DM Induction Research Articles

Beneficial Effect of Sirolimus-Pretreated Mesenchymal Stem Cell Implantation on Diabetic Retinopathy in Rats.

Diabetic retinopathy (DR) is a vision-threatening complication that affects virtually all diabetic patients. Various treatments have been attempted, but they have many side effects and limitations. Alternatively, stem cell therapy is being actively researched, but it faces challenges due to a low cell survival rate. In this study, stem cells were pretreated with sirolimus, which is known to promote cell differentiation and enhance the survival rate. Additionally, the subconjunctival route was employed to reduce complications following intravitreal injections. Diabetes mellitus was induced by intraperitoneal injection of 55 mg/kg of streptozotocin (STZ), and DR was confirmed at 10 weeks after DM induction through electroretinogram (ERG). The rats were divided into four groups: intact control group (INT), diabetic retinopathy group (DR), DR group with subconjunctival MSC injection (DR-MSC), and DR group with subconjunctival sirolimus-pretreated MSC injection (DR-MSC-S). The effects of transplantation were evaluated using ERG and histological examinations. The ERG results showed that the DR-MSC-S group did not significantly differ from the INT in b-wave amplitude and exhibited significantly higher values than the DR-MSC and DR groups (p < 0.01). The flicker amplitude results showed that the DR-MSC and DR-MSC-S groups had significantly higher values than the DR group (p < 0.01). Histological examination revealed that the retinal layers were thinner in the DR-induced groups compared to the INT group, with the DR-MSC-S group showing the thickest retinal layers among them. Subconjunctival injection of sirolimus-pretreated MSCs can enhance retinal function and mitigate histological changes in the STZ-induced DR rat model.

Mechanistic Insights of Meglitinides Drug Repurposing and Delivery to Brain for Metabolic Syndrome Linked Neurodegenerative Disease

AbstractBackgroundMetabolic syndrome (MS) includes clusters of metabolic disorders and type 2 DM (T2DM) is most common MS which cause insulin resistance (IR). The IR mediated dysregulation of signaling cascades in brain increase the production and secretion of Aβ, p‐tau which prompt to Alzheimer’s (AD). Therefore, repaglinide (REP) a meglitinides drug is used, which act by targeting ATP binding cassette and evidence indicated that REP increase neuronal survival via upregulating ATF‐6 gene and blocking the DREAM. Thus, there is huge possibility that REP regulate and modulate the expression of pro‐apoptotic protein, anti‐apoptotic protein, calcium homeostasis and may eventually reduce the neuronal cell death.MethodsIn, in vitro studies, primarily cytotoxicity assay was performed on SHSY‐5Y cells and to mimic the neurodegenerative condition, the cells were treated with streptozotocin (STZ, 2.5mM). Further, to check the efficacy of REP in AD, in vivo studies were performed using HFD fed STZ induced murine model. The HFD was fed for 16 weeks followed by administration of STZ (30mg/kg; i.p) in wistar rats for induction of DM linked AD.ResultsIn vitro and in vivo studies were conducted to understand the neuroprotective potential of the REP. It was observed that the REP shows cellular viability of ∼80‐85% and no morphological changes in the cells were identified. Additionally, the results have shown that REP increase the cell viability of STZ treated SHSY‐5Y cells by ∼1.2‐fold and significantly (p&lt;0.01) decreased the oxidative stress in H2O2 treated cells. After treatment of REP (4 mg/kg,p.o) for 4 weeks in DM linked AD wistar rats, a significant (p&lt;0.01) decrement in Aβ, tau proteins, TNF‐α, IL‐6, MDA, NO and increment in BDNF, SOD, GSH level was observed compared to disease control rats. Further, in behavioral studies a significant improvement (p&lt;0.01) in the retention memory and spatial memory was observed after the REP treatment. Additionally, a significant (p&lt;0.05) increase in Bcl‐2, ATF‐6 expression, and decrement in Bax, Caspase‐3 expression indicates reduction in neuronal cell death then the disease rats.ConclusionIn summary, it concluded that REP act as a neuroprotective agent which might be used to treat MS linked AD.

The Effects of Metformin Treatment on Diabetic Albino Rats' Pancreas, Liver, and Kidney Histology.

