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: [email protected] (K. Al-Qattan), [email protected] (M. Thomson), [email protected] (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 e63

Introduction as well as increasing antioxidants concentrations upon

administration in vivo.21e24 This enhanced bioavailability
It is reported that the incidence of diabetes is escalating at of antioxidants brought about by garlic and ginger could
an alarming rate and becoming a serious health and cost be crucial in alleviating oxidative/nitrosative injury of
issue to the point of being labeled an epidemic.1 It is also tissues.24 Since garlic and ginger can amend pathological
predicted that most diabetics will experience some degree metabolic changes, it is highly probable that they can pro-
of diminution in renal clearance at some stage following tect against structural changes that develop in diseased
onset of the condition.2 A deficiency in renal clearance conditions.8,25 More specifically, garlic and ginger might
function is now confirmed as being the primary and/or reduce the occurrence and/or progression of structural
secondary initiator of a multitude of overt morbidities as nephropathies and, therefore, delay the onset of renal
well as being a momentous contributor to high rates of failure and ultimately the end-stage of the disease in
mortality.3 diabetes.25
Type 1 and 2 diabetes mellitus represent typical forms of There is support to the premise that active ingredients in
endocrinal metabolic disorders. Because of insulin insuffi- herbs express their therapeutical actions most effectively
ciency and/or dysfunction, a diabetic suffers from impaired when they work synergistically. Therefore, the general
glucose tolerance.3 The consequential hyperglycaemia was trend now when using natural products for medicinal
previously viewed as the classical, and to a certain extent application is to use them in their natural available form.
the sole, biochemical indicator of diabetes. However, This study investigated the hypoglycemic effects of whole
recently additional biochemical and physiological abnor- extracts of garlic and ginger on the development of typical
malities are taken as further hallmarks of the condition. renal structural changes in diabetic rats. There is
The foremost of these additional diagnostic symptoms compelling support to the probability that maintaining
include hyperlipidaemia, atherosclerosis and hypertension. serum glucose comparable to euglycaemic levels is vital
Prolonged exposure to pathologically elevated, endoge- to protecting kidney structure.26,27 Streptozotocin (STZ)-
nously produced biomolecules including oxidative/ induced hyperglycaemic rats were produced and used as
nitrosative free radicals is a pivotal cause of tissue stress an experimental model of type I diabetes.9
and injury.4 The free radical permanent damage to tissue
structures results from an irreversible alteration in the mo- Materials and methods
lecular configuration of carbohydrates, lipids, proteins and
even nucleic acid bases.5 Change in the configuration of Animals
principle structural biomolecules is regarded as a mediator
of the histoephysio-pathology of several medical disor- The animals used in this study were male Sprague Dawley
ders.6 In diabetes, the level of free radicals was reported rats (body weight: 180e200 g e parents: B&K Universal
to increase7 and in alloxan8 and STZ9 treated rats an Company, Hull e England). The rats were housed in the
elevated level of free radicals was detected in several animal care facility (Department of Biological Sciences e
tissues including the kidneys. The increased radical level Kuwait University) under mild ambient climatic conditions
in diabetes could be attributed to a compromise in the and provided with standard rodent diet plus tap water ad
natural antioxidant regulatory mechanism and an increase libitum. The rats were maintained, then, euthanized
in free radical generation and flooding.10 Hyperglycemia according to the guidelines of the Laboratory Animals
and a rise in glucose autooxidation are considered as major Care Protocols.
inducers of oxidative stress and reported to be responsible
for micro- and macro-angiopathy that has been implicated Induction of hyperglycaemia
in the induction of an array of diseases including nephropa-
thy.11e16 The diabetic kidney demonstrates further
Over 40 rats were injected intraperitoneally with a single
progressive structural alterations. Some of these signature
dose of 60 mg STZ/kg/0.5 ml normal saline following an
changes include tissue hyalinization, membrane thickening,
overnight fast. Simultaneously, 10 additional rats received
structure condensation and/or diffusion, and cytoplasmic
a comparable dose of normal saline and were considered
fragmentation in different segments of the nephron and
as normal rats (N).
associated micro- and macro-vessels.17
Endogenously produced antioxidants are the body’s
chief mechanism for combating the destructive nature Collection of blood samples and determination
of free radicals; nonetheless, the level of these free of serum glucose levels
radical scavengers could be supplemented by the intake
of foods rich is these agents. There is conclusive evidence Blood samples (0.5 ml) were drawn from all rats via cardiac
that both garlic and ginger possess numerous ameliorating puncture under diethyl ether mild sedation one-week post-
actions including induction of hypoglycemia.18e20 The treatment with STZ or normal saline. Isolated serum was
remedying potentials of these two Allium species are assayed for glucose concentration using the glucose oxidase
ensued in their ability to enhance and/or repress certain method (CARO Co. e Germany). STZ-injected rats with
biochemical steps or pathways of correspondingly defec- serum glucose concentrations higher than 350 mg/dl were
tive homeostatic mechanisms. It is highly probable that designated diabetics (D) and used in the study (n Z 30).
the reported corrective potentials of garlic and ginger Further blood samples were collected after the seven
could also be due to their high contents of antioxidants weeks and assayed for glucose concentration.
e64 K. Al-Qattan et al.

