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


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.


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.



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.


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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



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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Spirulina attenuates cyclosporine-induced nephrotoxicity in rats

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