Metformin, an oral hypoglycemic drug, has traditionally been considered the standard therapy for hyperglycemia. Metformin's several modes of action include inhibition of hepatic gluconeogenesis, anti-glucagon activity, and insulin-sensitizing effect. This study aims to assess the effectiveness of Metformin on the liver, pancreatic, and kidney tissues of alloxan-induced diabetic albino rats. Twenty mature albino white male rats were allocated at random into two groups. Intraperitoneal injections of alloxan monohydrate were utilised to induce diabetic Mellitus type II in the first ten rats. The second group of rats were injected intraperitoneally with normal saline. Both groups were then separated into four subgroups: Group 1 consisted of non-diabetic rats that were only administered distilled water (control), Group 2 consisted of non-diabetic rats that were administered metformin at a dose of 1000 mg/kg/day, and Group 3 consisted of diabetic control animals that were administered alloxan intravenously and distilled water orally, but were not given any medications. After seven days of DM induction, diabetic rats were administered Metformin at a dose of 1000 mg/kg/day orally. After one month of therapy, the animals were slaughtered and their organs were harvested. Compared to the control group, the histological results of pancreatic tissue were normal in the treatment groups. In contrast, liver and kidney sections from non-diabetic control, non-diabetic, and diabetic animals given 1000 mg/kg/day of Metformin had normal histology. Still, both tissues of untreated diabetic control mice exhibited lymphocyte infiltration. Metformin has been found to have significant blood glucose lowering properties and the capacity to protect several organs from the negative consequences of diabetes.

Abstract 111: Ischemic Post Conditioning After Asphyxial Arrest In A Diabetic Rat Model

Diabetes (DM) is a pervasive disease associated with increased risk of cardiac arrest (CA) and worse survival. Diabetics display abrogation of post arrest therapies. Ischemic post-conditioning (IPoC) has been shown to improve outcomes in young healthy swine, but never in a DM model. We hypothesized that IPoC would be beneficial for survival of non-DM rats after asphyxial CA and that this benefit would be abrogated in DM. Male Wistar rats were utilized, 12 of 23 underwent induction of DM with injection of streptozotocin (30mg/kg) IP and feeding of western diet for 12 weeks. Rats were intubated, mechanically ventilated, and anesthetized with isoflurane. A non-invasive flow probe was placed on a carotid artery. An internal jugular vein and femoral artery and vein were cannulated. Electrocardiogram, echocardiogram, and arterial blood gas were analyzed. Asphyxial CA, induced by rocuronium and cessation of ventilation, proceeded for 7 min. CPR was initiated utilizing an automated chest compressor at a rate of 200/min. Animals were randomly assigned to standard CPR (S-CPR) or IPoC. IPoC was initiated 40 seconds into CPR with a 20 sec pause followed by 20 sec of compressions, 1 cycle. This cycling was performed 4 total times after which S-CPR was continued. Defibrillation and epinephrine were given, as necessary. Data were collected every 40 sec during CPR and at 15 min, 1 and 2 hr after return of spontaneous circulation (ROSC). Data were analyzed using ANOVA with pairwise comparisons. Significance set at p &lt;.05. Non-DM rats displayed high rates of ROSC in both S-CPR, and IPoC groups, 100% (5 of 5) and 83% (5 of 6) respectively. DM S-CPR rats had a reduced ROSC rate at 14% (1 of 7) which improved to 80% (4 of 5) with IPoC. There were no significant differences between DM S-CPR and IPoC groups during CPR. Only DM rats required defibrillations. DM rats had a significantly longer time to ROSC, higher epinephrine requirement, lower nadir pH, and lower carotid blood flow at end experiment. There was no difference in any marker between non-DM S-CPR and IPoC groups after ROSC. DM was associated with increased injury from CA. IPoC was not harmful to non-DMs but improved ROSC rate and thus survival in DM rats. This is a novel demonstration of IPoC utility in a historically conditioning resistant group.

Altered Expression of EMT-Related Factors Snail, Wnt4, and Notch2 in the Short-Term Streptozotocin-Induced Diabetic Rat Kidneys.

