Spirulina attenuates cyclosporine-induced nephrotoxicity in rats
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Transcript of Spirulina attenuates cyclosporine-induced nephrotoxicity in rats
444 M. KHAN ET AL.
Copyright © 2006 John Wiley & Sons, Ltd. J. Appl. Toxicol. 2006; 26: 444–451
DOI: 10.1002/jat
JOURNAL OF APPLIED TOXICOLOGYJ. Appl. Toxicol. 2006; 26: 444 –451Published online 20 July 2006 in Wiley InterScience(www.interscience.wiley.com) DOI: 10.1002/jat.1159
Spirulina attenuates cyclosporine-induced nephrotoxicityin rats
Mahmood Khan,1 Jagdish Chandra Shobha,1 Iyyapu Krishna Mohan,1 Madireddy Umamaheswara RaoNaidu,1 Aruna Prayag2 and Vijay Kumar Kutala1*
1 Department of Clinical Pharmacology and Therapeutics, Nizam’s Institute of Medical Sciences, Punjagutta, Hyderabad 500 082,India
2 Department of Pathology, Nizam’s Institute of Medical Sciences, Punjagutta, Hyderabad 500 082, India
Received 22 December 2005; Revised 7 March 2006; Accepted 12 May 2006
ABSTRACT: Cyclosporine (CsA) causes a dose-related decrease in renal function in experimental animals and humans.
The generation of reactive oxygen species (ROS) has been implicated in CsA-induced nephrotoxicity. It was previously
shown that Spirulina, a blue-green algae, with antioxidant properties effectively attenuated the doxorubicin-induced
cardiotoxicity in mice and cisplatin-induced nephrotoxicity in rat. The present study investigated the nephroprotective role
of Spirulina against CsA-induced nephrotoxicity in rats. Spirulina (500 mg kg−−−−−1 b.w.) was administered orally for 3 days
before and 14 days concurrently with CsA (50 mg kg−−−−−1 b.w.). Rats treated with CsA showed nephrotoxicity as evidenced
from a significant elevation in plasma urea, creatinine, urinary N-acetyl-βββββ -D-glucosaminidase (βββββ -NAG) and a decrease in
creatinine and lithium clearance. Pretreatment with Spirulina protected the rats from CsA-induced nephrotoxicity. The
CsA-induced rise in plasma urea and creatinine and the decrease in creatinine and lithium clearance were attenuated by
Spirulina. There was a significant increase in plasma and kidney tissue MDA with CsA. Spirulina prevented the rise in
plasma and kidney tissue MDA. Histopathology of the kidney from CsA-treated rats showed severe isometric vacuolization
and widening of the interstitium. However, pretreatment with Spirulina prevented such changes, and the kidney morpho-
logy was comparable to that of the control. Spirulina treatment did not alter the blood CsA levels. These results suggest
that Spirulina has a protective effect against nephrotoxicity induced by CsA. This study further supports the crucial role
of the antioxidant nature of Spirulina in protecting against CsA-induced oxidative stress. Copyright © 2006 John Wiley
& Sons, Ltd.
KEY WORDS: cyclosporine; nephrotoxicity; Spirulina; oxidative stress; antioxidant
increased synthesis of endothelin (Fogo et al., 1992;
Lanese and Conger, 1993), induction of cytochrome P450
enzymes in renal microsomes (Serino et al., 1994) and
oxidative stress (De Nicola et al., 1993). Recent studies
have also suggested that nitric oxide is involved in the
haemodynamic alterations encountered with CsA treat-
ment by modulating the activity of the inducible form of
nitric oxide synthase (iNOS) in renal tissues (Amore et
al., 1995; Dusting et al., 1999).
