Biomedicine & Pharmacotherapy 95 (2017) 317–323

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Biomedicine & Pharmacotherapy

journal homepage: www.elsevier.com/locate/biopha

Original article

Effect of zinc gluconate, sage oil on inflammatory patterns and MARK

hyperglycemia in zinc deficient diabetic rats
⁎
Mohamed M. Elseweidya, , Abdel-Moniem A. Alib, Nabila Zein Elabidinec, Nada M. Murseyc
a
Department of Biochemistry, Faculty of Pharmacy, Zagazig University, Zagazig 44519, Egypt
b
Department of Pathology, Faculty of Veterinary Medicine, Zagazig University, Zagazig 44519, Egypt
c
Department of Biochemistry, Faculty of Sciences, Zagazig University, Zagazig 44519, Egypt

A R T I C L E I N F O A B S T R A C T

Keywords: Background: The relationship between zinc homeostasis and pancreatic function had been established. In this
Type1 diabetes mellitus study we aimed firstly to configure the inflammatory pattern and hyperglycemia in zinc deficient diabetic rats.
Inflammatory markers Secondly to illustrate the effect of two selected agents namely Zinc gluconate and sage oil (Salvia Officinalis,
Zinc deficiency family Lamiaceae).
Salvia officinales
Methods: Rats were fed on Zinc deficient diet, deionized water for 28 days along with Zinc level check up at
Zinc gluconate
intervals to achieve zinc deficient state then rats were rendered diabetic through receiving one dose of alloxan
Zinc transporter 8 (Znt8)
monohydrate (120 mg/kg) body weight, classified later into 5 subgroups.
Results: Treatment with sage oil (0.042 mg/kg IP) and Zinc gluconate orally (150 mg/kg) body weight daily for
8 weeks significantly reduced serum glucose, C-reactive protein (CRP), Tumor necrosis factor alpha (TNF- α),
interleukins-6 1 β, inflammatory8 (IFN ȣ), pancreatic 1L1-β along with an increase in serum Zinc and pancreatic
Zinc transporter 8 (ZNT8). Histopathological results of pancreatic tissues showed a good correlation with the
biochemical findings.
Conclusions: Both sage oil and zinc gluconate induced an improvement in the glycemic and inflammatory states.
This may be of value like the therapeutic agent for diabetes.

1. Introduction
Type one diabetes mellitus (T1DM) is a complex autoimmune dis-
ease, mostly attributed to β-cells destructions by auto reactive T-cells
leading to severe insulin deficiency [1].
The early Diagnosis of diabetic individuals usually demonstrates
higher level of serum inflammatory biomarkers. This indicates that the
inflammatory response is activated during the early stages of the dis-
ease [2]. Zinc is one of the most important trace elements in biological
systems [3]. It is found in plasma bound to albumin and α- macro-
globulin, achieving less than 1% of the Zinc total body contents while
the remaining (99%) are located intracellularly [4].
Zinc protects biological structures against free radical inducing
damage through the maintenance of metallothioneins. The latter is an
essential component of superoxide dismutase enzyme (SOD) which
protects against oxidative stress [5].
Zinc also plays an important role in pancreatic islets as a specific
structural component of the insulin molecule(Zinc- insulin) complex
and also in insulin secretion [6]. It is involved in many processes within
the islets like glucagon secretion, insulin packaging, signaling and se-
cretion as well as the digestive enzyme activity [6].
Therefore, deregulation of Zinc metabolism may impair many key
processes including glycemic control [7], pancreatic cancer [8,9] and
chronic pancreatitis [10].
It is clear now that Zinc is co-secreted with insulin into the islet
extracellular space, its release from insulin usually occurs at higher pH
of the blood. Furthermore, Zinc ions provide an off-switch for glucagon
release from the α-cells during glucose deprivation through closure of
α-cell KATP channel [11,12].
Zinc is actually transported into pancreatic β-cells via certain
transporters, ZnT8 is an example of a protein which transports Zinc into
insulin granules. Release of auto antibodies to ZnT8 and its poly-
morphism is associated with the onset of Diabetes [13].
The Genus Salvia and its different species are commonly known as
sage. Salvia Officinalis species which is referred to as common sage is
native to the Mediterranean region and has been used as flavoring
species as well as traditional herbal medicine [14]. Sage oil has been
reported to have many biological activities such as antioxidant,

⁎
Corresponding author
E-mail addresses: mmelseweidy@yahoo.com (M.M. Elseweidy), Abdelmoniemali@yahoo.com (A.-M.A. Ali), Nabila.zein@yahoo.com (N.Z. Elabidine),
kateralnada@gmail.com (N.M. Mursey).