Treatments of hyperglycaemic rats perpendicular axes were parallel to the length axis and
with normal saline, garlic or ginger width axis of the whole kidney.

One-week post-STZ injection the D rats were randomly Statistical analysis D data and images
divided into three groups (3  10) and immediately each presentation
group started receiving a different treatment as a single
daily intraperitoneal injection of 0.5 ml for a period of Seventh week serum glucose and urine protein clearance
seven weeks: 1 e normal saline (D); 2 e garlic extract (500 are presented in Table 1 as means  SEM, while the average
mg/kg) (D þ GC); 3 e ginger extract (500 mg/kg) (D þ GR). width of the renal corpuscle capsular space is shown in
The N rats received a comparable dose of normal saline a bar graph representing means  SEM (Fig. 6). Readings
for the same course of treatment (N þ NS). 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
Determination of urine protein clearance significant when P < 0.05. Five digital images combinations
(DIC) showing renal cortices (DIC-1 e Fig. 1), renal
All rats were placed in metabolic cages for urine sample corpuscles (DIC-2 e Fig. 2), renal proximal tubules (DIC-3
collection during the seventh week of treatment. Daily e Fig. 3), renal medullae (DIC-4 e Fig. 4), and renal arcuate
urine samples were centrifuged to remove any food arteries (DIC-5 e Fig. 5) are presented as black & white
particles. The urine protein levels were then determined prints. Each combination displays four images of the same
using the Coomassie Brilliant Blue method. This method was renal region or structure and each image corresponds to
used to estimate the total daily renal clearance of protein, one of the four groups of rats (N, D, D þ GC, and D þ GR).
which is taken as a marker of severe damage of glomerular
filtration barrier. Results