The role of exogenous antioxidants on the renal effects
of CsA have been studied extensively (Durak et al., 1998;
Parra et al., 1998; Kumar et al., 1999; L’Azou et al.,
1999). Studies showed that N-acetylcysteine (Tariq et al.,
1999) and α-tocopherol (Wolf et al., 1994) reversed the
CsA-induced nephrotoxicity. The protection of allopurinol
against CsA-induced nephrotoxicity is related to the
reduced formation of oxygen free radicals, preventing
the deleterious effects of lipid peroxidation (Assis et al.,
1997). The effect of lacidipine, a new calcium channel
blocker with antioxidant property has been shown to
protect against CsA-induced nephrotoxicity in rats
(Naidu et al., 1999). It has been hypothesized that the
Introduction
Cyclosporine (CsA) has improved the patient and the
graft survival rates in solid organ transplantation. How-
ever, the therapeutic benefits of CsA are often limited
by the occurrence of acute and chronic nephrotoxicity
which continues to be a major problem (Andoh et al.,
1997). About 30% of the patients treated with CsA
have moderate to severe renal toxicity. CsA causes
unique functional and structural nephrotoxicity. CsA
administration leads to the loss of proximal tubular
epithelial integrity and tubular atrophy, vacuolization and
microcalcification (Rooth et al., 1987). Several mecha-
nisms have been proposed in CsA-induced nephrotoxicity
namely: the activation of the renin–angiotensin system
and enhanced sympathetic tone (Murray et al., 1985),
* Correspondence to: Vijay Kumar Kutala, PhD, The Ohio State University,
420 West 12th Ave, TMRF- Room 126, Columbus, OH 43210, USA.
E-mail: [email protected]
SPIRULINA ON CSA-INDUCED NEPHROTOXICITY 445
Copyright © 2006 John Wiley & Sons, Ltd. J. Appl. Toxicol. 2006; 26: 444–451
DOI: 10.1002/jat
CsA-induced increase in the P450 system may result in
increased free radical formation (Serino et al., 1994). Re-
cent studies have clearly demonstrated that CsA-induced
oxidative stress plays a pivotal role in producing struc-
tural and functional impairment of the kidney (Ichikawa
et al., 1994; Kumar et al., 1999; Shifow et al., 2000).
Spirulina (SP), a blue-green algae, contains proteins,
lipids and carbohydrates, as well as elements such as
selenium, zinc, magnesium and vitamins (Ciferri, 1983).
Spirulina is used as a food supplement and the nutritional
and therapeutic values have been well documented (Kay,
1991). Spirulina is also a good source of antioxidants
such as β-carotene and C-phycocyanin. The extracts of
Spirulina have been known to possess significant anti-
oxidant activity both in vitro and in vivo (Miranda et al.,
1998). C-phycocyanin, a bliiprotein, is a potent free
radical scavenger (Bhat and Madyastha, 2000). Recent
studies have suggested that Spirulina significantly
attenuated lead-induced changes in the levels of lipid
peroxidation and endogenous antioxidants in all the
major tissues of rats (Upasani and Balaraman, 2003).
C-phycocyanin was demonstrated to protect rats from
carbon tetrachloride-induced hepatotoxicity (Vadiraja
et al., 1998). Spirulina was reported to reduce the hepatic
cytochrome P450 content and to increase the hepatic
glutathione S-transferase activity and to be involved
in the activation/detoxification of chemical mutagens/
carcinogens (Mittal et al., 1999). A recent study demon-
strated that Spirulina protected against cisplatin-induced
nephrotoxicity in the rat (Mohan et al., 2006) and
doxorubicin-induced cardiotoxicity in mice (Khan et al.,
2005). The aim of the present study was to investigate
the effect of Spirulina extract against CsA-induced
nephrotoxicity in rats using biochemical and histomor-
phological parameters indicative of nephrotoxicity and
oxidative stress. The results indicated that rats pretreat-
ment with Spirulina significantly attenuated the CsA-
induced nephrotoxicity.
Materials and Methods
Spirulina, a fine dark blue-green spray-dried powder
prepared from Spirulina platensis (batch No. 0027)
was obtained from New Ambadi Estates (P) Ltd, Tamil
Nadu, India. The Spirulina used in the present study
contained proteins (65.38%), minerals (7.95%), total
carotenoids (4.33 mg g−1), β-carotene (1.67 mg g−1), crude
phycocyanin (15.4%) and total pheophorbide (0.020%).
Animals and Treatment
Male Wistar rats (weight 200–250 g) were used for the
study. The rats were housed under conditions of control-
led temperature and a 12 h lighting cycle and were fed
with standard rat chow. The animals were divided
into four groups of ten animals each. The first group
received only normal saline daily for 17 days and served
as the control. The second group received only Spirulina
500 mg kg−1 b.w., orally, daily for 17 days. The third
group received CsA 50 mg kg−1 b.w., (Sandoz Pharma
Ltd, Switzerland) orally, daily for 14 days. The fourth
group received pretreatment with Spirulina 500 mg kg−1
b.w., orally for 3 days and then concurrently with
CsA (50 mg kg−1) for 14 days. This study was approved
by the Institutional Ethical Committee for the use of
animals.
Biochemical Assays
On day 14 of CsA administration, the animals were kept
in individual metabolic cages for 24 h urine collection.
