apoptotic Bcl-2 expression.
Genetically-modified stem cells over-expressing
particular genes can have an enhanced protective
effect on damaged cells and tissues, perhaps through a
paracrine mechanism (Lu et al. 2008; Tadagavadi and
Fig. 6 Analysis of caspase-3 and BcL-2 gene expression. The
data are expressed as the mean ± SD (n = 5 each) *P \ 0.05,
compared to the model group. #P \ 0.05, compared to the EV-
hASCs group
Biotechnol Lett
123
Reeves 2010). In our study, HIF-1a-hASCs improved
the cellular morphology of cisplatin-treated HK-2
cells to a greater degree than the parental hASCs.
Elsewhere, activation of HIF by DMOG halted the
progression of proteinuria, attenuated structural dam-
age and decreased oxidative stress, inflammation, and
fibrosis in a remnant kidney model (Zarjou et al. 2011)
and TRC160334, a novel HIF hydroxylase inhibitor,
stabilized HIF-a and activated HIF, a situation that led
to a significant reduction in renal injury and serum
creatinine and the improvement of urine output in AKI
(Jamadarkhana et al. 2012). Additionally, activation of
HIF by cobalt or DMOG attenuated renal dysfunction,
proteinuria, and structural damage through a reduction
of oxidative stress, inflammation, and apoptosis in
renal tubular epithelial cells in gentamicin-induced
AKI (Ahn et al. 2012). The mechanisms through
which hASCs protected HK-2 cells from cisplatin-
induced cell apoptosis included promoting the secre-
tion of HO-1 and VEGF, decreasing activation of
caspase-3 and increasing expression of Bcl-2, which
showed an effect even more pronounced with hASCs
modified by HIF-1a.
In conclusion, our study indicated that hASCs and
HIF-1a-modified hASCs show a protective effect on
cisplatin-induced AKI in vitro and HIF-1a-modified
hASC was more efficacious in treating AKI in vitro.
Acknowledgments This research was supported by National
Natural Science Foundation of China (No. 81100493), Key
Project of Basic Research of Science and Technology of Shanghai
(12DJ1400203), Shanghai Rising-Star Program (09QA1407500)
and Project of Shanghai Excellent Young Doctor (XYQ2011012).
References
Ahn JM, You SJ, Lee YM, Oh SW, Ahn SY, Kim S, Chin HJ,
Chae DW, Na KY (2012) Hypoxia-inducible factor acti-
vation protects the kidney from gentamicin-induced acute
injury. PLoS One 7:e48952
Chung BH, Lim SW, Doh KC, Piao SG, Heo SB, Yang CW
(2013) Human adipose tissue derived mesenchymal stem
cells aggravate chronic cyclosporin nephrotoxicity by the
induction of oxidative stress. PLoS One 8:e59693
Colazzo F, Chester AH, Taylor PM, Yacoub MH (2010)
Induction of mesenchymal to endothelial transformation of
adipose-derived stem cells. J Heart Valve Dis 19:736–744
Huang LE, Gu J, Schau M, Bunn HF (1998) Regulation of
hypoxia-inducible factor 1alpha is mediated by an O2-
dependent degradation domain via the ubiquitin-protea-
some pathway. Proc Natl Acad Sci USA 95:7987–7992
Jamadarkhana P, Chaudhary A, Chhipa L, Dubey A, Mohanan
A, Gupta R, Deshpande S (2012) Treatment with a novel
hypoxia-inducible factor hydroxylase inhibitor (TRC160334)
ameliorates ischemic acute kidney injury. Am J Nephrol
36:208–218
Kim JH, Park DJ, Yun JC, Jung MH, Yeo HD, Kim HJ, Kim
DW, Yang JI, Lee GW, Jeong SH, Roh GS, Chang SH
(2012) Human adipose tissue-derived mesenchymal stem
cells protect kidneys from cisplatin nephrotoxicity in rats.
Am J Physiol Renal Physiol 302:F1141–F1150
Lu LH, Oh DJ, Dursun B, He Z, Hoke TS, Faubel S, Edelstein
CL (2008) Increased macrophage infiltration and fractal-
kine expression in cisplatin-induced acute renal failure in
mice. J Pharmacol Exp Ther 324:111–117
Rosenberger C, Griethe W, Gruber G, Wiesener M, Frei U,
Bachmann S, Eckardt KU (2003) Cellular responses to
hypoxia after renal segmental infarction. Kidney Int 64:
874–886
Tadagavadi RK, Reeves WB (2010) Endogenous IL-10 atten-
uates cisplatin nephrotoxicity: role of dendritic cells.
J Immunol 185:4904–4911
Togel F, Zhang P, Hu Z, Westenfelder C (2009) VEGF is a
mediator of the renoprotective effects of multipotent
marrow stromal cells in acute kidney injury. J Cell Mol
Med 13:2109–2114
Wang W, Zhang J (2008) Induction of renoprotective gene
expression by hypoxia-inducible transcription factor-
1alpha ameliorates renal damage. Med Hypotheses 70:
948–950
Weidemann A, Bernhardt WM, Klanke B, Daniel C, Buchholz
B, Campean V, Amann K, Warnecke C, Wiesener MS,
Eckardt KU, Willam C (2008) HIF activation protects from
acute kidney injury. J Am Soc Nephrol 19:486–494
Witkowska-Zimny M, Walenko K (2011) Stem cells from adi-
pose tissue. Cell Mol Biol Lett 16:236–257
Xishan Z, Baoxin H, Xinna Z, Jun R (2013) Comparison of the
effects of human adipose and bone marrow mesenchymal
stem cells on T lymphocytes. Cell Biol Int 37:11–18
Zarjou A, Agarwal A (2012) Heme oxygenase-1 as a target for
TGF-b in kidney disease. Semin Nephrol 32:277–286
Zarjou A, Kim J, Traylor AM, Sanders PW, Balla J, Agarwal A,
Curtis LM (2011) Paracrine effects of mesenchymal stem
cells in cisplatin-induced renal injury require heme oxy-
genase-1. Am J Physiol Renal Physiol 300:F254–F262
Zheng Y, Lu M, Ma L, Zhang S, Qiu M, Wang Y (2013) Osthole
ameliorates renal ischemia-reperfusion injury in rats.
J Surg Res 183:347–354
Biotechnol Lett
123