Biochemical data
Preparation of kidney tissue for light
microscopy examination and imaging The serum glucose reading of the N rats was normal at
129  9 mg/dl. In contrast, the serum glucose concentra-
After completion of treatments and following euthanasia tion of the D rats was pathologically elevated at
with sodium pentobarbital (Sagatal, May & Baker e 431  13 mg/dl and more than threefold higher (P < 0.5)
England), 2-mm-thick cross sections of the left kidneys than that of the N-NS. The D þ GC and D þ GR glucose
of three arbitrarily selected rats from each group were values at 237  8 and 267  23, respectively, were less by
prefixed by immersion into 4% formalsaline solution for 45% (P < 0.5) than that of the D. Although the D þ GC and
2 h, then carried through standard Bowan fixation and D þ GR serum glucose readings were almost twice those of
Eosinehematoxylin staining. About 20 (5 mm thick) renal the N rats, they were not significantly different. The urine
sections from each rat in each group (60 sections/group) protein indicates that the daily protein clearance of the D
were prepared and examined under light microscopy rats at 13  2 was twofold higher (P < 0.5) than that of
(Olympus Vanox AHBT3, Tokyo e Japan). Digital images the N rats. The D þ GC and D þ GR protein clearances at
of specific renal regions and structures were acquired in 6  1 and 5  1 were lower by 50% (P < 0.5) than that of D
a blind fashion from the 60 specimens for each group us- rats and almost equal to the protein clearance of the N rats.
ing a fitted digital camera (Olympus DP70, Tokyo e Japan) Fig. 6 indicates that the average width of the RC-CS in
connected to a computer loaded with digital camera the D rats was less by 45% (P < 0.5) than that in the N
software interface (OLYSIA BioAutoCell, Tokyo e Japan). rats. The D þ GC had a wider (P < 0.5) RC-CS compared to
The width of the renal corpuscles capsular space (RC-CS) that of the D rats; however, it was still narrower (P < 0.5)
was estimated using a microscope eyepiece graticule. than that in the N rats. As for the D þ GR, the width of
For each experimental group 100 readings were the RC-CS was less than that in the N rats (P < 0.5) and
estimated. Each reading of the 100 readings is an average not different from the RC-CS in the D rats.
of four measurements taken as 2  2 readings along two
perpendicular axes running through the glomerulus. Each Histological data
one of the four readings represents the distance between
the external surface of the glomerulus and the opposite The images of each experimental group were acquired
inner surface of the capsular membrane. The two from different sections of renal tissue representing the

Table 1 Serum glucose concentration and urine protein clearance at the seventh week
N D D þ GC D þ GR
a b
Serum glucose (mg/dl) 129  9 (n Z 10) 431  1 (n Z 10) 237  (n Z 10) 267  2b (n Z 10)
Urine protein (mg/24 h) 7  1 (n Z 10) 13  2a (n Z 10) 6  1b (n Z 10) 5  2b (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.
Effects of garlic and ginger on diabetic nephropathy e65

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 D þ GC still looked larger with some chromatin thinning,
collectively, provided an overall picture of the type of while those of the D þ GR appeared denser.
structural change and status of the renal tissue in each Meticulous examination of the RC at the magnification of
experimental group. 40 (DIC-2 e Fig. 2) revealed that the RC of the D rats
Examination of the digital images combination (DIC-1 e showed early signs of irregular form, considerable shrink-
Fig. 1) of the renal cortices (CX) at low magnification age in the CS to the point where in some places and along
(10) revealed the following: Compared to the CX of the a good length the capsular wall touched the glomerulus.
N rats the CX of D rats had a similar overall architecture. The capsular wall at certain segments lost definition to
However, the majority of the renal corpuscles (RC) in the the point of distortion, whereas at other places some thick-
D rats’ kidneys appeared to have distorted capsular shape ening of the basement membrane was obvious. The glomer-
that reflected on the form of the glomeruli. Furthermore, ulus of the D rats’ kidney showed signs of hypertrophy and
most of the RC-CS in the D rats’ kidney showed the uneven distribution with more condensation of mesangial
presence of protein shedding (capsular drops) that may cells and capillary epithelial cells and in some places fusion
represent insudation of the plasma proteins. Although of mesangial cells. An interesting glomerular structural
these protein markers are evident in the N rats’ capsular change present in the D rats’ kidney was streaks of conden-
space (CS), they were more prevalent in the RC-CS of D sations that radiated through the glomerular bed originat-
rats. The general form of the proximal tubule (PT), distal ing and extending from the vascular pole. In the kidneys
tubule (DT) and collecting duct (CD) appear normal at this of the D þ GC or D þ GR, and compared to the kidney of
magnification. the D rats, the RC showed better corpuscular form (oval
The CX of the D þ GC and D þ GR showed RC with more to round), capsular wall integrity, capsular space dimen-
rounded shape almost similar to that seen in the CX of sions, glomerular layout and nuclear shape. Furthermore,
the N rats. Nonetheless, some structural irregularities are after garlic or ginger treatments there were less condensa-
still evident in the RC of the D þ GC and D þ GR. The PT tion deposits, mesangial fusion, and protein shedding.
and DT general shapes are consistent with those seen in The PT in kidneys of the D rats showed several alterations
the kidney of N rats. The nuclear appearance is different indicating distinct changes compared to the structure of the
between the D þ GC and D þ GR, were the nuclei in the PT in the kidney of N rats. In the D rats’ kidney, the PT
e66 K. Al-Qattan et al.

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 nuclei. The cytoplasm of the PT in the D þ GR seemed
thick in others. The cytoplasm was fragmented and basely intact compared to that seen in the PT of D rats; however
retracted. Most of the nuclei appeared enlarged and demon- it showed signs of granulation. The lumens in the D þ GR
strated chromatin thinning. The tubular lumens showed were larger than those seen in the PT of the D þ GC, but
sizable light globular formations surrounded by dark borders were slightly smaller than those observed in the PT of the
in shades of gray. The peritubular capillary vessels around N and D rats.
the PT appeared enlarged and dilated. The digital image of the upper medullary region (DIC-4
In comparison to the PT of the D rats, the PT of the eFig. 4) suggested that the D þ NS had normal distribution
D þ GC generally had more normal cell distribution and of Henle’s tubules. However, detailed examination of these
architecture. In addition, the PT of the D-GC had thinner tubules reveal that they have slightly thicker basement
basement membranes that nicely supported and made membranes and loss of tubular definition of the cellular
good contact with the cytoplasm of good sized and intact and luminal form, in addition to cytoplasm fragmentation
cuboidal epithelial cells that had well-rounded nuclei. in both the ascending and descending limbs. The nuclei of
The cytoplasm of these cells showed lesser fragmentation, the epithelial cells in the ascending tubules appear as
but exhibited higher intensity of staining. The tubular darkly stained dots that concentrate closer to the center.
lumens were smaller and in some cases, the lumen was The vasa recta vessels are more pronounced with thicker
no more than a radiating center lined by the brush border. basement membranes and larger nuclei compared to those
As is the case with the PT of the three other groups of rats, in the normal vasa recta. Furthermore, the D rats’ vasa
the PT of the D-GR showed a normal gross architecture. The recta borders are rougher in appearance than those noticed
PT of the D þ GR are well defined by a slightly thicker in the N rats.
basement membrane compared to the PT of the N þ NS The upper medulla of the D þ GC kidney showed a typical
and D þ GC, but are slightly thinner than those seen in layout of descending and ascending limbs of the loop and
the PT of the D þ NS. The PT of the D þ GR are internally other structures. Compared to the D rats’ kidney, the limbs
lined with normal size cuboidal epithelial cells that made in the kidney of the D þ GC had better definition of cuboidal
good contact with the basement membrane and surround cells and evenly distributed, slightly denser cytoplasm. The
mostly centrally positioned normal and darkly stained nuclei were well defined and centrally located, but showed
Effects of garlic and ginger on diabetic nephropathy e67

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 Discussion

D þ GC kidney still showed signs of distorted apical cellular
lining. The tubules had better-defined lumens; however, The etiologies of serious and ultimately fatal diseases
less defined basement basal membranes. The upper inherent to diabetes are distinct biochemical and
medulla in the kidney of the D þ GR showed similar struc- physiological abnormalities. It is almost certain that
tural features compared to those seen in the D þ GC upper chronic increases in the concentration of glucose creates
medulla. However, in the tubules of D þ GR there are signs an internal milieu of elevated free radicals that imposes
of better definition of basal membranes and fewer enlarged deleterious stress leading to permanent histological
nuclei. The cuboidal cells in the limbs of the kidney in the damage. This study focused on two of the most established
D þ GR were less defined compared to those observed in the defects that afflict diabetics, namely hyperglycaemia and
D þ GC tubules. structural nephropathy, and investigated the possibility
The digital images of the renal arcuate arteries (RAA) that the hypoglycaemic actions of garlic and ginger might
(DIC-5 e Fig. 5) showed that the artery of the D rats had the attenuate the intensity of structural nephropathy in STZ-
same form as that present in the N rats. However, the RAA induced diabetic rats.
of the D rats showed two distinct structural differences In a review, Ceriello16 stated that hyperglycaemia-
exemplified in the presence of a thicker internal elastic induced overproduction of superoxide seems to be the first
lamina and the presence of more batches of intense and main event in the activation of all pathways involved in
coloration in the tunica media. The thickness of the inter- the pathogenesis of diabetes complications. Furthermore,
nal elastic lamina in the RAA of the D þ GC appears to be increased generation of nitric oxide and, consequently
between those of the N rats and the D rats, whereas, the the formation of the strong oxidant peroxynitrite accompa-
RAA of the D þ GR seems to be as thin as that found in nied superoxide overproduction. In another study,24 it was
the N rats. The tunica media of the RAA in the D þ GC reported that STZ-injected rats displayed variable changes
and D þ GR still showed some intense coloration; nonethe- in homeostatic levels of glucose, total lipids, triglyceride,
less, the intensity of this coloration was slightly less. The cholesterol lipid peroxides, nitric oxide and uric acid.
condition of the endothelium in the RAA of all groups could Concomitantly, significant decreases in the levels of the
not be ascertained at a magnification of 40. antioxidants ceruloplasmin, albumin and total thiols were
e68 K. Al-Qattan et al.

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 form of coronary calcifications.11 It is suggested that most
from the previous reports that the concentrations of power- of these changes result from membranous distortions that
ful oxidative/nitrosative free radicals increase while those could be due to peroxidations of membrane lipids and
of antioxidants decrease in the circulation of diabetics. As damage to membrane protein structure extending from
a result, the amounts of radical agents transported in the the primary to quaternary level.
blood to different tissues and organs of the body become In this study, the terminal values of serum glucose and
pathologically elevated. urine protein of the four groups of rats shown in the table
Both kidneys receive an estimated 20% of the cardiac (and when compared to each other) present a comprehen-
output, which is high considering that their combined sive conclusion regarding the type of changes in the levels
weight does not exceed 0.5% of the total weight of the of these two parameters over the course of the seven
body. In diabetes, as large volumes of blood are delivered weeks of the study. This is supported by detailed periodic
with each heartbeat, the kidneys become perfused with measurements and analysis of serum glucose and urine
large amounts of radical toxins.11 The renal exposure to protein, in addition to the body weight, water intake and
toxins is augmented further by the kidneys’ own production urine output (þ serum levels of triglyceride and
of free radicals which becomes elevated in condition of cholesterol) reported in two recent and related studies in
diabetes. As such, because of the exposure to free radicals the same groups of rats.20,30 In addition to improvements
from both extrinsic and intrinsic sources, the kidneys are in serum glucose level and urine protein clearance in these
regarded as a primary target to the deleterious effects of two studies, the data show that treating diabetic rats with
these metabolites.28,29 The histological observations of either garlic or ginger led to: (a) stabilization in the body
the present study provide support to the destructive nature weight of the treated animals close to the initial weight
of these radicals on the diabetic kidneys. The histological and (b) reduction in water consumption and urine output
images of the diabetic normal saline-treated kidneys show toward normal levels. When the body weight, water
clear signs of structural distortions that developed after consumption and urine output data of the three diabetic
a short period of hyperglycaemia and were obvious even groups are collectively considered, they suggest that the
under the low magnification of light microscopy. Early reported beneficial effects of ginger and garlic are not
changes in tissues structure have also been reported in due to a change in diet. This conclusion is supported by
the diabetic cardiovascular system materializing in the 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 as proteases, which particularly damage and digest the glo-
diabetics) was associated with reduction in both serum merular filtration barrier.32 The deteriorating effects that
glucose and urine protein clearance. the glomerulus experienced is further indicated by these
Most of the structural changes observed in the diabetic observations: (1) the average width of the CS in the dia-
kidneys treated with normal saline were associated with betic RC was significantly reduced in comparison to that
the different segments of the nephron and in particular the measured in the normal RC, which indicates a reduction
components of the RC and associated arterioles. It appears
that the glomerulus is a principal site for the action of
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 K. Al-Qattan et al.

in filtration rate and (2) the daily clearance of protein was This structural cytoplasmic change will diminish the PT
significantly elevated indicating proteinuria, a parameter reabsorptive capacity allowing a considerable quantity of
established as a marker of diabetic nephropathy.33 filtered load of amino acids and proteins to pass into the
It is well known that free radicals by virtue of their urine unclaimed by the diabetic kidneys. This reduction in
ability to extract or donate electrons change the valence PT reabsorptive function may be aggravated by (1) damage
characteristics of affected atoms. This may lead to altering to the absorptive membrane due to loss of microvilli
atomic reactivity, modulating molecular structure, hence observed in this study and reported elsewhere21 and (2)
causing abnormalities in the configuration of major classes destruction of the mitochondria of PT epithelia, which are
of structural biomolecules. It is expected that controlling important for providing the energy necessary for active
the endogenous generation and bioavailability of free transport mechanisms. In addition, it is reported that dam-
radicals will preserve normal atomic physical and chemical aged mitochondria release large amounts of reactive agents
properties and thus reactivity. The view that consumption that expose the diabetic kidneys to more of these deleteri-
of antioxidant-rich foods is important in combating disease ous agents.37 In this study, the injurious effects that the
is receiving acceptance and support.34 Garlic and ginger diabetic kidneys are exposed were further highlighted in
have been shown to possess the potential to effectively the form of cytoplasmic distortion in the upper parts of
ameliorate a considerable number of diabetic biochemical the renal loops located in the upper medullary region.
and physiological modulations targeted in investigative The argument to the cause of this damage is similar to
studies. In addition, the amounts of garlic and ginger used that presented for the PT. The garlic or ginger-treated
in this study were reported to have no damaging effects rats showed lower incidences of change in this segment of
on the structures of tissues such as lung or liver in normal the nephron.
rats.35,36 These corrective actions have been suggested to The vascular endothelium has been reported to synthe-
be partially due to their content, as other plants species, size and release a wide range of vasoactive substances,
of several biologically active compounds that possess including the vasodilator NO, prostacyclin and endothe-
antioxidant properties.34 In the present study, the serum lium-derived hyperpolarizing factors, in addition to the
glucose data showed clearly that garlic and ginger restrain vasoconstrictors angiotensin II and endothelin-1.14 The or-
the level of hyperglycaemia resulting from the experimen- chestrated function of these vasoactive components under
tal destruction of beta pancreatic cells induced by STZ. The normal physiological conditions is imperative to maintain-
hypoglycaemic actions of garlic and ginger appeared to ing normal blood perfusion and circulation. However, these
attenuate tissue injury that occurred in the kidneys of vasoactive agents can alter the circulation following endo-
treated diabetic rats which is suggested to be due to a re- thelial dysfunction that occurs under circumstances such
duction in the level of oxidants.24 The digital images of as trauma, hyperglycaemia or dyslipidaemia.14 Endothelial
the different regions and structures taken from the kidneys dysfunction may result from membranous glycation.38 In
of STZ-injected rats treated with garlic or ginger support the present study, at the level of light microscopy,
the previous claim. The nephrons of kidneys from treated evidence of glycation might be suggested by the distortion
diabetic rats showed a healthier form in particular in the of the endothelial cells of the glomerular vessels, the rough
glomerular region and the surrounding CS. These images appearance of the internal elastic lamina in the RAA, and
suggest lesser intensity of glomerulonephritis than that the rough appearance of the vasa recta in the non-treated
observed in the kidneys of diabetic rats treated with normal diabetic kidneys. Controlling the levels of serum glucose in
saline. The signs of lesser protein shedding observed in the garlic and ginger STZ-injected rats ensured the preserva-
CS of the garlic or ginger-treated rats and the reduction in tion of the structural integrity of the micro- and macro-
the level of proteinuria confirm that the glomeruli in these vessels as suggested by the normal looking glomerular
animals are performing much better than the glomeruli of capillary, elastic lamina of RAA and vasa recta of garlic
control diabetic rats. In support, it was previously reported or ginger-treated kidneys. Thus, in response to these
that garlic and ginger reduce albuminuria, hypertrophy and protective effects of garlic and ginger, it is highly probable
collagen deposition in the glomerulus.8 that the intrinsic humoral regulatory functions of the
The distortion in the PT of the hyperglycaemic rats endothelium of the renal vasculature was maintained as
treated with normal saline suggest further that the supported by the improved protein clearance functions.
increased proteinuria observed in these animals could be It is concluded from the findings of this study that: (1)
due to the impairment of tubular transport mechanisms the distortions in the structure of several nephronal and
dedicated to processing and reclaiming filtered amino acids microvascular components of the kidney materialize in
and proteins. This suggestion is supported by the presence the early stages of diabetes; (2) the degree of renal
of globular forms in the lumens of PT in non-treated STZ structural injury parallels the level of serum glucose; and
rats. Because the PT are a bidirectional venue of reabsorp- (3) garlic or ginger attenuates the progression of struc-
tion and secretion, its cells are susceptible to the injurious tural nephropathy.
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 Acknowledgments
biomolecules, especially proteins, may become damaged
resulting in a distortion of cytoplasmic integrity and form Dr. Khaled Al-Qattan, Dr. Martha Thomson and Professor
in that part of the nephron.5 Cytoplasmic fragmentation, Muslim Ali presented the idea for this investigation,
a main structural feature of diabetic PT pathology,32 has designed the investigation protocol, induced diabetes,
also been observed in this study in the control STZ-kidney. collected and analyzed samples, carried out histological
Effects of garlic and ginger on diabetic nephropathy e71

preparation, examination and capturing of digital images of nephrocalcinosis, and nephrolithiasis. In: Atlas of renal pa-
renal tissue sections, performed statistical analysis, carried thology, vol. 9. W.B. Saunders Company; 1999. p. 97e112.
out literature search and cross-referencing, and wrote the 18. Blumenthal M, Hall T, Goldberg A, et al. The ABC clinical guide
article. to garlic. American Botanical Council; 2003. p. 1e20.
19. Sekiya K, Ohtani A, Kusano S. Enhancement of insulin sensitiv-
We like to thank Mrs. Zainab Al-Amin for assisting in the
ity in adipocytes by ginger. BioFactors 2004;22:153e6.
induction of diabetes and determination of serum glucose 20. Al-Amin ZM, Thomson M, Al-Qattan KK, et al. Anti-diabetic and
and urine protein levels, Ms. Dalal Al-Hajeri for assisting in hypolipidaemic properties of ginger (Zingiber officinale) in
the preparation of tissue sections and measurement of streptozotocin-induced diabetic rats. Brit J Nutr 2006;96:
renal corpuscle capsular space, and Kuwait University for 660e6.
partially supporting this study. 21. Banerjee SK, Maulik M, Manchanda SC, et al. Garlic-induced
alteration in rat liver and kidney morphology and associated
changes in endogenous antioxidants status. Food Chem Toxicol
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