Background: The aim of this study was to determine the expression of epithelial to mesenchymal transition (EMT)-related transcription factors Snail, Wnt4, and Notch2 with key roles in renal fibrosis, in different renal areas of diabetic rats: glomeruli (G), proximal and distal convoluted tubules (PCT; DCT). Methods: Male Sprague Dawley rats were instilled with 55 mg/kg streptozotocin (diabetes mellitus type I model, DM group) or citrate buffer (control group). Kidney samples were collected 2 weeks and 2 months after DM induction and processed for immunohistochemistry. Results: Diabetic animals showed higher Wnt4 kidney expression both 2 weeks and 2 months post-DM induction, while Snail expression significantly increased only 2 weeks after DM initiation (p < 0.0001). We determined significantly higher expression of examined EMT-related genes in different kidney regions in diabetic animals compared with controls. The most substantial differences were observed in tubular epithelial cells in the period of 2 weeks after induction, with higher Snail and Wnt4 expression in PCT and increased Snail and Notch2 expression in DCT of diabetic animals (p < 0.0001; p < 0.001). Conclusion: The obtained results point to the EMT-related factors Snail, Wnt4, and Notch2 as a potential contributor to diabetic nephropathy development and progression. Changes in their expression, especially in PCT and DCT, could serve as diagnostic biomarkers for the early stages of DM and might be a promising novel therapeutic target in this condition.

The potential role of LCZ696 in ameliorating diabetic cardiomyopathy by inhibiting the myocardial inflammation, ER stress, and apoptosis through the RAGE/NF‐κB and PERK/CHOP signaling Pathways

The development of diabetic cardiomyopathy (DCM) is multifactorial that leads to myocardium structural and functional disruption resulting excessive activity of the renin‐angiotensin system (RAS) and finally heart failure. LCZ696 newly established as chronic heart failure drug which combines a neprilysin inhibitor (sacubitril) and an angiotensin II receptor blocker (valsartan). Present study is designed to find the effect of LCZ696 on DCM in rats. Diabetes was induced by feeding rats with a high‐fat diet for 6 weeks following a single injection of STZ (30mg/kg). Diabetic rats were randomly divided in three group (n=8), LCZ696 and valsartan treatment was started two weeks after the DM induction and continued for 8 consecutive weeks. Blood pressure of individual rats was recorded using tail‐cuff instruments. At end of the treatment echocardiography was done. In serum glucose, insulin, creatine kinase‐MB and lactate dehydrogenase (LDH). In cardiac tissues, pro‐inflammatory cytokines and advanced glycation end‐products (AGEs) formation estimated. Western blot and RT‐PCR were used to determine the apoptotic genes, endoplasmic reticulum stress (ER) parameters. LCZ696 and valsartan ameliorated the DCM progression by inhibiting of AGEs, its receptor (RAGE) at mRNA, pro‐apoptotic gene markers (BAX and caspase‐3) associated with induction of the antiapoptotic gene (Bcl‐2) at mRNA and protein expressions, NF‐κB at the mRNA and protein levels associated with restoring the elevation of pro‐inflammatory cytokines such as TNF‐α, IL‐6, and IL‐1β at the activity level. Furthermore, LCZ696 and valsartan helps in restoring the induction of ER stress parameters (GRP78, PERK, eIF2a, ATF4, and CHOP) at the mRNA and protein levels. LCZ696 and valsartan improved diastolic dysfunction and attenuated histological abnormalities. LCZ696 and valsartan attenuated DCM by inhibiting the myocardial inflammation, ER stress, and apoptosis through the RAGE/NF‐κB and PERK/CHOP signaling cascades. Collectively, present results revealed LCZ696 has more strong protective effect against DCM than valsartan.

Impaired pulmonary vasomotor control in exercising swine with multiple comorbidities

