e-SPEN, the European e-Journal of Clinical Nutrition and Metabolism (2008) 3, e62ee71

e-SPEN,
the European e-Journal
of Clinical Nutrition
and Metabolism
http://intl.elsevierhealth.com/journals/espen

ORIGINAL ARTICLE

Garlic (Allium sativum) and ginger (Zingiber

officinale) attenuate structural nephropathy
progression in streptozotocin-induced
diabetic rats*
Khaled Al-Qattan*, Martha Thomson a, Muslim Ali b

Department of Biological Sciences, Faculty of Science, Kuwait University, P.O. Box 5969, Safat 13060, Kuwait

Received 7 April 2007; accepted 21 December 2007

KEYWORDS Summary
Hyperglycaemia; Background & aims: Diabetic nephropathy is a consequence of metabolic abnormalities spear-
Diabetes; headed by hyperglycemia. This study investigated the hypoglycaemic effects of garlic and gin-
Kidney; ger on the kidney structure of streptozotocin-induced diabetic rats.
Structural nephropathy; Method: Diabetic Sprague Dawley rats received a single daily intraperitoneal dose (500 mg/kg) of
Garlic; extracts of raw garlic (n Z 10) or ginger (n Z 10) for seven weeks. Normal rats (n Z 10) and non-
Ginger treated diabetic rats (n Z 10) acted as reference. At the last week, serum glucose and 24-h pro-
tein clearance were determined, and kidneys were processed for light microscopy examination.
Results: Compared to normal levels, non-treated diabetic serum glucose and protein clearance
were 330% and 185%, respectively (P < 0.05). Compared to non-treated diabetic rats, garlic or gin-
ger-treated diabetic rats serum glucose and protein clearance levels decreased by 45% and 50%,
respectively (P < 0.05). Images of non-treated diabetic kidneys showed characteristic histopath-
ological changes (e.g. capsular space shrinkage, glomerular hypertrophy and diffusion, glomerular
and microvascular eosinophilic precipitation, and cytoplasm fragmentation and retraction). In
garlic or ginger-treated rats, these renal structural changes although evident were less prominent.
Conclusions: The hypoglycaemic effects of garlic and ginger concurred with attenuation in the
progression of diabetic structural nephropathy.
ª 2008 European Society for Clinical Nutrition and Metabolism. Published by Elsevier Ltd.
All rights reserved.

*
Oral and poster presentation at the ‘International Diabetes Conference e 2006’, October 30eNovember 2, 2006 e Kuwait City, Kuwait.
* Corresponding author. Tel.: þ965 4987442, þ965 9611693; fax: þ965 4847054.
E-mail addresses: qattan@kuc01.kuniv.edu.kw (K. Al-Qattan), mmtkuwait2003@yahoo.com (M. Thomson), alimuslim@hotmail.com (M. Ali).
a
Tel.: þ965 4985710.
b
Tel.: þ965 4985706.

1751-4991/$ - see front matter ª 2008 European Society for Clinical Nutrition and Metabolism. Published by Elsevier Ltd. All rights reserved.
doi:10.1016/j.eclnm.2007.12.001
Effects of garlic and ginger on diabetic nephropathy
Introduction
It is reported that the incidence of diabetes is escalating at
an alarming rate and becoming a serious health and cost
issue to the point of being labeled an epidemic.1 It is also
predicted that most diabetics will experience some degree
of diminution in renal clearance at some stage following
onset of the condition.2 A deficiency in renal clearance
function is now confirmed as being the primary and/or
secondary initiator of a multitude of overt morbidities as
well as being a momentous contributor to high rates of
mortality.3
Type 1 and 2 diabetes mellitus represent typical forms of
endocrinal metabolic disorders. Because of insulin insuffi-
ciency and/or dysfunction, a diabetic suffers from impaired
glucose tolerance.3 The consequential hyperglycaemia was
previously viewed as the classical, and to a certain extent
the sole, biochemical indicator of diabetes. However,
recently additional biochemical and physiological abnor-
malities are taken as further hallmarks of the condition.
The foremost of these additional diagnostic symptoms
include hyperlipidaemia, atherosclerosis and hypertension.
Prolonged exposure to pathologically elevated, endoge-
nously produced biomolecules including oxidative/
nitrosative free radicals is a pivotal cause of tissue stress
and injury.4 The free radical permanent damage to tissue
structures results from an irreversible alteration in the mo-
lecular configuration of carbohydrates, lipids, proteins and
even nucleic acid bases.5 Change in the configuration of
principle structural biomolecules is regarded as a mediator
of the histoephysio-pathology of several medical disor-
ders.6 In diabetes, the level of free radicals was reported
to increase7 and in alloxan8 and STZ9 treated rats an
elevated level of free radicals was detected in several
tissues including the kidneys. The increased radical level
in diabetes could be attributed to a compromise in the
natural antioxidant regulatory mechanism and an increase
in free radical generation and flooding.10 Hyperglycemia
and a rise in glucose autooxidation are considered as major
inducers of oxidative stress and reported to be responsible
for micro- and macro-angiopathy that has been implicated
in the induction of an array of diseases including nephropa-
thy.11e16 The diabetic kidney demonstrates further
progressive structural alterations. Some of these signature
changes include tissue hyalinization, membrane thickening,
structure condensation and/or diffusion, and cytoplasmic
fragmentation in different segments of the nephron and
associated micro- and macro-vessels.17
Endogenously produced antioxidants are the body’s
chief mechanism for combating the destructive nature
of free radicals; nonetheless, the level of these free
radical scavengers could be supplemented by the intake
of foods rich is these agents. There is conclusive evidence
that both garlic and ginger possess numerous ameliorating
actions including induction of hypoglycemia.18e20 The
remedying potentials of these two Allium species are
ensued in their ability to enhance and/or repress certain
biochemical steps or pathways of correspondingly defec-
tive homeostatic mechanisms. It is highly probable that
the reported corrective potentials of garlic and ginger
could also be due to their high contents of antioxidants
e63
as well as increasing antioxidants concentrations upon
administration in vivo.21e24 This enhanced bioavailability
of antioxidants brought about by garlic and ginger could
be crucial in alleviating oxidative/nitrosative injury of
tissues.24 Since garlic and ginger can amend pathological
metabolic changes, it is highly probable that they can pro-
tect against structural changes that develop in diseased
conditions.8,25 More specifically, garlic and ginger might
reduce the occurrence and/or progression of structural
nephropathies and, therefore, delay the onset of renal
failure and ultimately the end-stage of the disease in
diabetes.25
There is support to the premise that active ingredients in
herbs express their therapeutical actions most effectively
when they work synergistically. Therefore, the general
trend now when using natural products for medicinal
application is to use them in their natural available form.
This study investigated the hypoglycemic effects of whole
extracts of garlic and ginger on the development of typical
renal structural changes in diabetic rats. There is
compelling support to the probability that maintaining
serum glucose comparable to euglycaemic levels is vital
to protecting kidney structure.26,27 Streptozotocin (STZ)-
induced hyperglycaemic rats were produced and used as
an experimental model of type I diabetes.9
Materials and methods
Animals
The animals used in this study were male Sprague Dawley
rats (body weight: 180e200 g e parents: B&K Universal
Company, Hull e England). The rats were housed in the
animal care facility (Department of Biological Sciences e
Kuwait University) under mild ambient climatic conditions
and provided with standard rodent diet plus tap water ad
libitum. The rats were maintained, then, euthanized
according to the guidelines of the Laboratory Animals
Care Protocols.
Induction of hyperglycaemia
Over 40 rats were injected intraperitoneally with a single
dose of 60 mg STZ/kg/0.5 ml normal saline following an
overnight fast. Simultaneously, 10 additional rats received
a comparable dose of normal saline and were considered
as normal rats (N).
Collection of blood samples and determination
of serum glucose levels
Blood samples (0.5 ml) were drawn from all rats via cardiac
puncture under diethyl ether mild sedation one-week post-
treatment with STZ or normal saline. Isolated serum was
assayed for glucose concentration using the glucose oxidase
method (CARO Co. e Germany). STZ-injected rats with
serum glucose concentrations higher than 350 mg/dl were
designated diabetics (D) and used in the study (n Z 30).
Further blood samples were collected after the seven
weeks and assayed for glucose concentration.
e64
Treatments of hyperglycaemic rats
with normal saline, garlic or ginger
One-week post-STZ injection the D rats were randomly
divided into three groups (3  10) and immediately each
group started receiving a different treatment as a single
daily intraperitoneal injection of 0.5 ml for a period of
seven weeks: 1 e normal saline (D); 2 e garlic extract (500
mg/kg) (D þ GC); 3 e ginger extract (500 mg/kg) (D þ GR).
The N rats received a comparable dose of normal saline
for the same course of treatment (N þ NS).
Determination of urine protein clearance
All rats were placed in metabolic cages for urine sample
collection during the seventh week of treatment. Daily
urine samples were centrifuged to remove any food
particles. The urine protein levels were then determined
using the Coomassie Brilliant Blue method. This method was
used to estimate the total daily renal clearance of protein,
which is taken as a marker of severe damage of glomerular
filtration barrier.
Preparation of kidney tissue for light
microscopy examination and imaging
After completion of treatments and following euthanasia
with sodium pentobarbital (Sagatal, May & Baker e
England), 2-mm-thick cross sections of the left kidneys
of three arbitrarily selected rats from each group were
prefixed by immersion into 4% formalsaline solution for
2 h, then carried through standard Bowan fixation and
Eosinehematoxylin staining. About 20 (5 mm thick) renal
sections from each rat in each group (60 sections/group)
were prepared and examined under light microscopy
(Olympus Vanox AHBT3, Tokyo e Japan). Digital images
of specific renal regions and structures were acquired in
a blind fashion from the 60 specimens for each group us-
ing a fitted digital camera (Olympus DP70, Tokyo e Japan)
connected to a computer loaded with digital camera
software interface (OLYSIA BioAutoCell, Tokyo e Japan).
The width of the renal corpuscles capsular space (RC-CS)
was estimated using a microscope eyepiece graticule.
For each experimental group 100 readings were
estimated. Each reading of the 100 readings is an average
of four measurements taken as 2  2 readings along two
perpendicular axes running through the glomerulus. Each
one of the four readings represents the distance between
the external surface of the glomerulus and the opposite
inner surface of the capsular membrane. The two
Table 1 Serum glucose concentration and urine protein clearance at the seventh week
N D
Serum glucose (mg/dl) 129  9 (n Z 10)
Urine protein (mg/24 h) 7  1 (n Z 10)
Normal rats (N), diabetic rats (D), garlic-treated diabetic rats (D þ GC) and ginger-treated diabetic rats (D þ GR).
a
Significantly different compared to normal.
b
Significantly different compared to control diabetic.
K. Al-Qattan et al.
perpendicular axes were parallel to the length axis and
width axis of the whole kidney.
Statistical analysis D data and images
presentation
Seventh week serum glucose and urine protein clearance
are presented in Table 1 as means  SEM, while the average
width of the renal corpuscle capsular space is shown in
a bar graph representing means  SEM (Fig. 6). Readings
of N, D and D þ GC or N, D and D þ GR were compared using
one-way ANOVA (SPSS, v. 10). Differences were considered
significant when P < 0.05. Five digital images combinations
(DIC) showing renal cortices (DIC-1 e Fig. 1), renal
corpuscles (DIC-2 e Fig. 2), renal proximal tubules (DIC-3
e Fig. 3), renal medullae (DIC-4 e Fig. 4), and renal arcuate
arteries (DIC-5 e Fig. 5) are presented as black & white
prints. Each combination displays four images of the same
renal region or structure and each image corresponds to
one of the four groups of rats (N, D, D þ GC, and D þ GR).
Results
Biochemical data
The serum glucose reading of the N rats was normal at
129  9 mg/dl. In contrast, the serum glucose concentra-
tion of the D rats was pathologically elevated at
431  13 mg/dl and more than threefold higher (P < 0.5)
than that of the N-NS. The D þ GC and D þ GR glucose
values at 237  8 and 267  23, respectively, were less by
45% (P < 0.5) than that of the D. Although the D þ GC and
D þ GR serum glucose readings were almost twice those of
the N rats, they were not significantly different. The urine
protein indicates that the daily protein clearance of the D
rats at 13  2 was twofold higher (P < 0.5) than that of
the N rats. The D þ GC and D þ GR protein clearances at
6  1 and 5  1 were lower by 50% (P < 0.5) than that of D
rats and almost equal to the protein clearance of the N rats.
Fig. 6 indicates that the average width of the RC-CS in
the D rats was less by 45% (P < 0.5) than that in the N
rats. The D þ GC had a wider (P < 0.5) RC-CS compared to
that of the D rats; however, it was still narrower (P < 0.5)
than that in the N rats. As for the D þ GR, the width of
the RC-CS was less than that in the N rats (P < 0.5) and
not different from the RC-CS in the D rats.
Histological data
The images of each experimental group were acquired
from different sections of renal tissue representing the
D þ GC D þ GR
a b
431  1 (n Z 10) 237  (n Z 10) 267  2b (n Z 10)
13  2a (n Z 10) 6  1b (n Z 10) 5  2b (n Z 10)
Effects of garlic and ginger on diabetic nephropathy
Figure 1 DIC-1: cortices (CX e 10) of normal kidney (N-CX), diabetic kidney (D-CX), garlic-treated diabetic kidney (D-CX þ GC),
and ginger-treated diabetic kidney (D-CX þ GR). RC Z renal corpuscle, PT Z proximal tubule, DT Z distal tubule, CT Z collecting
tubule, TL Z tubular lumen, NS Z nucleus, PS Z protein shedding, TNS Z thinning in nucleus, CNS Z condensation in nucleus.
arbitrarily selected rats. These images, when considered
collectively, provided an overall picture of the type of
structural change and status of the renal tissue in each
experimental group.
Examination of the digital images combination (DIC-1 e
Fig. 1) of the renal cortices (CX) at low magnification
(10) revealed the following: Compared to the CX of the
N rats the CX of D rats had a similar overall architecture.
However, the majority of the renal corpuscles (RC) in the
D rats’ kidneys appeared to have distorted capsular shape
that reflected on the form of the glomeruli. Furthermore,
most of the RC-CS in the D rats’ kidney showed the
presence of protein shedding (capsular drops) that may
represent insudation of the plasma proteins. Although
these protein markers are evident in the N rats’ capsular
space (CS), they were more prevalent in the RC-CS of D
rats. The general form of the proximal tubule (PT), distal
tubule (DT) and collecting duct (CD) appear normal at this
magnification.
The CX of the D þ GC and D þ GR showed RC with more
rounded shape almost similar to that seen in the CX of
the N rats. Nonetheless, some structural irregularities are
still evident in the RC of the D þ GC and D þ GR. The PT
and DT general shapes are consistent with those seen in
the kidney of N rats. The nuclear appearance is different
between the D þ GC and D þ GR, were the nuclei in the
e65
D þ GC still looked larger with some chromatin thinning,
while those of the D þ GR appeared denser.
Meticulous examination of the RC at the magnification of
40 (DIC-2 e Fig. 2) revealed that the RC of the D rats
showed early signs of irregular form, considerable shrink-
age in the CS to the point where in some places and along
a good length the capsular wall touched the glomerulus.
The capsular wall at certain segments lost definition to
the point of distortion, whereas at other places some thick-
ening of the basement membrane was obvious. The glomer-
ulus of the D rats’ kidney showed signs of hypertrophy and
uneven distribution with more condensation of mesangial
cells and capillary epithelial cells and in some places fusion
of mesangial cells. An interesting glomerular structural
change present in the D rats’ kidney was streaks of conden-
sations that radiated through the glomerular bed originat-
ing and extending from the vascular pole. In the kidneys
of the D þ GC or D þ GR, and compared to the kidney of
the D rats, the RC showed better corpuscular form (oval
to round), capsular wall integrity, capsular space dimen-
sions, glomerular layout and nuclear shape. Furthermore,
after garlic or ginger treatments there were less condensa-
tion deposits, mesangial fusion, and protein shedding.
The PT in kidneys of the D rats showed several alterations
indicating distinct changes compared to the structure of the
PT in the kidney of N rats. In the D rats’ kidney, the PT
e66
Figure 2 DIC-2: renal corpuscles (RC e 40) of normal kidney (N-RC), diabetic kidney (D-RC), garlic-treated diabetic kidney
(D-RC þ GC), and ginger-treated diabetic kidney (D-RC þ GR). CWD Z capsular wall distortion, MF Z mesangial fusion, CS Z capsular
space, CW Z capsular wall, GC Z glomerular condensation, MVC Z micro-vascular condensation.
basement membranes were distorted in certain places and
thick in others. The cytoplasm was fragmented and basely
retracted. Most of the nuclei appeared enlarged and demon-
strated chromatin thinning. The tubular lumens showed
sizable light globular formations surrounded by dark borders
in shades of gray. The peritubular capillary vessels around
the PT appeared enlarged and dilated.
In comparison to the PT of the D rats, the PT of the
D þ GC generally had more normal cell distribution and
architecture. In addition, the PT of the D-GC had thinner
basement membranes that nicely supported and made
good contact with the cytoplasm of good sized and intact
cuboidal epithelial cells that had well-rounded nuclei.
The cytoplasm of these cells showed lesser fragmentation,
but exhibited higher intensity of staining. The tubular
lumens were smaller and in some cases, the lumen was
no more than a radiating center lined by the brush border.
As is the case with the PT of the three other groups of rats,
the PT of the D-GR showed a normal gross architecture. The
PT of the D þ GR are well defined by a slightly thicker
basement membrane compared to the PT of the N þ NS
and D þ GC, but are slightly thinner than those seen in
the PT of the D þ NS. The PT of the D þ GR are internally
lined with normal size cuboidal epithelial cells that made
good contact with the basement membrane and surround
mostly centrally positioned normal and darkly stained
K. Al-Qattan et al.
nuclei. The cytoplasm of the PT in the D þ GR seemed
intact compared to that seen in the PT of D rats; however
it showed signs of granulation. The lumens in the D þ GR
were larger than those seen in the PT of the D þ GC, but
were slightly smaller than those observed in the PT of the
N and D rats.
The digital image of the upper medullary region (DIC-4
eFig. 4) suggested that the D þ NS had normal distribution
of Henle’s tubules. However, detailed examination of these
tubules reveal that they have slightly thicker basement
membranes and loss of tubular definition of the cellular
and luminal form, in addition to cytoplasm fragmentation
in both the ascending and descending limbs. The nuclei of
the epithelial cells in the ascending tubules appear as
darkly stained dots that concentrate closer to the center.
The vasa recta vessels are more pronounced with thicker
basement membranes and larger nuclei compared to those
in the normal vasa recta. Furthermore, the D rats’ vasa
recta borders are rougher in appearance than those noticed
in the N rats.
The upper medulla of the D þ GC kidney showed a typical
layout of descending and ascending limbs of the loop and
other structures. Compared to the D rats’ kidney, the limbs
in the kidney of the D þ GC had better definition of cuboidal
cells and evenly distributed, slightly denser cytoplasm. The
nuclei were well defined and centrally located, but showed
Effects of garlic and ginger on diabetic nephropathy
Figure 3 DIC-3: proximal tubules (PY e 40) of normal kidney (N-PT), diabetic kidney (D-PT), garlic-treated diabetic kidney
(D-PT þ GC), and ginger-treated diabetic kidney (D-PT þ GR). BB Z brush boarder, BM Z basement membrane, RBM Z retracted
basement membrane, TBM Z thick basement membrane, DBM Z distorted basement membrane, GF Z globular formation,
RBM Z retracted basement membrane, CP Z cytoplasm.
signs of hypertrophy and thinning. The vasa recta in the
D þ GC kidney still showed signs of distorted apical cellular
lining. The tubules had better-defined lumens; however,
less defined basement basal membranes. The upper
medulla in the kidney of the D þ GR showed similar struc-
tural features compared to those seen in the D þ GC upper
medulla. However, in the tubules of D þ GR there are signs
of better definition of basal membranes and fewer enlarged
nuclei. The cuboidal cells in the limbs of the kidney in the
D þ GR were less defined compared to those observed in the
D þ GC tubules.
The digital images of the renal arcuate arteries (RAA)
(DIC-5 e Fig. 5) showed that the artery of the D rats had the
same form as that present in the N rats. However, the RAA
of the D rats showed two distinct structural differences
exemplified in the presence of a thicker internal elastic
lamina and the presence of more batches of intense
coloration in the tunica media. The thickness of the inter-
nal elastic lamina in the RAA of the D þ GC appears to be
between those of the N rats and the D rats, whereas, the
RAA of the D þ GR seems to be as thin as that found in
the N rats. The tunica media of the RAA in the D þ GC
and D þ GR still showed some intense coloration; nonethe-
less, the intensity of this coloration was slightly less. The
condition of the endothelium in the RAA of all groups could
not be ascertained at a magnification of 40.
e67
Discussion
The etiologies of serious and ultimately fatal diseases
inherent to diabetes are distinct biochemical and
physiological abnormalities. It is almost certain that
chronic increases in the concentration of glucose creates
an internal milieu of elevated free radicals that imposes
deleterious stress leading to permanent histological
damage. This study focused on two of the most established
defects that afflict diabetics, namely hyperglycaemia and
structural nephropathy, and investigated the possibility
that the hypoglycaemic actions of garlic and ginger might
attenuate the intensity of structural nephropathy in STZ-
induced diabetic rats.
In a review, Ceriello16 stated that hyperglycaemia-
induced overproduction of superoxide seems to be the first
and main event in the activation of all pathways involved in
the pathogenesis of diabetes complications. Furthermore,
increased generation of nitric oxide and, consequently
the formation of the strong oxidant peroxynitrite accompa-
nied superoxide overproduction. In another study,24 it was
reported that STZ-injected rats displayed variable changes
in homeostatic levels of glucose, total lipids, triglyceride,
cholesterol lipid peroxides, nitric oxide and uric acid.
Concomitantly, significant decreases in the levels of the
antioxidants ceruloplasmin, albumin and total thiols were
e68
Figure 4 DIC-4: renal medullae (RM e 40) of normal kidney (N-RM), diabetic kidney (D-RM), garlic-treated diabetic kidney
(D-RM þ GC), and ginger-treated diabetic kidney (D-RM þ GR). VR Z vasa recta.
detected in the plasma of these diabetic animals. It is clear
from the previous reports that the concentrations of power-
ful oxidative/nitrosative free radicals increase while those
of antioxidants decrease in the circulation of diabetics. As
a result, the amounts of radical agents transported in the
blood to different tissues and organs of the body become
pathologically elevated.
Both kidneys receive an estimated 20% of the cardiac
output, which is high considering that their combined
weight does not exceed 0.5% of the total weight of the
body. In diabetes, as large volumes of blood are delivered
with each heartbeat, the kidneys become perfused with
large amounts of radical toxins.11 The renal exposure to
toxins is augmented further by the kidneys’ own production
of free radicals which becomes elevated in condition of
diabetes. As such, because of the exposure to free radicals
from both extrinsic and intrinsic sources, the kidneys are
regarded as a primary target to the deleterious effects of
these metabolites.28,29 The histological observations of
the present study provide support to the destructive nature
of these radicals on the diabetic kidneys. The histological
images of the diabetic normal saline-treated kidneys show
clear signs of structural distortions that developed after
a short period of hyperglycaemia and were obvious even
under the low magnification of light microscopy. Early
changes in tissues structure have also been reported in
the diabetic cardiovascular system materializing in the
K. Al-Qattan et al.
form of coronary calcifications.11 It is suggested that most
of these changes result from membranous distortions that
could be due to peroxidations of membrane lipids and
damage to membrane protein structure extending from
the primary to quaternary level.
In this study, the terminal values of serum glucose and
urine protein of the four groups of rats shown in the table
(and when compared to each other) present a comprehen-
sive conclusion regarding the type of changes in the levels
of these two parameters over the course of the seven
weeks of the study. This is supported by detailed periodic
measurements and analysis of serum glucose and urine
protein, in addition to the body weight, water intake and
urine output (þ serum levels of triglyceride and
cholesterol) reported in two recent and related studies in
the same groups of rats.20,30 In addition to improvements
in serum glucose level and urine protein clearance in these
two studies, the data show that treating diabetic rats with
either garlic or ginger led to: (a) stabilization in the body
weight of the treated animals close to the initial weight
and (b) reduction in water consumption and urine output
toward normal levels. When the body weight, water
consumption and urine output data of the three diabetic
groups are collectively considered, they suggest that the
reported beneficial effects of ginger and garlic are not
due to a change in diet. This conclusion is supported by
the finding that the significantly higher body weight of
Effects of garlic and ginger on diabetic nephropathy e69

Figure 5 DIC-5: renal arcuate arteries (RAA e 40) of normal kidney (N-RAA), diabetic kidney (D-RAA), garlic-treated diabetic
kidney (D-RAA þ GC), and ginger-treated diabetic kidney (D-RAA þ GR). IEL Z Internal elastic lamina, TM Z Tunica media,
RBC Z red blood cells.

the treated diabetic rats (compared to the non-treated
diabetics) was associated with reduction in both serum
glucose and urine protein clearance.
Most of the structural changes observed in the diabetic
kidneys treated with normal saline were associated with
the different segments of the nephron and in particular the
components of the RC and associated arterioles. It appears
that the glomerulus is a principal site for the action of
as proteases, which particularly damage and digest the glo-
merular filtration barrier.32 The deteriorating effects that
the glomerulus experienced is further indicated by these
observations: (1) the average width of the CS in the dia-
betic RC was significantly reduced in comparison to that
measured in the normal RC, which indicates a reduction
renal corpuscle space average

reactive oxygen species leading to glomerulonephritis.31 1

The structural changes included distortion and thickening 0.9
in the capsular membranes, shrinkage in the capsular 0.8
space, uneven condensation, diffusion and hypertrophy of 0.7
width (Um)

0.73
0.6 a
the glomerular body. The most striking feature noticed b
0.5 a
was streaks of condensation originating from the vascular a
0.4 0.46 0.43
poles and radiating throughout the glomerulus in the kidney 0.3
0.39
of normal saline-treated hyperglycaemic rats, which could 0.2
have resulted from the deposition of eosinophilic mate- 0.1
rial.17 The formation of this material was considered as 0
a prelude to the hyalinization of the glomerulus that is N D D+GC D+GR
characteristic of diabetic glomeruli. The structural changes a
: significantly different compared to normal rats.
in the glomerulus are clear signs of glomerulonephritis that b
: significantly different compared to control diabetic rats.
substantially compromises the filtration integrity of the
glomerulus due to damage and dysfunction of endothelial Figure 6 Renal corpuscle capsular space average width (mm)
cells11 and podocytes.12 These structural and functional de- in normal rats (N), saline-treated diabetic rats (D), garlic-
fects of the glomerulus may be aggravated further by the treated diabetic rats (D þ GC) and ginger-treated diabetic rats
glomerular cells’ own production of agents such as eicosa- (D þ GR). a: Significantly different compared to normal rats. b:
noids, chemokines, cytokines, and growth factors, as well Significantly different compared to control diabetic rats.
e70
in filtration rate and (2) the daily clearance of protein was
significantly elevated indicating proteinuria, a parameter
established as a marker of diabetic nephropathy.33
It is well known that free radicals by virtue of their
ability to extract or donate electrons change the valence
characteristics of affected atoms. This may lead to altering
atomic reactivity, modulating molecular structure, hence
causing abnormalities in the configuration of major classes
of structural biomolecules. It is expected that controlling
the endogenous generation and bioavailability of free
radicals will preserve normal atomic physical and chemical
properties and thus reactivity. The view that consumption
of antioxidant-rich foods is important in combating disease
is receiving acceptance and support.34 Garlic and ginger
have been shown to possess the potential to effectively
ameliorate a considerable number of diabetic biochemical
and physiological modulations targeted in investigative
studies. In addition, the amounts of garlic and ginger used
in this study were reported to have no damaging effects
on the structures of tissues such as lung or liver in normal
rats.35,36 These corrective actions have been suggested to
be partially due to their content, as other plants species,
of several biologically active compounds that possess
antioxidant properties.34 In the present study, the serum
glucose data showed clearly that garlic and ginger restrain
the level of hyperglycaemia resulting from the experimen-
tal destruction of beta pancreatic cells induced by STZ. The
hypoglycaemic actions of garlic and ginger appeared to
attenuate tissue injury that occurred in the kidneys of
treated diabetic rats which is suggested to be due to a re-
duction in the level of oxidants.24 The digital images of
the different regions and structures taken from the kidneys
of STZ-injected rats treated with garlic or ginger support
the previous claim. The nephrons of kidneys from treated
diabetic rats showed a healthier form in particular in the
glomerular region and the surrounding CS. These images
suggest lesser intensity of glomerulonephritis than that
observed in the kidneys of diabetic rats treated with normal
saline. The signs of lesser protein shedding observed in the
CS of the garlic or ginger-treated rats and the reduction in
the level of proteinuria confirm that the glomeruli in these
animals are performing much better than the glomeruli of
control diabetic rats. In support, it was previously reported
that garlic and ginger reduce albuminuria, hypertrophy and
collagen deposition in the glomerulus.8
The distortion in the PT of the hyperglycaemic rats
treated with normal saline suggest further that the
increased proteinuria observed in these animals could be
due to the impairment of tubular transport mechanisms
dedicated to processing and reclaiming filtered amino acids
and proteins. This suggestion is supported by the presence
of globular forms in the lumens of PT in non-treated STZ
rats. Because the PT are a bidirectional venue of reabsorp-
tion and secretion, its cells are susceptible to the injurious
effects of many oxidative/nitrosative ions and molecules
found in the glomerular filtrate passing through the PT and
the surrounding interstitial fluids.32 As a result, cellular
biomolecules, especially proteins, may become damaged
resulting in a distortion of cytoplasmic integrity and form
in that part of the nephron.5 Cytoplasmic fragmentation,
a main structural feature of diabetic PT pathology,32 has
also been observed in this study in the control STZ-kidney.
K. Al-Qattan et al.
This structural cytoplasmic change will diminish the PT
reabsorptive capacity allowing a considerable quantity of
filtered load of amino acids and proteins to pass into the
urine unclaimed by the diabetic kidneys. This reduction in
PT reabsorptive function may be aggravated by (1) damage
to the absorptive membrane due to loss of microvilli
observed in this study and reported elsewhere21 and (2)
destruction of the mitochondria of PT epithelia, which are
important for providing the energy necessary for active
transport mechanisms. In addition, it is reported that dam-
aged mitochondria release large amounts of reactive agents
that expose the diabetic kidneys to more of these deleteri-
ous agents.37 In this study, the injurious effects that the
diabetic kidneys are exposed were further highlighted in
the form of cytoplasmic distortion in the upper parts of
the renal loops located in the upper medullary region.
The argument to the cause of this damage is similar to
that presented for the PT. The garlic or ginger-treated
rats showed lower incidences of change in this segment of
the nephron.
The vascular endothelium has been reported to synthe-
size and release a wide range of vasoactive substances,
including the vasodilator NO, prostacyclin and endothe-
lium-derived hyperpolarizing factors, in addition to the
vasoconstrictors angiotensin II and endothelin-1.14 The or-
chestrated function of these vasoactive components under
normal physiological conditions is imperative to maintain-
ing normal blood perfusion and circulation. However, these
vasoactive agents can alter the circulation following endo-
thelial dysfunction that occurs under circumstances such
as trauma, hyperglycaemia or dyslipidaemia.14 Endothelial
dysfunction may result from membranous glycation.38 In
the present study, at the level of light microscopy,
evidence of glycation might be suggested by the distortion
of the endothelial cells of the glomerular vessels, the rough
appearance of the internal elastic lamina in the RAA, and
the rough appearance of the vasa recta in the non-treated
diabetic kidneys. Controlling the levels of serum glucose in
garlic and ginger STZ-injected rats ensured the preserva-
tion of the structural integrity of the micro- and macro-
vessels as suggested by the normal looking glomerular
capillary, elastic lamina of RAA and vasa recta of garlic
or ginger-treated kidneys. Thus, in response to these
protective effects of garlic and ginger, it is highly probable
that the intrinsic humoral regulatory functions of the
endothelium of the renal vasculature was maintained as
supported by the improved protein clearance functions.
It is concluded from the findings of this study that: (1)
the distortions in the structure of several nephronal and
microvascular components of the kidney materialize in
the early stages of diabetes; (2) the degree of renal
structural injury parallels the level of serum glucose; and
(3) garlic or ginger attenuates the progression of struc-
tural nephropathy.
Acknowledgments
Dr. Khaled Al-Qattan, Dr. Martha Thomson and Professor
Muslim Ali presented the idea for this investigation,
designed the investigation protocol, induced diabetes,
collected and analyzed samples, carried out histological
Effects of garlic and ginger on diabetic nephropathy
preparation, examination and capturing of digital images of
renal tissue sections, performed statistical analysis, carried
out literature search and cross-referencing, and wrote the
article.
We like to thank Mrs. Zainab Al-Amin for assisting in the
induction of diabetes and determination of serum glucose
and urine protein levels, Ms. Dalal Al-Hajeri for assisting in
the preparation of tissue sections and measurement of
renal corpuscle capsular space, and Kuwait University for
partially supporting this study.
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