http://dx.doi.org/10.1016/j.biopha.2017.08.081
Received 4 August 2017; Received in revised form 18 August 2017; Accepted 21 August 2017
0753-3322/ © 2017 Elsevier Masson SAS. All rights reserved.
M.M. Elseweidy et al.
antibacterial [15], hypoglycemic [16] additionally anti-inflammatory
properties [17].
Therefore, the present study aims to first illustrate the profile of Zinc
deficient state in alloxan-diabetic-rats put on Zinc deficient diet.
Secondly, the effect of sage oil and zinc gluconate supplementation
individually on certain inflammatory and Zinc transporter biomarkers
of the present animal model.
This may demonstrate their potentials as effective antidiabetic,
antioxidant and anti-inflammatory properties.
2. Materials and methods
Thirty adult male albino rats weighing 170 ± 20 g were housed
under environmentally-controlled conditions and were allowed one
week acclimatization at room temperature with a 12 h’ dark/light cycle
before beginning the experimental work. Rats were fed rodent chow
and allowed free access of drinking water. The animals were main-
tained, used in accordance with the Animal Welfare Act, the Guide for
the Care and Use of Laboratory Animals (The University of Zagazig,
Egypt). Rats were fed on zinc deficient diet for 28 days and consist of:
Diet contents Quantity (g/kg)
Egg albumin 200
Dextrin 631
Maiz oil 100
Vitamin mixture 11.7
Salt mixture (free of zinc) 31.3
Cellulose powder 20
Deionized water was allowed as drinking water for all rats [18].
Single dose (120 mg/kg) of freshly prepared solution of Alloxan
Monohydrate which was purchased from the ACROS; ORGANIC-
CHEMICALS COMPANY (New Jersey, USA)(Dissolved in Normal Saline,
Citrate buffer, pH 4.5) was administered intraperitoneal to overnight
fasting rats for the induction of diabetes mellitus. Control rats were
similarly injected with normal saline. Fasting blood glucose level was
checked after 48–72 h. Rats which achieved a blood sugar level >
200 mg/dl were selected as diabetics [19]. Seven experimental groups
(n = 6) were used: normal rats (NC) which received no treatment and
served as Normal group, Diabetic control,normal rats and received al-
loxan only (DC), Zinc deficient group (znd) which received zinc defi-
cient diet and deionized Water only and diabetic zinc deficient group
(Dz.d) which received alloxan and kept on Zinc deficient diet and
deionized water. The last group was subclassified into Three groups.
The first one (S.O group) diabetic zinc deficient rats, injected by sage
essential oil (0.042 mg/kg body weight) intraperitonially [20], daily for
8 weeks. The second one received Zinc gluconate. (Z.G group) 150 mg/
serving water for 8 weeks daily [21] while the third one received no
drugs and referred to as Diabetic rats kept on Zinc deficient diet and
deionized water and referred to as control for comparison with the
drugs used. During the experimental period (8 weeks), body weight,
blood glucose, food and water consumption and physical examinations
were determined at regular intervals. The dosage was adjusted weekly
in response to any changes in body weight.
2.1. Blood sampling
At the end of the treatment periods (8 weeks) rats were fasted
overnight, blood samples were taken for each rat individually and di-
rected to serum preparation. Samples were processed instantly for de-
termination of glucose, IL-6, CRP, and TNF-α, IFN-γ, IL1-β and Zn.
318
Biomedicine & Pharmacotherapy 95 (2017) 317–323
2.2. Tissue collection
After blood collection, rats were killed by decapitation, pancreatic
tissues were removed instantly, placed in cold saline solution, trimmed
of adipose tissue and homogenized instantly on ice using buffer. Tissue
homogenate stored at −20 °C for determination of pancreatic IL1-β and
ZNT8.
2.3. Histopathological examination of pancreatic tissues
A slice of pancreas was fixed in 10% formalin for 1week at room
temperature, the specimens were dehydrated in a graded series of
ethanol cleared in xylene, and embedded in paraffin wax Tissue blocks
were sectioned to 4-μm thick using a rotary microtome. Sections were
stained by hematoxylin and eosin. Stained sections were examined by
light microscope [22].
2.4. Analytical methods
Blood glucose determination was done using commercial kits sup-
plied by Spinreact Kits, Barcelona, Spain [23]. Serum IL-1beta was
determined using commercialELISA kit supplied by R & D quantikine
(USA) [24]. C-reactive protein (CRP) wasdetermined using BD Bios-
ciences ELISA Kit, USA [25]. Serum IL-6 was determined using com-
mercial ELISA kitsupplied by R & D quantikine (USA) [26]. Serum TNF-
α was determined according to the method of Juhasz, 2013 [27] using
R & D Quantikine ELISA Kit (USA). Serum INFγ was determined ac-
cording to the method of [28] using Platinum ELISA Kit (USA). Serum
ZN was determined by colorimetric method according to [29] using
QCA –QuímicaClínicaAplicada S.A., Spain. Znt-8 was determined using
AnticropsELISA Kit (Aachen, Germany).
2.5. Statistical analysis
All results were expressed as Means ± SD. Statistical analysis and
correlation were performed using Graph Pad Prism software. Student
“t” test of unpaired data and the analysis of variance (one-way ANOVA)
followed by Tukey’s post hoc test were used for comparison between
groups. The correlations between the studied parameters were done
using the Pearson’s correlation coefficient (r). Statistical significance
was defined at P < 0.05 [30,31].
3. Results
3.1. Biochemical parameters
Zinc deficient (Znd), diabetic control (DC) and diabetic zinc defi-
cient group (Dz.d) demonstrated significant increase in serum glucose,
IL-6, CRP, and TNF-α, IFN-γ, IL1-β and in pancreatic IL1-β along with
significant decrease in serum zinc and pancreatic ZNT8 content in
comparison to normal group (Table 1). Dz.d group demonstrated also
significant increase in serum glucose, IL-6, CRP, and TNF-α, IFN-γ, IL1-
β and in pancreatic IL1-β along with significant decrease in serum zinc
and pancreatic ZNT8 in comparison to diabetic group (DC) (Table 1).
Treatment with S.O (sage oil) and zinc gluconate for 8 weeks resulted in
significant decrease of serum glucose and the inflammatory markers as
compared to the Dz.d group (control group, Table 2), The sage oil po-
tential showed characteristic pattern than zinc gluconate as hypogly-
cemic and anti-inflammatory agent. Histopathological findings de-
monstratedgood correlation with the Biomarkers results (Fig. 2).
3.2. Zinc modulation parameters
Serum zinc and pancreatic ZNT8 showed Negative correlation with
serum glucose (Fig. 1a, Fig. 1b) but positively correlated with each
other (Fig. 1c).
M.M. Elseweidy et al. Biomedicine & Pharmacotherapy 95 (2017) 317–323