Blood was collected by the ocular puncture method in a
heparinized tube. Plasma urea and creatinine were meas-
ured by commercially available kits using an UV-visible
spectrophotometer (Schimadzu, UV-1604). The trough
whole blood CsA levels in CsA and Spirulina plus CsA
treated rats was measured by using a solid phase extrac-
tion column prior to chromatographic separation on a µ-
bond Bondapak C18 column (3.9 × 300 mm i.d., 10 µm
particle sizes, Waters, Milford, USA) by HPLC (Salm
et al., 1993). The urinary β-NAG activity was measured
as per the procedure described (Horak et al., 1981).
Lithium Clearance Studies
Rats in all the treatment groups received lithium chloride
(5 mmol l−1) in freely accessible drinking water, until the
end of the experiment (Whiting and Simpson, 1988).
Studies have shown that such lithium supplementation
has no effect on the glomerular tubular function or renal
structure (Whiting and Simpson, 1988). At the end of day
14 of CsA treatment, plasma and urinary lithium levels
were determined using a flame photometer and then the
lithium clearance was calculated.
Lipid Peroxidation Products
The amount of lipid peroxidation (MDA) in plasma and
kidney tissue was determined by measuring thiobarbituric
acid reactive substances (TBARS) (Bernheim et al.,
1948). Portions of kidney tissue were dissected and
homogenized in phosphate buffered saline (pH 7.4). The
homogenate was then centrifuged and the supernatant
was collected and precipitated with 20% trichloroacetic
acid and centrifuged. To the protein-free supernatant,
0.33% thiobarbituric acid (TBA) was added and boiled
for 1 h at 95 °C; the TBA reactive products were
446 M. KHAN ET AL.
Copyright © 2006 John Wiley & Sons, Ltd. J. Appl. Toxicol. 2006; 26: 444–451
DOI: 10.1002/jat
extracted in butanol and the intensity of the pink colour
was read at 520 nm. Fresh diluted tetramethoxy propane
(Sigma Chemical Co., USA) was used as the standard.
The protein content of the homogenate was measured and
the result was expressed as nmol MDA-equivalents per
milligram protein. A similar procedure was followed to
measure TBARS in plasma, using 0.5 ml plasma instead
of homogenate.
Estimation of Antioxidant Enzymes
Superoxide dismutase (SOD) activity was determined in
kidney tissue homogenate by the cytochrome c reduction
method using a xanthine–xanthine oxidase–superoxide
generating system (McCord and Fridovich, 1969). The
SOD activity was determined from a standard curve of
the percentage inhibition of cytochrome c reduction with
a known SOD activity. Catalase activity was determined
by the method of Aebi (1984), with H2O2 (10 mM) and
phosphate buffer (0.05 M, pH. 7.0) at 210 nm. A unit is
defined as the amount of enzyme that catalysed the
dismutation of 1 µmol of H2O2 min−1 and expressed as
units mg−1 protein. Glutathione peroxidase activity was
measured by the NADPH oxidation method (Paglia and
Valentine, 1967) and expressed as nmol of NADPH oxi-
dized to NADP mg−1 protein. Protein was determined by
the method of Lowry et al. (1951).
Histopathological Studies
Kidney from all the four groups were fixed in 10%
buffered formalin and processed with paraffin wax.
Sections (5 µm) were stained with haematoxylin and
eosin and periodic acid Schiff’s stain (PAS) to detect
calcification. A histomorphological evaluation of all the
kidney sections was carried out in a blinded fashion by a
pathologist who was unaware of the treatment groups.
Statistical Analysis
The statistical significance of differences among values of
individual parameters was evaluated by Student’s t-test.
All the values are expressed as mean ± SD. The signifi-
cance was set at P < 0.05.
Results
Body Weight and Renal Function
Body weight and food intake in all the groups were
comparable at the beginning of the study. There were
no significant differences in the body weight, urinary
flow rate, plasma urea, creatinine, creatinine and lithium
clearance, and urinary β-NAG in the control and
Spirulina (500 mg kg−1) treated groups. In rats treated
with CsA, functional and histological changes in the
kidney were observed. The mean percentage decrease
in the body weight in CsA-treated group was 6.9 from
baseline, and this was attenuated in the Spirulina treated
animals (Table 1). The urinary flow rate (UFR) was
significantly elevated in the CsA treated group (6.05 ±0.14 ml h−1 kg−1) compared with the control (3.63 ±0.22 ml h−1 kg−1) and this was significantly decreased
in the Spirulina + CsA treated group. The urinary β-NAG
activity was significantly increased in the CsA-treated
animals (Table 1). The CsA-induced increase in
urinary β-NAG activity was significantly attenuated by
Spirulina.
Plasma urea and creatinine was significantly in-
creased after CsA treatment. Pretreatment of rats with
Spirulina significantly reduced the CsA-induced in-
crease in plasma urea and creatinine levels (Fig. 1).
Plasma urea levels in the control, Spirulina, CsA
and CsA + Spirulina were 31.2 ± 2.3, 29.8 ± 3.5, 98 ±1.2 and 46.5 ± 5.6 mg dl−1 (Fig. 1a) and creatinine
levels were 0.46 ± 0.02, 0.44 ± 0.03, 1.02 ± 0.15
and 0.44 ± 0.05 mg dl−1, respectively (Fig. 1b). There
was a significant decrease in creatinine clearance (Ccr)
in the CsA (153 ± 26.6 ml min−1) compared with the
control (259 ± 13 ml min−1) groups, whereas treatment
with Spirulina prevented the CsA-induced decrease
in Ccr (223 ± 27 ml min−1) (Fig. 2a). Similarly, in
CsA treated animals, lithium clearance (Licr) was signifi-
cantly decreased compared with the control and this
decrease was prevented by Spirulina pretreatment
(Fig. 2b).
Table 1. Effect of Spirulina on CsA-induced change in body weight and renalparameters
Parameter Control SP CsA CsA + SP
Change in body wt (%) + 4.1 + 4.3 −6.9 −2.8
UFR (ml h−1 kg−1) 3.63 ± 0.22 3.71 ± 0.3 6.05 ± 0.14a 4.29 ± 0.18b
Urinary β -NAG(U l−1) 1.25 ± 0.75 1.31 ± 0.51 2.67 ± 0.82a 1.42 ± 0.67b
SP, Spirulina; CsA, cyclosporine. Values are expressed as mean ± SD (n = 10). a P < 0.05 vs Control; b P < 0.05 vs
CsA.
SPIRULINA ON CSA-INDUCED NEPHROTOXICITY 447
Copyright © 2006 John Wiley & Sons, Ltd. J. Appl. Toxicol. 2006; 26: 444–451
DOI: 10.1002/jat
Figure 1. Effect of Spirulina on (a) plasma urea; (b)plasma creatinine levels in CsA treated rats. Rats weretreated with CsA (50 mg kg−1) and Spirulina(500 mg kg−1) as per the protocol given in Materialsand Methods section. Values are expressed as mean ±SD (n = 10), * P < 0.05 vs control; ** P < 0.05 vs CsA.The results show that Spirulina treatment preventedthe CsA-induced nephrotoxicity
Figure 2. Effect of Spirulina (SP) on (a) creatinineclearance (Ccr), and (b) lithium clearance (LiCr) in CsAtreated rats. Rats were treated with CsA (50 mg kg−1)and Spirulina (500 mg kg−1) as per the protocol given inMaterials and Methods section. Values are expressed asmean ± SD (n = 10), * P < 0.05 vs control; ** P < 0.05 vsCsA. The results show that Spirulina treatment attenu-ated the CsA-induced decrease in glomerular function
Lipid Peroxidation and Antioxidant Enzymes
In CsA treated rats, there was a significant increase
in plasma and renal tissue MDA levels compared with
the control group. Spirulina pretreatment significantly
attenuated the CsA-induced increase in plasma MDA and
kidney tissue MDA (Table 2). The activities of SOD,
catalase and glutathione peroxidase in kidney tissue were
determined in all the groups and the results are shown in
Table 2. In CsA-treated animals the activities of SOD,
catalase and glutathione peroxidase were significantly
decreased compared with the control. Pretreatment with
Spirulina significantly prevented the CsA-induced de-
crease in SOD, catalase and glutathione peroxidase.
Table 2. Effect of Spirulina on CsA-induced changes in MDA and antioxidant enzymes
Parameter Control SP CsA CsA + SP
Plasma MDA(nm) 1.41 ± 0.06 1.43 ± 0.04 2.74 ± 0.58a 1.87 ± 0.19b
Kidney tissue MDA 2.56 ± 0.18 2.48 ± 0.18 4.31 ± 0.58a 2.87 ± 0.51b
(nm mg−1 protein)
SOD (U mg−1 protein) 2.15 ± 0.19 2.19 ± 0.08 1.32 ± 0.43a 2.32 ± 0.20b
Catalase (U mg−1 protein) 0.35 ± 0.02 0.33 ± 0.02 0.21 ± 0.40a 0.29 ± 0.05b
Glutathione peroxidase 0.53 ± 0.09 0.51 ± 0.04 0.37 ± 0.04a 0.55 ± 0.03b
(U mg−1 protein)
SP, Spirulina; CsA, cyclosporine. Values are expressed as mean ± SD (n = 7). a P < 0.05 vs control; b P < 0.05 vs CsA.