Pulmonary hypertension is common in heart failure with preserved ejection fraction (HFpEF). Here, we tested the hypothesis that comorbidities [diabetes mellitus (DM, streptozotocin), hypercholesterolemia (HC, high-fat diet) and chronic kidney disease (CKD, renal microembolization)] directly impair pulmonary vasomotor control in a DM + HC + CKD swine model. 6 months after induction of DM + HC + CKD, pulmonary arterial pressure was similar in chronically instrumented female DM + HC + CKD (n = 19) and Healthy swine (n = 18). However, cardiac output was lower both at rest and during exercise, implying an elevated pulmonary vascular resistance (PVR) in DM + HC + CKD swine (153 ± 10 vs. 122 ± 9 mmHg∙L−1∙min∙kg). Phosphodiesterase 5 inhibition and endothelin receptor antagonism decreased PVR in DM + HC + CKD (− 12 ± 12 and − 22 ± 7 mmHg∙L−1∙min∙kg) but not in Healthy swine (− 1 ± 12 and 2 ± 14 mmHg∙L−1∙min∙kg), indicating increased vasoconstrictor influences of phosphodiesterase 5 and endothelin. Inhibition of nitric oxide synthase produced pulmonary vasoconstriction that was similar in Healthy and DM + HC + CKD swine, but unmasked a pulmonary vasodilator effect of endothelin receptor antagonism in Healthy (− 56 ± 26 mmHg∙L−1∙min∙kg), whereas it failed to significantly decrease PVR in DM + HC + CKD, indicating loss of nitric oxide mediated inhibition of endothelin in DM + HC + CKD. Scavenging of reactive oxygen species (ROS) had no effect on PVR in either Healthy or DM + HC + CKD swine. Cardiovascular magnetic resonance imaging, under anesthesia, showed no right ventricular changes. Finally, despite an increased contribution of endogenous nitric oxide to vasomotor tone regulation in the systemic vasculature, systemic vascular resistance at rest was higher in DM + HC + CKD compared to Healthy swine (824 ± 41 vs. 698 ± 35 mmHg∙L−1∙min∙kg). ROS scavenging induced systemic vasodilation in DM + HC + CKD, but not Healthy swine. In conclusion, common comorbidities directly alter pulmonary vascular control, by enhanced PDE5 and endothelin-mediated vasoconstrictor influences, well before overt left ventricular backward failure or pulmonary hypertension develop.

Effects of renal denervation on cardiovascular, metabolic and renal functions in streptozotocin-induced diabetic rats

Diabetes promotes renal sympathetic hyperactivity, autonomic imbalance, and cardiovascular and renal dysfunction. Bilateral renal denervation (BRD) has emerged as a treatment for diabetes; however, the mechanisms that underlie the beneficial effects of BRD are unknown. AimsThe present study evaluated the effects of BRD on autonomic, cardiovascular, metabolic, and renal function in streptozotocin-diabetic rats. Main methodsWistar rats were separated into three experimental groups: control (CTR), diabetic (DM), and diabetic that underwent BRD (DM BRD). BRD was performed two weeks after STZ-diabetes induction, the experiments were performed four weeks after DM induction. This study evaluated sympathetic vasomotor nerve activity in different territories (renal, lumbar and splanchnic), arterial baroreceptor reflex, metabolic and renal function. Key findingsBRD significantly reduced glycemia, glycosuria, albuminuria, and SGLT2 gene expression in the kidney in DM rats. Renal sympathetic nerve activity (rSNA) was significantly increased and splanchnic sympathetic nerve activity (sSNA) was significantly decreased in DM rats, without changes in lumbar sympathetic nerve activity (lSNA). BRD was able to normalize sSNA and significantly increase lSNA in DM rats compared to control rats. Additionally, cardiac baroreceptor sensitivity was impaired in DM rats, and BRD significantly improved baroreflex sensitivity. SignificanceOur data suggest that renal nerves play an important role in autonomic, cardiovascular, and renal dysfunction in STZ-DM rats. Thus, sympathetic renal hyperactivity should be considered a possible therapeutic target in diabetic patients.

Role of Brain Derived Neurotrophic Factor and Swimming Exercise in Cardiac Autonomic Neuropathy in Type 2 Diabetes Rat Model

Cardiac Autonomic Neuropathy (CAN) is a major chronic complications of DMII. Clinical investigations have revealed that uncontrolled DM II is frequently associated with lower levels of BDNF. This study assessed whether diminished BDNF level is the reason of CAN caused by DM II and if swimming aerobic exercise can increase BDNF level to ameliorate that CAN . Sixty rats were divided into six groups. Group I; Normal (N), Group 2; BDNF blocker, Group 3; DM II, Group 4; DM II + BDNF, Group 5; DM II + Swimming exercise and Group 6; DM II + Swimming exercise + BDNF blocker. Induction of DM II was done by intraperitoneal injection of 180 mg/kg of Nicotinamide 30 minutes before injection of 50 mg/kg Streptstozin. Swimming exercise began with a 1-week adaptation with gradual increase of swimming duration until reaching 1h swimming/day at the end of the 1st week. Then, a 7-weeks training program with 5 days/week swimming was followed. The duration of the experiment was 8 weeks during which the following parameters were measured: body weight, random blood glucose “RBG” level, plasma insulin level, heart rate “HR”, systolic blood pressure “SBP”, heart rate variability “HRV”, baroreflex sensitivity “BRS”, vascular sympathetic tone “VST” and plasma BDNF. Results revealed that BDNF significantly improved RBG level, HRV, VMT and BRS. Swimming exercise significantly increases plasma BDNF level and also improved CAN. Conclusively, BDNF can be used as a treatment regimen for CAN. Also swimming exercise has great therapeutic effect on CAN maybe by producing BDNF.

FUNCTIONAL ANALYSIS OF GAIT WITH INSOLE USE IN DIABETIC FEMALE RATS: A CROSS-SECTIONAL STUDY

ABSTRACT Introduction: Diabetes mellitus is a chronic disease that is characterized by causing damage to the peripheral nervous system, generating sensory and motor changes. Objective: This study aims at analyzing the impact of the use of different orthotic insoles on the gait of diabetic female rats. Methods: Twenty-six female Wistar rats were randomly divided into the Control and Diabetic groups. The mechanical sensitivity test was performed manually on the surface of the animals' hind paws using the von Frey test. The functional evaluation was carried out on an adapted platform where the animals were stimulated to walk in order to capture images of the ventral region for measurements of the right and left hind paws. After the images were collected they were processed using Kinovea software version 0.8.27 to assess: stride distance, time, speed and acceleration. Results: There was a reduction in the weight of the animals in the Diabetic Group (p = 0.0018), associated with hyperglycemia (p = &lt;0.0001), and a decrease in mechanical sensitivity as compared to the Control Group (p = 0.0372). Gait analysis showed a reduction in stride speed (p = 0.0482) and acceleration (p = 0.0149), with the silicone orthosis in the Diabetic Group. Conclusions: The silicone orthosis demonstrated a reduction in stride speed and acceleration, without compromising the other variables in the diabetic rats. The other insoles showed no functional difference between groups. Even though the animals showed a change in sensitivity at the end of 28 days of DM induction, this time does not appear to have been able to develop extensive changes in the rats' gait function. Level of evidence II; Therapeutic studies - Investigating the Results of Treatment.

SDR 기반 자기 유도 OFDM 통신 시스템 구현

자기 유도 통신은 환경에 따른 영향에 민감하지 않아서 음파, 광파, 전자파 등이 잘 전달되지 않는 수중, 지하, 파이프라인 등의 복잡한 환경에서의 통신을 위해 최근 들어 큰 관심을 받고 있다. 하지만 자기 유도 통신 시스템은 코일 안테나 회로로 구성되어 대역폭이 좁아 해당 시스템의 스펙트럼 효율을 높이는 방법은 자기 유도 통신의 매우 중요한 이슈 중 하나이다. 본 논문에서는 이러한 문제점을 해결하기 위해 대역폭 자원을 효율적으로 사용하는 OFDM 전송 방식을 고려하여 범용 소프트웨어 정의 라디오 모듈과 GNU Radio를 이용한 자기 유도 OFDM 통신 시스템을 구현하고 이의 성능을 공중에서의 실험을 통해 검증하였다.

Effect of melatonin versus injectable platelet rich fibrin on critical wound healing in submandibular salivary glands of diabetic rats

BackgroundDiabetes mellitus (DM) has harmful effects on body organs, including submandibular salivary glands (SMGs). It impairs wound healing process that follow sialoadenectomy. Yet there is no complete cure to diabetes, the available medications tend to control the side effects of DM or manage insulin resistance. Herein we tried to investigate the possible effects of injectable platelet rich fibrin (i-PRF) and melatonin on wound healing in diabetic rats. Material and methodsSurgical defects were created in SMGs of 30 rats after confirmation of DM induction. Then rats were randomly and equally allocated into three groups. Group I served as control group; group II received topically applied i-PRF, and group III received topically administrated melatonin. After 28 days all rats were euthanized, and SMGs were processed for histological and biochemical analysis. ResultsBoth i-PRF and melatonin caused significant reduction of malondialdehyde (P < 0.0001) and caspase-3 (P < 0.001) and significant increase in vascular endothelial growth factors (P = 0.001,0.009 respectively) that increased SMGs regenerative capacity when compared to diabetic group. Melatonin showed superior results regarding the histomorphological structure of SMGs. ConclusionMelatonin and i-PRF can be possible candidates for improvement of wound healing events in SMGs of diabetic rats.

Modulation of Gut Microbiota Profile and Short-Chain Fatty Acids of Rats Fed with Taro Flour or Taro Starch.

To investigate the effect of flour and starch of the Indonesian native tuber “taro” on the composition and activity of the gut microbiota in diabetic rats, streptozotocin (STZ)-induced diabetic rats were fed normal chow (AIN), or AIN in which corn starch was replaced by either taro flour or purified taro starch for 4 weeks. Fecal samples were collected at baseline and after 4 weeks, and the composition of microbial communities was measured using 16S rRNA sequencing, while SCFAs were measured using ion chromatography. Bodyweight declined upon DM induction with STZ. Feeding taro starch led to a lower reduction in bodyweight than feeding taro starch, but this was only significant for taro starch in weeks 2, 3, and 4 (p = 0.02, p = 0.01, and p < 0.01, respectively). Both taro starch and taro flour induced changes in the gut microbiota composition compared to AIN, which were different for taro flour and taro starch. Bifidobacterium, Sutterella, and Prevotella were markers for taro flour feeding, while Anaerostipes was a marker for taro starch feeding. Induction of diabetes also led to changes in the microbiota composition. Random Forest correctly predicted for 16 of 18 samples whether rats were diabetic or not and correctly predicted 6 of 12 microbiota samples belonging to either taro flour- or taro starch-fed groups, indicating also some significant overlap in the substrate, as expected. Taro starch and taro flour both led to a significant increase in the fecal concentrations of acetate, propionate, and butyrate.

Aqueous extract from Mangifera indica Linn. (Anacardiaceae) leaves exerts long-term hypoglycemic effect, increases insulin sensitivity and plasma insulin levels on diabetic Wistar rats.

BackgroundDiabetes mellitus is one of the most common todays public health problems. According to a survey by the World Health Organization, this metabolic disorder has reached global epidemic proportions, with a worldwide prevalence of 8.5% in the adult population.ObjectivesThe present study aimed to investigate the hypoglycemic effect of aqueous extract of Mangifera indica (EAMI) leaves in streptozotocin-induced diabetic rats.MethodsSixty male rats were divided into 2 groups: Normoglycemic and Diabetic. Each group was subdivided into negative control, glibenclamide 3 or 10 mg/kg, EAMI 125, 250, 500, and 1000 mg/kg. Intraperitoneal injection of streptozotocin 100 mg/kg was used to DM induction. The hypoglycemic response was assessed acutely after two and four weeks of treatment. After a 6-hour fasting period, the fasting blood glucose of animals was verified, and 2.5 g/kg glucose solution was orally administered. The insulin tolerance test and plasma insulin levels assessment were performed in the morning after fasting of 12 to 14 hours.Results and conclusionThe chemical analysis of EAMI showed high levels of phenolic compounds. There was no significant difference in fasting blood glucose between normoglycemic and diabetic groups, and that EAMI did not have an acute effect on diabetes. After two and four weeks of treatment, the extract significantly reduced blood glucose levels, exceeding glibenclamide effects. EAMI was effective in maintaining the long-term hypoglycemic effect, as well as, significantly increased the sensitivity of diabetic animals to insulin and the plasma insulin level.

Experimental induction of DM in rats and the role of bariatric surgery for the recovery of pancreatic β cells

Experimental induction of DM in rats and the role of bariatric surgery for the recovery of pancreatic β cells

EFFECTIVE DOSE OF STREPTOZOTOCIN FOR INDUCTION OF DIABETES MELLITUS AND ASSOCIATED MORTALITY RATE IN WISTAR ALBINO RATS

Background: To understand severity and complications of diabetes mellitus and to analyze effects of drugs, it is necessary to create diabetic animal model. There are different doses of streptozotocin to induce diabetes mellitus in rats that may be associated with mortality or may be insufficient for induction of DM. The objective of our study was to optimize the dose of streptozotocin to create a diabetic animal model with sustained hyperglycemia and to document the toxic dose at which there may be high mortality rate. Methods: This experimental animal study was conducted at animal house, faculty of pharmacy Ziauddin University Karachi in April 2019. The sample size included 30 albino wistar rats divided into five Groups A, B, C, D and E “with 6 rats each group”. Group A was the control, while streptozotocin at different concentrations was administered intraperitoneally in Group B, C, D and E respectively. Blood sample was drawn from lateral tail vein of animals and hyperglycemic profile was checked on 2nd, 4th, 6th, 8th and 10th day. Results: When compared to control Group A, hyperglycemic profile (blood glucose level &gt;180) was achieved in Group B, C, D and E after 48 hours. High mortality rate was observed in Group E followed by Group D. Group C had persistent hyperglycemia while Group B had reversible hyperglycemic profile. Conclusion: Intraperitoneal dose of streptozotocin 60 mg/kg created diabetic animal model with persistent hyperglycemia. However, dose above increased the mortality rate and below failed to create diabetic animal model.

Crocin treatment decreased pancreatic atrophy, LOX-1 and RAGE mRNA expression of pancreas tissue in cholesterol-fed and streptozotocin-induced diabetic rats.

Objective Oxidative stress in diabetic mellitus is a consequence of oxidative stress, which plays a critical role in the pathogenesis of diabetic tissue damage. Receptors for advanced glycation end products and for oxidized low-density lipoproteins (LDL) have critical contribution in oxidative tissue damage. The present study investigated whether anti-diabetic effects of Crocin via modulation of mRNA expression of RAGE and LOX-1 receptors in diabetic rats. Methods In the current study, high-fat cholesterol (HFC) and streptozotocin (40 mg/kg) used to induce type II diabetes. Experimental groups as follows: (Group 1: control); (Group 2: control treatment [Crocin]); (Group 3: DM [STZ]); (Group 4: DM treatment [STZ + Crocin]); (Group 5; DM + HFC [STZ + HFC]); (Group 6; DM + HFC treatment [STZ + HFC + Crocin]). Crocin (20 mg/kg/day, i.p.) administered in treatment groups for 60 days. Serum glucose and cholesterol levels evaluated on days 5, 30 and 60 after induction of DM. Pancreatic tissue from all group removed on day 60 for histological and RT-PCR analysis. Results Application of Crocin significantly decreased serum cholesterol levels on day 60 after induction of DM in diabetic + HFC rats. Moreover, Crocin significantly decreased serum glucose levels on days 30 and 60 both in diabetic and diabetic + HFC rats. Crocin partially prevented the atrophic effects of STZ on both exocrine and endocrine parts of pancreas. Additionally, Crocin significantly decreased LOX-1 and RAGE mRNA expression OF pancreas in diabetic rats. Conclusion The current study suggested that Crocin suppressed atrophic change of the pancreas by decrease of LOX-1 and RAGE mRNA expression in diabetic rats.

2479-PUB: Mesenchymal Stromal Cells Improves Motor Function and Increases Collagen Macromolecular Organization in Muscles of Diabetic Rats

Diabetes mellitus affects the skeletal muscle connective tissue and leads to significant physical performance impairment. The use of mesenchymal stromal cells (MSCs) is an attractive strategy to modify the microenvironment and promote tissue repair and regeneration. Male Wistar rats (225 ± 25 g) were randomly assigned to 3 groups: Control (C), Diabetic (DM) and Diabetic treated with MSCs (DM-MSCs). Diabetes was induced by Streptozotocin (50 µg/kg). Bone marrow cells were isolated from the tibia and femur. After 10 weeks of DM induction, DM-MSC rats received 4 i.p. injections of MSCs (1 x 106). Ten weeks after MSC transplantation motor performance was evaluated by Rota Rod test and the anterior tibial (TA) muscles were collected for morphometric and quantification of collagen birefringence by polarizing microscopy analysis. Statistical significance was set at 0.05. Fasting glycemia was significantly higher and body mass was significantly lower in the Diabetic (DM) and Diabetic treated with MSCs (DM + MSCs) groups compared with control group. Motor performance of the DM group (30.1 ± 9.6 s) was significantly reduced when compared to the C group (727.2 ± 92 s) and increased significantly in DM + MSCs group (786.8 ± 109.5 s). The TA muscle mass was significantly reduced in DM and DM + MSCs groups compared with C group. The muscle fiber cross sectional area followed the same pattern. The connective tissue increased in DM group compared with C group and decreased in DM + MSCs. The percentage collagen birefringence quantified in the connective tissue of the TA muscle decreased significantly in DM group when compared to C group and increased in DM + MSCs group. Motor performance was positively correlated with collagen birefringence (r = 0.83; P &amp;lt; 0.0001) and negatively correlated with percentage of connective tissue (r = -0.94; P &amp;lt; 0.0001). The results indicate that improves both motor function and the collagen macromolecular organization in type 1 DM. Disclosure G.L. Luna: None. M.A. Sabadine: None. Y. Estrada-Bonilla: None. F. Freitas Aníbal: None. Â.M. Leal: None. Funding Conselho Nacional de Desenvolvimen to Científico e Tecnológico Processo (447518/2014-3)

Correction: Type-2 diabetic aldehyde dehydrogenase 2 mutant mice (ALDH 2*2) exhibiting heart failure with preserved ejection fraction phenotype can be determined by exercise stress echocardiography.

[This corrects the article DOI: 10.1371/journal.pone.0195796.].

Diabetes and High-Fat Diet Induce Different Pathologies in Mouse Skeletal Muscle Extracellular Matrix

Whether obesity and diabetes (DM) similarly affect composition and function of skeletal muscle extracellular matrix (ECM) has not been resolved. This study examines the effects of long term high fat diet (HFD) and DM on rodent skeletal muscle ECM were addressed. Five week old C57BL/6 male mice were randomly allocated to standard chow (control; n=26) or HFD (45% kcal fat; n=25). After 15 weeks chow and HFD mice were rendered diabetic (by low dose STZ, 65mg/kgx3d, I.P.) (DM, n=14; HFD+DM, n=14); nondiabetic mice acted as controls. Mice were euthanased after 30 weeks and plasma and quadriceps muscle were collected for analysis. Body weight and plasma insulin expectedly decreased with DM induction and increased with HFD (all p&amp;lt;0.05). High glucose occurred only in DM mice (∼14 mM; p&amp;lt;0.05), whereas only HFD increased muscle triglyceride (p&amp;lt;0.05). Both conditions induced a decrease in quadriceps weight and in grip strength performance (p&amp;lt;0.05). Quadriceps of DM mice showed increased CTGF/CCN2 (1.5-fold; p&amp;lt;0.05), TIMP-1 (2-fold; p&amp;lt;0.05), collagen-I (3-fold; p&amp;lt;0.05), -IV (2-fold; p&amp;lt;0.05), and -VI (2-fold; p&amp;lt;0.05) protein. In contrast, HFD induced a downregulation of CTGF/CCN2 (0.6-fold; p&amp;lt;0.05), MMP-1 (0.6-fold; p&amp;lt;0.05), and TIMP-2 (0.4-fold; p&amp;lt;0.05). In DM mice quadriceps centralized nuclei (p&amp;lt;0.05) and caspase 3 (protein; 1.5-fold; p&amp;lt;0.05) were increased, whereas caspase 3 was decreased in HFD (0.5-fold; p&amp;lt;0.05). Moreover, HFD induced a downregulation of ubiquitin ligase atrogin-1 protein (0.5-fold; p&amp;lt;0.05) and mRNA levels of serine/threonine kinase p70S6k (0.75-fold; p&amp;lt;0.05). Notably, no greater dysregulation of the muscle changes observed with DM alone or with HFD alone, was seen in HFD+DM mice. These data suggest that DM and HFD differentially affect skeletal muscle ECM: DM promotes a pro-fibrotic profile with signs of muscle damage, and HFD impairs regulatory protein turnover pathways, each inducing muscle dysfunction. Disclosure S.F. Martinez-Huenchullan: None. L.A. Ban: None. A. Tao: None. S.S. Sutanto: None. C. Tam: None. S. McLennan: None. S.M. Twigg: Advisory Panel; Self; Abbott. Consultant; Self; Abbott. Advisory Panel; Self; Novo Nordisk Inc., Sanofi-Aventis, AstraZeneca, