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Archives of Physiology and Biochemistry
ISSN: 1381-3455 (Print) 1744-4160 (Online) Journal homepage: http://www.tandfonline.com/loi/iarp20
Insulin and vanadium protect againstosteoarthritis development secondary to diabetesmellitus in rats
Abbas O. El Karib, Bahjat Al-Ani, Fahaid Al-Hashem, Mohammad Dallak,Ismaeel Bin-Jaliah, Basiouny El-Gamal, Salah O. Bashir, Refaat A. Eid &Mohamed A. Haidara
To cite this article: Abbas O. El Karib, Bahjat Al-Ani, Fahaid Al-Hashem, Mohammad Dallak,Ismaeel Bin-Jaliah, Basiouny El-Gamal, Salah O. Bashir, Refaat A. Eid & Mohamed A. Haidara(2016) Insulin and vanadium protect against osteoarthritis development secondary todiabetes mellitus in rats, Archives of Physiology and Biochemistry, 122:3, 148-154, DOI:10.3109/13813455.2016.1159698
To link to this article: http://dx.doi.org/10.3109/13813455.2016.1159698
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Arch Physiol Biochem, 2016; 122(3): 148–154! 2016 Informa UK Limited, trading as Taylor & Francis Group. DOI: 10.3109/13813455.2016.1159698
ORIGINAL ARTICLE
Insulin and vanadium protect against osteoarthritis developmentsecondary to diabetes mellitus in rats
Abbas O. El Karib1*, Bahjat Al-Ani1*, Fahaid Al-Hashem1, Mohammad Dallak1, Ismaeel Bin-Jaliah1,Basiouny El-Gamal2, Salah O. Bashir1, Refaat A. Eid3, and Mohamed A. Haidara1,4
1Department of Physiology, 2Department of Clinical Biochemistry, and 3Department of Pathology, College of Medicine, King Khalid University, Abha,
Saudi Arabia, and 4Department of Physiology, Kasr al-Aini Faculty of Medicine, Cairo University, Cairo, Egypt
Abstract
Objective: Diabetic complications such as cardiovascular disease and osteoarthritis (OA) areamong the common public health problems. The effect of insulin on OA secondary to diabeteshas not been investigated before in animal models. Therefore, we sought to determine whetherinsulin and the insulin-mimicking agent, vanadium can protect from developing OA in diabeticrats. Methods: Type 1 diabetes mellitus (T1DM) was induced in Sprague–Dawley rats and treatedwith insulin and/or vanadium. Tissues harvested from the articular cartilage of the knee jointwere examined by scanning electron microscopy, and blood samples were assayed foroxidative stress and inflammatory biomarkers. Results: Eight weeks following the induction ofdiabetes, a profound damage to the knee joint compared to the control non-diabetic groupwas observed. Treatment of diabetic rats with insulin and/or vanadium differentially protectedfrom diabetes-induced cartilage damage and deteriorated fibrils of collagen fibers. The relativebiological potencies were insulin + vanadium44insulin4vanadium. Furthermore, there wasabout 2- to 5-fold increase in TNF-a (from 31.02 ± 1.92 to 60.5 ± 1.18 pg/ml, p50.0001) and IL-6(from 64.67 ± 8.16 to 338.0 ± 38.9 pg/ml, p50.0001) cytokines and free radicals measured asTBARS (from 3.21 ± 0.37 to 11.48 ± 1.5mM, p50.0001) in the diabetic group, which wassignificantly reduced with insulin and or vanadium. Meanwhile, SOD decreased (from17.79 ± 8.9 to 8.250.29, p50.0001) and was increased with insulin and vanadium. The relativepotencies of the treating agents on inflammatory and oxidative stress biomarkers wereinsulin + vanadium44insulin4vanadium. Conclusion: The present study demonstrates that co-administration of insulin and vanadium to T1DM rats protect against diabetes-induced OApossibly by lowering biomarkers of inflammation and oxidative stress.
Keywords
Diabetes, T1DM, osteoarthritis, insulin,vanadium, rat model
History
Received 28 November 2015Revised 28 January 2016Accepted 24 February 2016Published online 23 March 2016
Introduction
An estimated 347 million people around the world suffer from
diabetes, which is one of the leading global health care
problems that claims the life of about 3.4 million every year
(Danaei et al., 2011). Type 1 diabetes mellitus (T1DM) is
believed to be caused by autoimmune antibodies targeting the
b cells-producing insulin in the pancreas that leads to the loss
of insulin production and hence the appearance of diabetes
symptoms in children, commonly between the ages of 7–19
years (Atkinson et al., 2014; Narendran et al., 2005; Pugliese,
2004).
Osteoarthritis (OA) is a degenerative joint disease that
involves degradation and destruction of the articular cartilage
structure of the joint leading to pain, swelling and reduced
joint movement; and there is increasing evidence to link
metabolic disturbances such as diabetes to OA (Pottie et al.,
2006, Van Manen et al., 2012). OA of the knee and/or hip is
regarded as one of the most prevalent conditions leading to
disability particularly in the elderly population (Grazio &
Balen, 2009). Knee OA is more important, not only for its
high prevalence rate compared with other types of OA, but
also for its presentation at earlier age groups particularly in
younger age groups of obese women (Bliddal & Christensen,
2009; Hayami, 2008).
Diabetic limited joint mobility syndrome is seen in 8–50%
of T1DM patients. The lack of the bone anabolic actions of
insulin and other pancreatic hormones could be the reason
why T1DM affects the skeleton more severely than type 2
diabetes mellitus (T2DM) (Hamann et al., 2012; Janghorbani
et al., 2007). In a study of patients with T2DM followed over
a course of 20 years it was concluded that longstanding
diabetes per se is detrimental for knee and hip joints, leading
to progressive destruction and joint failure (Schett et al.,
2013).
* These authors contributed equally to this work.
Correspondence: Professor Mohamed Haidara, Department ofPhysiology, College of Medicine, King Khalid University, Abha61421, Saudi Arabia. Tel: 00966540733723. E-mail: [email protected]
Proinflammatory biomarkers IL-6 and TNF-a are signifi-
cant predictors of knee OA (Kou & Wu, 2014; Livshits et al.,
2009) and OA cartilages from DM patients showed increased
responsiveness to IL-1b-induced inflammation via oxidative
stress and polyol pathway thus participate in the increased
inflammation in OA (Laiguillon et al., 2015).
Vanadium is a trace mineral that is widely distributed in
nature and has been reported to ameliorate DM in humans
when administered orally (Gaede et al., 2008). The glucose-
lowering effect of vanadium is attributed to the insulin-like
function of the compound and its ability to enhance insulin
activity and cellular bioavailability (Haidara et al., 2015;
Karmaker et al., 2007). Vanadium induces its effect through
insulin receptor-independent pathways since it has no effect in
activating the insulin receptor tyrosine kinase, a key enzyme
in insulin cell signalling (Shechter et al., 1995). However,
vanadium exerts its effects through inhibition of protein
tyrosine phosphatases, mainly PTEN and activation of
cytosolic tyrosine kinases that augment and activate down-
stream insulin receptor cell signalling (Srivastava & Mehdi,
2005). Studies have shown that vanadyl sulphate can lower
elevated blood glucose, cholesterol and triglycerides in a
variety of diabetic models including the streptozotocin (STZ)
diabetic rat, the Zucker fatty rat and the Zucker diabetic fatty
rat (Gruzewska et al., 2014).
Interestingly, neither the effect of insulin nor its mimicking
agent, vanadium on diabetes-induced OA in animal models
has been addressed before. Therefore, we were interested to
study the potential protective effect of insulin and or
vanadium against OA of the knee joint secondary to T1DM,
and to see if vanadium adjuvant treatment with insulin can
augment the proposed protection. To answer these questions,
we developed an animal model of T1DM-induced OA in
Sprague–Dawley rats and used the model to evaluate the
effect of insulin and vanadium on OA.
Methods
Animals
The experiments were performed on 36 male Sprague–
Dawley rats of 10 weeks old and weighting 200–250 g. The
rats were fed with standard laboratory diets, given water ad
libitum and maintained under laboratory conditions of
temperature (22 ± 3 �C), with 12-h light and 12-h dark
cycle. All experimental procedures involving the handling
and treatment of animals were approved by the Research
Ethical Committee of King Khalid University (Abha, KSA)
and were conducted in accordance with the National Institute
of Health’s Guide for the Care and Use of Laboratory
Animals.
Animal protocol
After 1-week adaptations, animals were classified and
randomly allocated into 6 groups (n¼ 6) as follows: control
group (Control): non-diabetic, non-treated rats that were
injected intraperitoneally (i.p.) once with citrate buffer (0.1 M,
pH 4.5); vanadium-treated group (Van.): non-diabetic, van-
adium-treated rats that were i.p. injected with citrate buffer as
control group, and received vanadyl sulphate of 0.64 mmol/kg
body weight, freshly dissolved in 1 ml of distilled water daily
through a nasopharyngeal tube (Haidara et al., 2014);
Diabetic type 1 group (D1): DM was induced in rats by a
single i.p. injection of STZ in a dose of 65 mg/kg. Rats
showed glucose level above 300 mg/dl were considered to be
diabetics and included in the experiments (El Karib, 2014;
Haidara et al., 2004); T1DM and insulin group (D1 + Ins.):
rats were made type 1 diabetic as in T1DM group and
received Mixtard insulin subcutaneously in a dose of 0.75 IU/
100 g weight in 0.75 ml volume once daily (Haidara et al.,
2015), after 48 h of diabetic induction; T1DM and vanadium
group (D1 + Van.): T1DM rats received the same dose of
vanadium as in vanadium group after 48 h of induction of
T1DM; T1DM and vanadium and insulin group
(D1 + Van. + Ins.): T1DM rats received both vanadium and
insulin with the same doses as in previous groups.
At the end of the 8th weeks of the experiment, 5 ml retro-
orbital blood samples was obtained under anaesthesia using
40 mg kg�1 sodium thiopentane, i.p., after fasting. Blood was
collected into plain tubes, then allowed to clot for 20 min then
centrifuged at 14 000 rpm for 10 min for serum separation.
Then sera were stored at �80 �C, for subsequent measure-
ments of biochemical parameters. After withdrawal of the
blood samples, the knee joints were opened, dissected, and
fixed in glutaraldehyde and kept for electron microscopy
examinations.
Measurement of IL-6, TNF-a cytokines
Rat interleukin-6 ELISA kit, IL-6 for serum, plasma and
tissue culture supplements (Ray Biotech Inc., Mfr. No. ELR-
IL6-001) and rat tumour necrosis factor alpha (TNF-a ELISA
kit BIOTANG Inc., Cat. No. R6365 were used as recom-
mended by the manufacturer.
Measurements of SOD and TBARS
Superoxide dismutase (SOD) assay kit, rat, Cayman
Chemical, Cat. No. 706002 and thiobarbituric acid reactive
substances (TBARS) assay kit, Cayman Chemical, Item
Number 10009055 were used as recommended by the
manufacturer.
Scanning electron microscopy
Different specimens were taken from articular cartilage of
all treated rats and subjected for ultrastructural examin-
ations with scanning electron microscope. Ultra-structural
micrographs are representative of the changes seen in
different treated groups. The specimens were fixed with
2.5% (wt/vol) sodium cacodylate-buffered glutaraldehyde,
pH 7.2 at 4 �C for 2 h. Specimens were also post fixed in
1% sodium cacodylate-buffered osmium tetroxide, pH 7.2
for 1 h. After washing and dehydration in ascending series
of ethanol, critical-point drying was performed using the
EMITECH-K850 critical-point drying unit. The specimens
were mounted on aluminium stubs with double-sided tape
and silver glue and then sputter coated with gold by BOC
EDWARDS SCANCOAT. The specimens were observed
using a Jeol field emission scanning electron microscope
JSM-6390LV.
DOI: 10.3109/13813455.2016.1159698 Insulin and vanadium protect against OA 149
Statistical analysis
Data are presented as mean ± SD. Comparison of data was
analysed using Graph pad Prism software, version 5.
Comparison between the groups was performed by one-way
analysis of variance, followed by Tukey’s post-hoc test.
Probability (p) values of 50.05 were considered to be
significant.
Results
Changes in blood glucose level
Figure 1 shows a significant (p50.0001) increase in blood
glucose levels in diabetic group (371 ± 27 mg/dl) in compari-
son to the control group (92 ± 5 mg/dl). Compared to diabetic
group, administration of vanadium and or insulin caused a
significant (p50.0001) decrease in glucose levels. In add-
ition, insulin alone or with vanadium, but not vanadium alone,
were able to lower blood glucose to levels comparable to the
control group.
Osteoarthritis development in rats by induction ofdiabetes
Eight-week post-induction of diabetes, tissue preparations for
histological examination under scanning electron microscopy
(Figures 2 and 3) from the articular cartilage of the knee joint
of the sacrificed T1DM rats (Figures 2C and 3C) revealed OA
development compared to the non-diabetic healthy control
rats (Figures 2A and 3A). The pathological changes revealed
a massive destruction of the articular cartilage caused by
T1DM such as atrophic chondrocytes inside distorted lacunae
that are surrounded by damaged territorial matrix
(Figure 2C), compared to normal structures in control tissues
(Figure 2A). Furthermore, scanning electron micrographs of
T1DM rats’ collagen fibers showed extensive damage and
deterioration in fibrils forming sheets (Figure 3C), whereas
normal fibrils forming sheets of collagen fibers were depicted
in control rats (Figure 3A).
Insulin and vanadium protect the knee joint againstT1DM-induced osteoarthritis
A substantial protection was obtained when insulin was given
to diabetic rats (Figures 2D and 3D). 48 h after induction of
diabetes, treating diabetic group with insulin for 8 weeks
protected the OA ultrstructure of the articular cartilage to
become much less damaged comparable to the control group.
Scanning electron micrographs of insulin-treated tissues
showed a normal territorial matrix and restoration of the
leaving lacunae most of its normal size and regular shape,
with a clear improvement in the healing of chondrocytes
(Figure 2D). In addition, collagen fibers showed intact fibril
sheets, but still with some focal damage (Figure 3D).
Administration of diabetic group with vanadium for 8
weeks differentially protected the ultrastructure of the articu-
lar cartilage (Figures 2E and 3E). Electron microscopy results
revealed a better protection of leaving lacunae and
territorial matrix than chondrocytes and collage fibers
(Figures 2E and 3E). However, focal damage of leaving
lacunae and atrophic chondrocytes are still shown. Strikingly,
co-administration of insulin and vanadium protects the
articular cartilage of the diabetic rats where it showed
normal chondrocytes, leaving lacunae, territorial matrix and
collagen fibres (Figures 2F and 3F). The above treatments
were compared to control groups, non-diabetic ‘‘vehicle-
treated’’ (Figures 2A and 3A) and non-diabetic ‘‘vanadium-
treated’’ rats (Figures 2B and 3B) that showed normal
structures of the articular cartilage.
Insulin and vanadium inhibit T1DM-induced cytokinesand oxidative stress
Proinflammatory cytokines are known to be involved in the
pathology of diabetes (Senn et al., 2002; Shoelson et al.,
2006; Southern et al., 1990) and OA (Doss et al., 2007,
Stannus et al., 2010). To investigate whether vanadium and/or
insulin can suppress the release of the proinflammatory
cytokines in T1DM, we measured the blood level of TNF-aand IL-6, 8 weeks after T1DM induction. T1DM caused a 2-
fold increase in TNF-a and 4-fold increase in IL-6 that were
significantly inhibited by insulin or vanadium or vanadium
plus insulin (Figure 4A and B).
Administration of insulin and vanadium significantly
decreased IL-6 in comparison either to diabetic rats admin-
ister either insulin (p50.0001) or vanadium (p50.001)
alone. Using post-hoc testing, least significant difference
(LSD) for IL-6, it was found that there was a significance
difference between diabetic group that received insulin and
vanadium in comparison to either diabetes and insulin
(p50.0001) or diabetes and vanadium (p50.0001) alone.
Insulin and vanadium administration to diabetic group
significantly decreased TNF-a in comparison either to
diabetic rats administer either insulin (p50.0001) or van-
adium (p50.0001) alone. Using post-hoc testing (LSD) for
Figure 1. Changes in blood glucose level. Control, control vehicleinjected rats; Van., vanadium-treated non-diabetic rats; D1, streptozoto-cin-injected rats (type 1 diabetic group); D1 + Ins., diabetic group treatedwith insulin; D1 + Van., diabetic group treated with vanadium;D1 + Van. + Ins., diabetic group treated with vanadium and insulin. Allresults are the mean (±SD) of three experiments. Significance indicated:p50.05, P1: significant in comparison to control group, P2: significantin comparison to vanadium group, P3: significant in comparison todiabetic group.
150 A. O. El Karib et al. Arch Physiol Biochem, 2016; 122(3): 148–154
TNF-a, it was found that there was a significance difference
between diabetic group that received insulin and vanadium in
comparison to either diabetes and insulin (p50.0001) or
diabetes and vanadium (p50.0001) alone.
To determine whether insulin and/or vanadium can
modulate the oxidant and anti-oxidant biomarkers known to
be affected by diabetes (Molinar-Toribio et al., 2014) and OA
(Gutteridge, 1995), we measured the free radicals as TBARS,
and SOD as biomarkers of oxidative stress by ELISA. Our
results showed that diabetic induction augmented TBARS and
suppressed SOD (Figure 5A and B). Insulin significantly
augmented SOD and inhibited TBARS, whereas vanadium on
its own was a weak stimulant to SOD and exerted no
significant inhibitory effect on TBARS levels in T1DM group.
Furthermore, co-administration of insulin with vanadium
completely restored the levels of SOD and TBARS to the
control levels (Figure 5A and B).
Administration of insulin and vanadium significantly
decreased TBARS in comparison either to diabetic rats
administer either insulin (p50.0001) or vanadium
(p50.0001) alone. Using post-hoc testing (LSD) for
TBARS, it was found that there was a significance difference
between diabetic group that received insulin and vanadium in
comparison to either diabetes and insulin (p50.0001) or
diabetes and vanadium (p50.0001) alone.
Insulin and vanadium administration to diabetic group
significantly increased SOD in comparison either to diabetic
rats administer either insulin (p50.0001) or vanadium
(p50.0001) alone. Using post-hoc testing (LSD) for SOD,
it was found that there was a significance difference between
diabetic group that received insulin and vanadium in
comparison to either diabetes and insulin (p50.0001) or
diabetes and vanadium (p50.0001) alone.
Discussion
The principal finding of our study was that insulin in
combination with the insulinomimetic agent, vanadium
inhibited OA induced by T1DM and suppressed the bio-
markers of inflammation and oxidative stress known to be
induced in diabetes and OA. Furthermore, insulin on its own
was more potent than vanadium to protect against OA. To the
best of our knowledge, these studies are the first to report a
clear protective role for insulin and vanadium in OA induced
Figure 2. Insulin and vanadium protected theknee joint against OA in diabetic rats.Scanning electron micrographs (2000�) ofrats’ articular cartilage from the knee joint 8weeks after the induction of diabetes bystreptozotocin. A. Control, control vehicleinjected rats. B. Van., vanadium-treated non-diabetic rats. C. D1, streptozotocin-injectedrats (type 1 diabetic group). D. D1 + Ins.,diabetic group treated with insulin. E.D1 + Van., diabetic group treated with van-adium. F. D1 + Van. + Ins., diabetic grouptreated with vanadium and insulin.Abbreviations: C, chondrocyte; C1, focaldamage of chondrocyte; L, leaving lacunae;N, nuclei; TM, territorial matrix. Note thatarrows point to atrophic chondrocyte shownin (D) and focal damage of leaving lacunaein (E).
DOI: 10.3109/13813455.2016.1159698 Insulin and vanadium protect against OA 151
Figure 3. Insulin and vanadium protectedcollagen fibres in the knee joint against OA indiabetic rats. Scanning electron micrographs(2000�) of rats’ collagen fibres in articularcartilage from the knee joint 8 weeks after theinduction of diabetes by streptozotocin. A.Control, control vehicle injected rats. B. Van.,vanadium-treated non-diabetic rats. C. D1,streptozotocin-injected rats (type 1 diabeticgroup). D. D1 + Ins., diabetic group treatedwith insulin E. D1 + Van., diabetic grouptreated with vanadium. F. D1 + Van. + Ins.,diabetic group treated with vanadium andinsulin. Abbreviations: F, normal or intactcollagen fibrils sheets; F1, damaged collagenfibrils sheets. Note that arrows point to focaldamage of collagen fibril sheets.
Figure 4. Insulin and vanadium inhibit TNF-a and IL-6 in T1DM-treated rats. A, TNF-a and (B) IL-6 levels in blood from the groups (n¼ 6) weremeasured after 8-week treatment by ELISA. Control, control vehicle injected rats; Van., vanadium-treated non-diabetic rats; D1, streptozotocin-injectedrats (type 1 diabetic group); D1 + Ins., diabetic group treated with insulin; D1 + Van., diabetic group treated with vanadium; D1 + Van. + Ins., diabeticgroup treated with vanadium and insulin. All results are the mean (±SD) of three experiments. Significance indicated: p50.05, P1: significant incomparison to control group, P2: significant in comparison to vanadium group, P3: significant in comparison to diabetic group.
152 A. O. El Karib et al. Arch Physiol Biochem, 2016; 122(3): 148–154
by diabetes. These conclusions were supported by the data
indicating that STZ induced T1DM in rats caused a substan-
tial destruction to the articular cartilage of the knee joint that
was protected by insulin and vanadium.
Studies on experimental animals have demonstrated that
administration of STZ, depending on its dose, extensively
reduces beta cells mass and destroys pancreatic islet (Cam
et al., 1997; Yanardag et al., 2005). Our results showed that
vanadium and insulin administration causes decrease glu-
cose level in diabetic group to control level which is in
accordance with previous work that showed that regardless
of the severity of diabetes, vanadium therapy needs a
minimum level of plasma insulin, secreted endogenously, or
received exogenously to assist vanadium to induce normo-
glycemia (Pirmoradi et al., 2014; Yanardag et al., 2005). In
addition, the suppression of the growth body weight seen in
the diabetic group was linked with the impaired carbohy-
drate metabolism as a source of energy (Pires et al., 2014,
Wycherley et al., 2014). However, the suppressive action of
vanadium on the appetite, via anorexigenic stimulation of
the central nervous system and food restriction, was
possibly the cause of reduced growth body weight in the
vanadium-treated group (Marty et al., 2007). This is in
agreement with our findings that showed no significant
changes in the body weight of diabetic or vanadium-treated
groups (data not shown).
We confirmed by electron microscopy examination the
development of OA in the articular cartilage of the knee joint,
which is known to be targeted by diabetes (El Karib, 2014;
Musumeci et al., 2014; Niethard, 1986; Pottie et al., 2006).
Tissue examination revealed a profound destruction of the
cartilage structures such as chondrocytes, territorial matrix
and collagen fibres.
Prolonged hyperglycemia and accumulation of glucose
derived advanced glycation end products (AGEs) were
proposed to contribute to OA development and insulin is
reported to have anabolic effects on bone (Yan & Li, 2013). In
addition, insulin was found to increase proteoglycan synthesis
in primary chondrocytes tissue culture and increased proteo-
glycan and matrix synthesis in articular cartilage explants
obtained from healthy pigs and diabetic human and mice (Cai
et al., 2002). We therefore tested to see if insulin and its
mimicking agent, vanadium is able to prevent articular
cartilage injury in animals following diabetes. Indeed, our
data showed that giving insulin or vanadium partially
protected the knee joint injury. However, when both insulin
and vanadium were given together, they protected the
cartilage and the histological pictures were similar to the
non-diabetic controls. Furthermore, It is important to note that
the relative potencies for these agents in our T1DM-induced
OA animal model were determined to be insulin + van-
adium44insulin4vanadium. Our data also point to the
importance of co-administering insulin and vanadium in
preventing the increase of inflammatory and oxidative
biomarkers levels above the control in diabetic-treated rats
(Figures 4 and 5). It was also interesting to report that the
relative potencies of vanadium were observed here to be well
below that of insulin, which is in agreement with potent
insulin over vanadium that we observed in the healing of the
damaged cartilage.
Our protective approach, by co-administrating insulin and
vanadium, that prevents the diabetic rats to develop OA
concomitant with a complete inhibition of TNF-a, IL-6 and
free radicals points to these cytokines and TBARS oxidative
stress as very important proinflammatory biomarkers to
monitor OA secondary to diabetes, including the healing
progress of the disease. Indeed, our findings are in agreement
with the previously published work on the role of TNF-a and
IL-6 in the pathogenesis of inflammation induced bone loss,
osteoclastic bone resorption and cartilage destruction (Fuller
et al., 2002; Kapoor et al., 2011; Korczowska & Lacki, 2005)
and the reports that proposed IL-6 and TNF-a as significant
predictors of knee OA (Kou & Wu 2014; Livshits et al.,
2009).
Taken together, our data support the conclusion that insulin
protects against development of OA induced by T1DM and
co-administration of vanadium with insulin is more effective
to protect against the diabetic OA complications in STZ-
induced OA in this rat model by lowering biomarkers of
inflammation and levels of oxidative stress.
Figure 5. Different effects of insulin and vanadium on inhibition of TBARS and activation of SOD in T1DM-treated rats. (A) TBARS and (B) SODlevels in blood from the 6 rat groups were measured after 8-week treatment by TBARS assay and ELISA. Control, control vehicle injected rats; Van.,vanadium-treated non-diabetic rats; D1, streptozotocin-injected rats (type 1 diabetic group); D1 + Ins., diabetic group treated with insulin; D1 + Van.,diabetic group treated with vanadium; D1 + Van. + Ins., diabetic group treated with vanadium and insulin. All results are the mean (±SD) of threeexperiments. Significance indicated: p50.05, P1: significant in comparison to control group, P2: significant in comparison to vanadium group, P3:significant in comparison to diabetic group.
DOI: 10.3109/13813455.2016.1159698 Insulin and vanadium protect against OA 153
Declaration of interest
All authors declare that they have no conflict of interest.
This work was supported by grants of King Khalid
University. KKU-Project No 2 (215).
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