448 M. KHAN ET AL.
Copyright © 2006 John Wiley & Sons, Ltd. J. Appl. Toxicol. 2006; 26: 444–451
DOI: 10.1002/jat
Figure 3. Histopathological examination of rat kidney (H&E 60×). (a) Control rat shows normal morphology; (b)CsA-treated rat shows severe isometric vacuolization (indicated by arrow) and widening of interstititum (c) CsA +Spirulina treated rat shows normal tubulointerstitial pattern. The results show that Spirulina protected the CsA-induced renal morphological alterations. This figure is available in colour online at www.interscience.wiley.com/journal/jat
Histomorphological Studies
Histological studies of rat kidney in the controls showed
a normal morphological appearance (Fig. 3a), whereas
the kidney of CsA-treated animals showed severe isomet-
ric vacuolization and widening of the interstitium
(Fig. 3b). Rats in the CsA + Spirulina group showed a
normal tubulointerstitial pattern with fewer isometric
vacuolizations (Fig. 3c).
Discussion
Cyclosporine therapy can lead to functional and structural
changes in the kidney of transplant patients and experi-
mental animals leading to renal dysfunction (Jackson
et al., 1987; Andoh et al., 1996). CsA treatment causes
a dose-related decrease in renal function in experimental
animals. The results of the present study showed that in
CsA-treated animals, there was a significant increase in
the plasma urea and creatinine, and a decrease in the
creatinine and lithium clearance. The increase in urinary
β-NAG activity in CsA treated animals indicates renal
tubular damage. These results supports our earlier obser-
vation (Kumar et al., 1999; Shifow et al., 2000) and
those of others (Whiting and Simpson, 1988) showing an
increase in urinary β-NAG activity in CsA treated ani-
mals. Pretreatment with Spirulina significantly attenuated
the CsA-induced nephrotoxicity. In Spirulina treated
animals, there was no impairment in renal function or
morphological changes induced by CsA. The mean
trough whole blood CsA levels in rats treated with CsA
alone was 2869 ± 334 ng ml−1, and in CsA + Spirulina
treated rats was 2468 ± 247 ng ml−1. These results suggest
that Spirulina offer protection against CsA-induced
nephrotoxicity without interfering in CsA metabolism.
The pathogenesis of the CsA-induced nephrotoxicity is
not fully understood, but it is thought to result from the
low-grade hypoxic injury to renal tubular cells (Andoh
et al., 1997), renal impairment of fibrogenic growth
factors, vasoconstrictor agents and reactive oxygen
species (Parra et al., 1998; Zhong et al., 1998). Recent
SPIRULINA ON CSA-INDUCED NEPHROTOXICITY 449
Copyright © 2006 John Wiley & Sons, Ltd. J. Appl. Toxicol. 2006; 26: 444–451
DOI: 10.1002/jat
studies have also demonstrated that CsA induces iNOS
and apoptosis in the renal tubular cells (Amore et al.,
1995).
A relationship between oxidative stress and
nephrotoxicity has been confirmed in many experimental
animals. Treatment with SOD and α-tocopherol signifi-
cantly reduced the nephrotoxic symptoms (Washio et al.,
1994). A role of oxygen free radical formation in
CsA-mediated impairment of renal function has been
suggested (Wolf et al., 1994). In the present study, CsA-
treated animals showed an increase in the MDA levels in
kidney tissue, and a decrease in antioxidant status.
Pretreatment with Spirulina attenuated the CsA-induced
increase in MDA levels and restored the antioxidant
enzymes to normal values. Previous studies have demon-
strated that treatment with a Spirulina enriched diet
increases the cerebellar glutathione levels, reduces MDA
levels and decreases the proinflammatory cytokines
(Bickford et al., 2000; Gemma et al., 2002). Several anti-
oxidants have been used to attenuate CsA-induced
nephrotoxicity (Tariq et al., 1999; Kumar et al., 1999;
Reiter et al., 2000; Padi and Chopra, 2002). Melatonin,
a potent hydroxyl radical scavenger, protected the CsA-
induced renal tubular damage (Kumar et al., 1999; Reiter
et al., 2000). Carvedilol, a beta-blocker with potent free
radical scavenger activity, reduced the MDA levels
and improved the renal dysfunction and morphological
changes induced by CsA (Padi and Chopra, 2002).
Co-administration of catechin with CsA significantly
reduced the lipid peroxidation and restored the decreased
glutathione levels induced by CsA (Anjaneyulu et al.,
2003).
In the present study, pretreatment with Spirulina sig-
nificantly attenuated the CsA-induced nephrotoxicity and
this effect is attributed to its antioxidant property. Re-
cently, it was demonstrated that Spirulina attenuated the
cisplatin-induced nephrotoxicity in rats and doxorubicin-
induced cardiotoxicity in mice (Mohan et al., 2006; Khan
et al., 2005). Several studies have shown that Spirulina
has potent antioxidant activity (Premkumar et al., 2001;
Upasani and Balaraman, 2003; Khan et al., 2005), and
is reported to possess hydroxyl and peroxyl radical
scavenging activity both in vitro and in vivo by
phycocyanin (Bhat and Madyastha, 2000). It has been
established that phycocyanin not only scavenges peroxyl,
hydroxyl (Vadiraja et al., 1998), peroxynitrite (Bhat
and Madhyastha, 2001) and superoxide radicals (Romay
et al., 1998), but also acts as a potent antioxidant and
inhibits lipid peroxidation (Romay et al., 1998; Gonzalez
et al., 1999; Romay and Gonzalez, 2000; Remirez et al.,
2002). Mice pretreated with Spirulina, significantly at-
tenuated the cyclophosphamide and mitomycin-C-induced
decrease in SOD, catalase, glutathione and glutathione-
dependent enzymes in the liver (Premkumar et al., 2001).
Recent studies have suggested that Spirulina protected
lead-induced toxicity in rats by inhibiting lipid
peroxidation and also by restoring endogenous antioxi-
dants (Upasani et al., 2001; Upasani and Balaraman,
2003). Spirulina was also shown to inhibit the zymosan-
induced arthritis in mice (Remirez et al., 2002). C-
phycocyanin selectively inhibits the pro-inflammatory
enzyme, cyclooxygenase-2 (COX-2) (Reddy et al., 2003).
Although the results obtained from the present study
suggest that Spirulina attenuated CsA-induced
nephrotoxicity through the inhibition of oxidative stress,
other possible mechanism(s) cannot be ruled out. Chronic
CsA administration may lead to induction of iNOS in rat
kidneys resulting in an increased production of nitric
oxide, leading to the formation of toxic peroxynitrite
(Amore et al., 1995; Amore and Coppo, 2000). Many
studies have suggested that nitric oxide and peroxynitrite
mediates renal cellular apoptosis leading to cellular dam-
age (Amore and Coppo, 2000). It is evident from the
many studies that apoptosis is involved in CsA-induced
renal injury (Sandau et al., 1997; Yang et al., 2002). A
recent study indicated that treatment with a Spirulina
enriched diet, reduced the ischemia-reperfusion induced
apoptosis and cerebral infarction by inhibiting caspase-3
activity (Wang et al., 2005). In our recent study, it was
demonstrated that Spirulina and C-phycocyanin signifi-
cantly inhibited the doxorubicin-induced free radical gen-
eration and apoptosis by attenuating caspase-3 activity in
isolated rat cardiomyocytes (Khan et al., 2006). It is
likely that Spirulina might be modulating the oxidative
stress-mediated apoptotic pathway, thereby inhibiting the
CsA-induced apoptosis. However this requires further
investigation.
In summary, this study demonstrated that Spirulina
significantly attenuated the CsA-induced nephrotoxicity
by reducing oxidative stress. Furthermore, Spirulina does
not interfere in the CsA metabolism. Therefore under-
standing the exact mechanism(s) of action of Spirulina
in reducing CsA-induced nephrotoxicity will be critical
in minimizing or preventing nephrotoxicity of CsA in
patients with CsA therapy. Thus, the nephroprotective
role of Spirulina against CsA-induced nephrotoxicity
needs further clinical evaluation.
Acknowledgement—We thank M/s Parry Neutraceuticals, Chennai forproviding pure powder of Spirulina for our study. Khan is presentlyworking as Post-doctoral fellow and Kutala is a Visiting Scientist at theOhio State University, Columbus, OH, USA
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