Table 1
Serum and pancreatic Biomarkers of diabetic and Zinc deficient diabetic group of Experimental rats after 8 weeks as compared to normal group.

Parameter NC DC Zd Dz.d

Serum glucose (mg/dl) 79.5 ± 6.8 487.7 ± 41.9 ** 517 ± 35** 547.0 ± 30.2**##
Serum IL-6 (pg/ml) 32.9 ± 2.2 48.5 ± 4.2** 75 ± 6.5 ** 103.6 ± 9.5**##
Serum TNF-α (pg/ml) 34.7 ± 2.5 118.8 ± 7.8** 121 ± 7.5** 124.3 ± 8.2 **
Serum CRP (pg/ml) 1.02 ± 0.04 12.1 ± 1.2** 12.4 ± 1.0** 12.7 ± 0.8**
Serum IFN-γ (pg/ml) 0.97 ± 0.04 11.1 ± 1.1** 11.7 ± 1.4** 12.2 ± 0.7 **#
Serum IL–1 B (pg/ml) 40.2 ± 3.5 120.5 ± 7.3** 120 ± 7** 128.6 ± 7.6**
Serum ZN (ug/ml) 1.06 ± 0.08 0.19 ± 0.06** 0.02 ± 0. 001.** 0.02 ± 0.0006**##
Pancreatic ZNT8 (ug/Gl) 1.002 ± 0.05 0.69 ± 0.06** 0.25 ± 0.05** 0.27 ± 0.03**##
Pancreatic IL 1-B (pg/Gl) 39.37 ± 4.36 116.4 ± 8.6** 120 ± 5.1** 24.4 ± 4.98**

NC, normal group, Dc: diabetic control group, Zd: Zn deficient group, Dz.d: diabetic zinc deficient group, n = 6 in each case. Values are expressed as mean ± S.D. significant differences
are shown:*p < 0.05, ** p < 0.001 vs. NC group, # p < 0.05,# # p < 0.001 vs. DC group.

Table 2 Regenerative attempts – – – 1+ 1+

Serum and pancreatic biomarkers of Diabetic Zinc deficient rats received Sage oil (SO)
and Zinc gluconate individually for 8 weeks as compared to control group. (− = negative changes. 1+ = Mild. 2+ = Modate.
3+ = Severe).
Parameter Control Gp (DZd) SO group Zn Gluconate group

Serum glucose (mg/ 547 ± 30.2 94.17 ± 6.8# 120.2 ± 7.41# 4. Discussion
dl)
Serum IL-6 (pg/ml) 103.6 ± 9.5 30.73 ± 0.86# 57.4 ± 5.19#
In agreement with reported studies [32–34] the results demon-
Serum TNF-α (pg/ 124.3 ± 8.2 35.65 ± 2.9# 65.5 ± 4.77#
ml) strated that alloxan administration significantly increased serum and
Serum CRP (pg/ml) 12.7 ± 0.8 0.9 ± 0.07# 5.33 ± 0.42# pancreatic inflammatory markers, e.g. IL-6, CRP, TNF-α, IL-1β, IFN-ϒ
Serum IFN-γ (pg/ 12.2 ± 0.7 6.72 ± 0.06# 4 ± 0.35# as compared to the normal group. This increase in inflammatory cyto-
ml) kines can stimulate the hepatocytes to increase the synthesis and re-
Serum IL–1 B (pg/ 128.6 ± 7.6 41.03 ± 1.2# 74.3 ± 6.45#
ml)
lease the positive acute-phase proteins along with the activation of the
Serum ZN (ug/ml) 0.02 ± 0.006 0.95 ± 0.05# 1.83 ± 0.23# innate immune activity [35,36].
Pancreatic ZNT8 0.27 ± 0.03 1.1 ± 0.1# 1.13 ± 0.07# Both Serum Zinc and pancreatic ZnT8 levels of diabetic rats showed
(ug/Gl) also significant decrease as compared to the normal group and both
Pancreatic IL 1-B 124.4 ± 4.98 39.66 ± 2.2# 78.5 ± 4.56#
levels are negatively correlated with serum glucose (P < 0.0001). In
(pg/Gl)
general diabetics of both types usually achieve decreased serum Zinc
Dzd (Diabetic Zn deficient group), SO (sage group), Zn Gluconate group. values are ex- level [37–39] along with increased loss of urine zinc [40]. This may
pressed as M ± SD. significance differences are shown # P ≤0.05 vs. Dzd group (con- refer to Zinc deficiency as a consequence of hyperglycemia or mostly
trol). contributes to the pathogenesis of diabetes.
Reduced function of ZnT8 may decrease the Zinc content of the islet
3.3. Histological changes cells additionally reduce its secretion [41]. This in turn may impair the
β-cell function leading to an increase of oxidative stress [42,43]. This is
Endocrine and exocrine pancreas of normal control rats appeared logic since dietary zinc restriction induces peripheral and central al-
within the normal histomorphological pictures (Fig. 2A, B). Alloxan- terations of bio-elements (namely zinc and iron), enhances oxidative
diabetic endocrine pancreas revealed reductions in both Islets size and damage and pro-inflammatory status [43–45].
number together with cell vaculation or deaths (apoptosis) and deple- Zinc alteration may also affect peripheral glucose metabolism
tion of cell mass (Fig. 2C). The for mentioned lesions were encountered through its effect on translocation of the glucose transporters inside the
in diabetic-zinc deficient group in addition to amyloid deposits within cells or its structure modification where deficiency of zinc can decrease
the matrix of some Islets and intense lymphocytic infiltrations (Fig. 2D). tissue response to insulin [46].
Regarding to Salvia oil and Zinc gluconate treated groups, the majority Reported studies indicated that Zinc deficient patients demonstrate
of pancreatic Islets restore their sizes, number and cell population be- also significant increase in reactive oxygen species (ROS) and pro-in-
side some regenerative attempts; meanwhile a few cells still vaculated flammatory cytokines [45,47,48].
and other hyperplastic and hypertrophied (Fig. 2E, F). A previous in vitro studies indicated that Increased glucose con-
Microscopic lesion score of pancreas among different experimental centration alone has been suggested to induce apoptosis in human β-
groups. cells through the induction of IL-1β, leading to NF-kB activation and up-
regulation of Fas receptors [49,50]. Therefore, anti-oxidant effect can
confer significant protection against the oxidative stress and damage
Criteria Control Diabetic DZ.d DZ.d DZ.d induced by free radicals.
+S +Z. In agreement with reported clinical and experimental studies
oil G [41,51,52], Zinc intake in the present study significantly decreased
blood glucose level which may be attributed to blocking of the NF-kβ
Reduction in Islets sizes – 3+ 3+ 1+ 1+
activation in pancreatic tissues since the latter is sensitive to ROS [53].
and numbers
The potential mechanism for the insulin-like effects of Zinc ions is due
Islets cell Vacoulations – 2+ 2+ 1+ 1+
to an enhancement of tyrosine phosphorylation of multiple receptor
Matrix amyloid deposits – 1+ 2+ – –
protein tyrosine kinases (R-PTKS). The latter include insulin receptor
Islets cell deaths – 3+ 3+ 1+ 1+
(IR), insulin –like growth factor type-1 receptor (IGF-IR) and Epidermal
(apoptosis) and
growth factor receptor (EGFR) [54]. This is achieved through the pro-
depletions of cell mass
duction increase of ROS species and subsequent inhibition of protein
Lymphocytic infiltration – – 2+ – –
tyrosine phosphatase (PTPase), leading to an increase in tyrosine

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M.M. Elseweidy et al. Biomedicine & Pharmacotherapy 95 (2017) 317–323

Fig. 1. (a) Negative correlation of pancreatic zinc transporter 8 with serum glucose, (b) Negative correlation of serum zinc with serum glucose, (c) Positive correlation of pancreatic zinc
transporter 8 with serum zinc.

phosphorylation of R-PTK [54]. The latter phosphorylates multiple possible future therapeutic candidate for diabetic treatment. Further
downstream targets, inducing the phosphorylation and activation of the clinical studies are recommended in the future to confirm the experi-
MAPK/ERK1/2 and the P13-k/PKB/AKT signaling pathways finally mental results.
leading to an increase of glucose uptake, its transport and glycogen
synthesis [54].
Ethics approval
Previous studies indicated that Zinc supplementation improved Th1
cell cytokines production and decreased the generation of inflammatory
The study has been approved from ethics committee of Faculty of
cytokines [43,55]. Additional studies referred to the potential of Zinc to
Pharmacy, Zagazig University and the animals were maintained and
increase insulin sensitivity, decrease oxidative stress additionally in-
used in accordance with the Animal Welfare Act and the Guide for the
flammation status [56–59]. Present results confirm that Zinc gluconate
Care and Use of Laboratory Animals (The University of Zagazig, Egypt).
intake has increased serum zinc level and pancreatic ZnT8 content.
Sage oil intake significantly decreased the blood glucose level of the
diabetic rats and in line with previous studies which demonstrated its Consent for publication
traditional remedy as hypoglycemic agent, in additional to these in-
vestigations that were done using experimental animals [60,61]. Hy- Not applicable.
poglycemic effect of sage oil may be attributed to its antioxidant ac-
tivity and its high content of polyphenol compounds acting as
scavenger for free radicals DPPH and ABTS [62–64]. Pathological re- Availability of data and material
sults showed to certain extent a good correlation with the biochemical
findings. Please contact the authors for data requests.

Funding
5. Conclusion
There was no funding from any organization or anybody for this
The individual administration of sage oil and zinc gluconate re-
study.
sulted in a marked improvement in the glycemic state associated with
its inflammatory pattern in diabetic rats as compared to the control
group. Histopathological examination demonstrated certain attenua- Competing interests
tion to pancreatic islets and acinar cells induced damage and offered
great support to the biochemical findings. This may be of value as The authors declare that they have no competing interests.

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M.M. Elseweidy et al. Biomedicine & Pharmacotherapy 95 (2017) 317–323

Fig. 2. Photomicrographs of pancreas of rats stained with H & E showing normal histomorphological pictures of both endocrine and exocrine pancreas (A, B). Reduction in Islets size and
cell mass of diabetic group (C). Amyloid deposits (star) and lymphocytic infiltration (arrow) of diabetic-zinc deficient diet (D). Regenerative attempts with hyperplasia of Islet cells
(arrow) in pancreas of treated groups (E, F).

Authors’ contributions References

MM designed the work and the idea of the study and revised the [1] F. Gorus, I. Weets, P. Couck, D. Pipeleers, Epidemiology of type 1 and type 2 dia-
manuscript. betes. The added value of diabetes registries for conducting clinical studies: the
Belgian paradigm, Acta Clin. Belg. 59 (1) (2004) 1–13.
AA carried out the histopathology work. [2] K.I. Alexandraki, C. Piperi, P.D. Ziakas, N.V. Apostolopoulos, K. Makrilakis,
NZ carried out the statistical analyses and drafted the manuscript. V. Syriou, E. Diamanti-Kandarakis, G. Kaltsas, A. Kalofoutis, Cytokine secretion in
NM carried out the experimental work. long-standing diabetes mellitus type 1 and 2: associations with low-grade systemic
inflammation, J. Clin. Immunol. 28 (4) (2008) 314–321.
[3] A. Sharma, B. Patni, D. Shankhdhar, S. Shankhdhar, Evaluation of different PGPR
strains for yield enhancement and higher Zn content in different genotypes of rice
Acknowledgements (Oryza sativa L.), J. Plant Nutr 38 (3) (2015) 456–472.
[4] B.L. Vallee, K.H. Falchuk, The biochemical basis of zinc physiology, Physiol. Rev. 73
(1) (1993) 79–118.
We acknowledge the effort presented by Dr. Samieh Eldahmy, [5] M. Stefanidou, C. Maravelias, A. Dona, C. Spiliopoulou, Zinc: a multipurpose trace
professor of phytochemistry during the preparation, purification and element, Arch. Toxicol. 80 (1) (2006) p. 1.
[6] M. Ynsa, M. Ren, R. Rajendran, J. Sidhapuriwala, J. Van Kan, M. Bhatia, F. Watt,
identification of Nigella sativa oil.
Zinc mapping and density imaging of rabbit pancreas endocrine tissue sections
using nuclear microscopy, Microsc. Microanal. 15 (04) (2009) 345–352.
[7] M.-Y. Jou, A.F. Philipps, B. Lönnerdal, Maternal zinc deficiency in rats affects

321
M.M. Elseweidy et al.
growth and glucose metabolism in the offspring by inducing insulin resistance
postnatally, J. Nutr. 140 (9) (2010) 1621–1627.
[8] M. Li, Y. Zhang, U. Bharadwaj, Q.J. Zhai, C.H. Ahern, W.E. Fisher, F.C. Brunicardi,
C.D. Logsdon, C. Chen, Q. Yao, Down-regulation of ZIP4 by RNA interference in-
hibits pancreatic cancer growth and increases the survival of nude mice with
pancreatic cancer xenografts, Clin. Cancer Res. 15 (19) (2009) 5993–6001.
[9] A.K. Jayaraman, S. Jayaraman, Increased level of exogenous zinc induces cyto-
toxicity and up-regulates the expression of the ZnT-1 zinc transporter gene in
pancreatic cancer cells, J. Nutr. Biochem. 22 (1) (2011) 79–88.
[10] H. Milnerowicz, M. Jablonowska, A. Bizoń, Change of zn, cu and metallothionein
conc and the cu-zn superoxide dismutase activity in patients with pancreatitis,
Pancreas 38 (2009) 681–688.
[11] M. Slucca, J.S. Harmon, E.A. Oseid, J. Bryan, R.P. Robertson, ATP-sensitive K+
channel mediates the zinc switch-off signal for glucagon response during glucose
deprivation, Diabetes 59 (1) (2010) 128–134.
[12] A. Hardy, A. Serino, N. Wijesekara, F. Chimienti, M. Wheeler, Regulation of glu-
cagon secretion by zinc: lessons from the βcell-specific Znt8 knockout mouse model,
Diabetes Obes. Metab. 13 (s1) (2011) 112–117.
[13] S.L. Kelleher, N.H. McCormick, V. Velasquez, V. Lopez, Zinc in specialized secretory
tissues: roles in the pancreas, prostate, and mammary gland, Adv. Nutr.: Int. Rev. J.
2 (2) (2011) 101–111.
[14] D.J. Eide, Zinc transporters and the cellular trafficking of zinc, Biochim. Biophys.
Acta (BBA)-Mol. Cell Res. 1763 (7) (2006) 711–722.
[15] J. Hohmann, I. Zupkó, D. Rédei, M. Csányi, G. Falkay, I. Máthé, G. Janicsák,
Protective effects of the aerial parts of Salvia officinalis, Melissa officinalis and
Lavandula angustifolia and their constituents against enzyme-dependent and en-
zyme-independent lipid peroxidation, Planta Med. 65 (06) (1999) 576–578.
[16] F. Alarcon-Aguilar, R. Roman-Ramos, J. Flores-Saenz, F. Aguirre-Garcia,
Investigation on the hypoglycaemic effects of extracts of four Mexican medicinal
plants in normal and Alloxan-diabetic mice, Phytother. Res. 16 (4) (2002) 383–386.
[17] D. Baricevic, S. Sosa, R. Della Loggia, A. Tubaro, B. Simonovska, A. Krasna,
A. Zupancic, Topical anti-inflammatory activity of Salvia officinalis L. leaves: the
relevance of ursolic acid, J. Ethnopharmacol. 75 (2) (2001) 125–132.
[18] N. Tamaki, S. Fujimoto-Sakata, M. Kikugawa, M. Kaneko, S. Onosaka, T. Takagi,
Analysis of cyclic feed intake in rats fed on a zinc-deficient diet and the level of
dihydropyrimidinase (EC 3.5.2.2), Br. J. Nutr. 73 (5) (1995) 711–722.
[19] A. Rohilla, S. Ali, Alloxan induced diabetes: mechanisms and effects, Int. J. Res.
Pharma Biomed. Sci. 3 (2012) 819–821.
[20] M. Eidi, A. Eidi, H. Zamanizadeh, Effect of Salvia officinalis L. leaves on serum
glucose and insulin in healthy and streptozotocin-induced diabetic rats, J.
Ethnopharmacol. 100 (3) (2005) 310–313.
[21] K. Hamden, M.A. Boujbiha, H. Masmoudi, F.M. Ayadi, K. Jamoussi, A. Elfeki,
Combined vitamins (C and E) and insulin improve oxidative stress and pancreatic
and hepatic injury in alloxan diabetic rats, Biomed. Pharmacother. = Biomed.
Pharmacother. 63 (2) (2009) 95–99.
[22] J.D. Bancroft, M. Gamble, Theory and Practice of Histological Techniques, Elsevier
Health Sciences, 2008.
[23] P. Trinder, Determination of blood glucose using 4-amino phenazone as oxygen
acceptor, J. Clin. Pathol. 22 (2) (1969) 246.
[24] J.E. Sims, D.E. Smith, The IL-1 family: regulators of immunity, Nat. Rev. Immunol.
10 (2) (2010) 89–102.
[25] N.D. Padilla, W.K. Bleeker, Y. Lubbers, G.M. Rigter, G.J. Van Mierlo, M.R. Daha,
C.E. Hack, Rat C-reactive protein activates the autologous complement system,
Immunology 109 (4) (2003) 564–571.
[26] M. Peters, A.M. Muller, S. Rose-John, Interleukin-6 and soluble interleukin-6 re-
ceptor: direct stimulation of gp130 and hematopoiesis, Blood 92 (10) (1998)
3495–3504.
[27] K. Juhasz, K. Buzas, E. Duda, Importance of reverse signaling of the TNF super-
family in immune regulation, Expert Rev. Clin. Immunol. 9 (4) (2013) 335–348.
[28] Y. Aoki, O. Katoh, Y. Nakanishi, S. Kuroki, H. Yamada, A comparison study of IFN-γ,
ADA, and CA125 as the diagnostic parameters in tuberculous pleuritis, Respir. Med.
88 (2) (1994) 139–143.
[29] T. Makino, M. Saito, D. Horiguchi, K. Kina, A highly sensitive colorimetric de-
termination of serum zinc using water-soluble pyridylazo dye, Clin. Chim. Acta Int.
J. Clin. Chem. 120 (1) (1982) 127–135.
[30] J. Karbowska, Z. Kochan, Fat-reducing effects of dehydroepiandrosterone involve
upregulation of ATGL and HSL expression, and stimulation of lipolysis in adipose
tissue, Steroids 77 (13) (2012) 1359–1365.
[31] K.H. Alzoubi, K.K. Abdul-Razzak, O.F. Khabour, G.M. Al-Tuweiq, M.A. Alzubi,
K.A. Alkadhi, Adverse effect of combination of chronic psychosocial stress and high
fat diet on hippocampus-dependent memory in rats, Behav. Brain Res. 204 (1)
(2009) 117–123.
[32] J.K. Snell-Bergeon, N.A. West, E.J. Mayer-Davis, A.D. Liese, S.M. Marcovina,
R.B. D'Agostino Jr., R.F. Hamman, D. Dabelea, Inflammatory markers are increased
in youth with type 1 diabetes: the SEARCH case-control study, J. Clin. Endocrinol.
Metab. 95 (6) (2010) 2868–2876.
[33] J.S. Reis, C.A.V. Amaral, C.M.O. Volpe, J.S. Fernandes, E.A. Borges, C.A. Isoni,
P.M.F.d. Anjos, J.A.N. Machado, Oxidative stress and interleukin-6 secretion during
the progression of type 1 diabetes, Arquivos Bras. Endocrinol. Metab. 56 (7) (2012)
441–448.
[34] A.M. Saud, Serum levels of tumor necrosis factor alpha and interleukin-12 in some
Iraqi diabetic patients type I, Int. J. Curr. Microbiol. Appl. Sci. 3 (4) (2014)
260–268.
[35] A. Zaciragic, N. Mulabegovic, J. Huskic, A. Valjevac, N. Avdagic, A. Hadzovic-
Dzuvo, O. Lepara, S. Dobraca, S. Dinarevic-Mesihovic, Increased serum C-reactive
protein concentration is associated with body mass index but not with glycated
322
Biomedicine & Pharmacotherapy 95 (2017) 317–323
haemoglobin in patients with type 1 diabetes mellitus with body mass index within
normal range, Br. J. Diabetes Vasc. Dis. 11 (5) (2011) 249–252.
[36] J.G. Bode, U. Albrecht, D. Häussinger, P.C. Heinrich, F. Schaper, Hepatic acute
phase proteins–regulation by IL-6-and IL-1-type cytokines involving STAT3 and its
crosstalk with NF-κB-dependent signaling, Eur. J. Cell Biol. 91 (6) (2012) 496–505.
[37] A.E. Long, A.T. Gooneratne, S. Rokni, A.J. Williams, P.J. Bingley, The role of au-
toantibodies to zinc transporter 8 in prediction of type 1 diabetes in relatives: les-
sons from the European nicotinamide diabetes intervention trial (ENDIT) cohort, J.
Clin. Endocrinol. Metab. 97 (2) (2011) 632–637.
[38] P. Gunasekara, M. Hettiarachchi, C. Liyanage, S. Lekamwasam, Blood Sugar low-
ering effect of zinc and multi vitamin/mineral supplementation is dependent on
initial fasting blood glucose, J. Diabetol. 1 (2) (2011).
[39] V. Garg, R. Gupta, R. Goyal, Hypozincemia in diabetes mellitus, J. Assoc. Phys.
India 42 (9) (1994) 720–721.
[40] M. Basaki, M. Saeb, S. Nazifi, H. Shamsaei, Zinc, copper, iron, and chromium
concentrations in young patients with type 2 diabetes mellitus, Biol. Trace Elem.
Res. 148 (2) (2012) 161–164.
[41] J. Jansen, E. Rosenkranz, S. Overbeck, S. Warmuth, E. Mocchegiani, R. Giacconi,
R. Weiskirchen, W. Karges, L. Rink, Disturbed zinc homeostasis in diabetic patients
by in vitro and in vivo analysis of insulinomimetic activity of zinc, J. Nutr. Biochem.
23 (11) (2012) 1458–1466.
[42] A. El-Yazigi, N. Hannan, D.A. Raines, Effect of diabetic state and related disorders
on the urinary excretion of magnesium and zinc in patients, Diabetes Res.
(Edinburgh, Scotland) 22 (2) (1992) 67–75.
[43] R. Wegmüller, F. Tay, C. Zeder, M. Brnić, R.F. Hurrell, Zinc absorption by young
adults from supplemental zinc citrate is comparable with that from zinc gluconate
and higher than from zinc oxide, J. Nutr. 144 (2) (2014) 132–136.
[44] M. El Muayed, L.K. Billings, M.R. Raja, X. Zhang, P.J. Park, M.V. Newman,
D.B. Kaufman, T.V. O'Halloran, W.L. Lowe, Acute cytokine-mediated down-
regulation of the zinc transporter ZnT8 alters pancreatic β-cell function, J.
Endocrinol. 206 (2) (2010) 159–169.
[45] A.S. Prasad, Impact of the discovery of human zinc deficiency on health, J. Trace
Elem. Med. Biol. 28 (4) (2014) 357–363.
[46] U. Doboszewska, B. Szewczyk, M. Sowa-Kućma, K. Noworyta-Sokołowska,
P. Misztak, J. Gołębiowska, K. Młyniec, B. Ostachowicz, M. Krośniak,
A. Wojtanowska-Krośniak, K. Gołembiowska, M. Lankosz, W. Piekoszewski,
G. Nowak, Alterations of bio-elements: oxidative, and inflammatory status in the
zinc deficiency model in rats, Neurotox. Res. 29 (1) (2016) 143–154.
[47] L.S. Mayer, P. Uciechowski, S. Meyer, T. Schwerdtle, L. Rink, H. Haase, Differential
impact of zinc deficiency on phagocytosis: oxidative burst, and production of pro-
inflammatory cytokines by human monocytes, Metallomics: Integr. Biometal Sci. 6
(7) (2014) 1288–1295.
[48] R. Jayawardena, P. Ranasinghe, P. Galappatthy, R. Malkanthi, G. Constantine,
P. Katulanda, Effects of zinc supplementation on diabetes mellitus: a systematic
review and meta-analysis, Diabetol. Metab. Syndrome 4 (1) (2012) 13.
[49] C.P. Wong, N.A. Rinaldi, E. Ho, Zinc deficiency enhanced inflammatory response by
increasing immune cell activation and inducing IL6 promoter demethylation, Mol.
Nutr. Food Res. 59 (5) (2015) 991–999.
[50] I. Wessels, H. Haase, G. Engelhardt, L. Rink, P. Uciechowski, Zinc deficiency induces
production of the proinflammatory cytokines IL-1β and TNFα in promyeloid cells
via epigenetic and redox-dependent mechanisms, J. Nutr. Biochem. 24 (1) (2013)
289–297.
[51] C.-C. Lin, H.-H. Huang, C.-W. Hu, B.-H. Chen, I.-W. Chong, Y.-Y. Chao, Y.-L. Huang,
Trace elements: oxidative stress and glycemic control in young people with type 1
diabetes mellitus, J. Trace Elem. Med. Biol. 28 (1) (2014) 18–22.
[52] K. Maedler, P. Sergeev, F. Ris, J. Oberholzer, H.I. Joller-Jemelka, G.A. Spinas,
N. Kaiser, P.A. Halban, M.Y. Donath, Glucose-induced β cell production of IL-1β
contributes to glucotoxicity in human pancreatic islets, J. Clin. Invest. 110 (110 (6))
(2002) 851–860.
[53] P. Gunasekara, M. Hettiarachchi, C. Liyanage, S. Lekamwasam, Effects of zinc and
multimineral vitamin supplementation on glycemic and lipid control in adult dia-
betes, Diabetes Metab. Syndrome Obes.: Targets Ther. 4 (2011) 53–60.
[54] A. Zampelas, Zinc supplementation: another myth or we are heading towards a new
era in the treatment of diabetes? Atherosclerosis 219 (1) (2011) 22–23.
[55] B. Bao, A.S. Prasad, F.W. Beck, J.T. Fitzgerald, D. Snell, G.W. Bao, T. Singh,
L.J. Cardozo, Zinc decreases C-reactive protein, lipid peroxidation, and in-
flammatory cytokines in elderly subjects: a potential implication of zinc as an
atheroprotective agent, Am. J. Clin. Nutr. 91 (6) (2010) 1634–1641.
[56] S. Barman, K. Srinivasan, Zinc supplementation alleviates hyperglycemia and as-
sociated metabolic abnormalities in streptozotocin-induced diabetic rats, Can. J.
Physiol. Pharmacol. (2016) ja.
[57] E. Ho, N. Quan, Y.-H. Tsai, W. Lai, T.M. Bray, Dietary zinc supplementation inhibits
NF κB activation and protects against chemically induced diabetes in CD1 mice,
Exp. Biol. Med. 226 (2) (2001) 103–111.
[58] Bestaoui A., ZINC ET PROFIL IMMUNO-INFLAMMATOIRE CHEZ LES ENFANTS ET
LES ADOLESCENTS DIABETIQUES DE TYPE 1, 2014.
[59] N.R. Pandey, G. Vardatsikos, M.Z. Mehdi, A.K. Srivastava, Cell-type-specific roles of
IGF-1R and EGFR in mediating Zn2+-induced ERK1/2 and PKB phosphorylation, J.
Biol. Inorg. Chem. 15 (3) (2010) 399–407.
[60] B. Li, W. Cui, Y. Tan, P. Luo, Q. Chen, C. Zhang, W. Qu, L. Miao, L. Cai, Zinc is
essential for the transcription function of Nrf2 in human renal tubule cells in vitro
and mouse kidney in vivo under the diabetic condition, J. Cell. Mol. Med. 18 (5)
(2014) 895–906.
[61] T. Liang, Q. Zhang, W. Sun, Y. Xin, Z. Zhang, Y. Tan, S. Zhou, C. Zhang, L. Cai, X. Lu,
Zinc treatment prevents type 1 diabetes-induced hepatic oxidative damage, en-
doplasmic reticulum stress, and cell death, and even prevents possible
M.M. Elseweidy et al.
steatohepatitis in the OVE26 mouse model: important role of metallothionein,
Toxicol. Lett. 233 (2) (2015) 114–124.
[62] W. Sun, X. Miao, S. Zhou, L. Zhang, P.N. Epstein, N. Mellen, Y. Zheng, Y. Fu,
Y. Wang, L. Cai, Zinc rescue of Akt2 gene deletion-linked murine cardiac dys-
function and pathological changes is metallothionein-dependent, J. Mol. Cell.
Cardiol. 74 (2014) 88–97.
[63] W. Sun, Y. Wang, X. Miao, Y. Wang, L. Zhang, Y. Xin, S. Zheng, P.N. Epstein, Y. Fu,
323
Biomedicine & Pharmacotherapy 95 (2017) 317–323
L. Cai, Renal improvement by zinc in diabetic mice is associated with glucose
metabolism signaling mediated by metallothionein and Akt, but not Akt2, Free
Radic. Biol. Med. 68 (2014) 22–34.
[64] M. Rafieian-kopaei, M. Gholami-Ahangaran, Comparison of disinfection activities of
nicotine with copper sulphate in water containing Limnatis nilotica. (OCAK-ŞUBAT)
(JANUARY-FEBRUARY), 2015: p. 9.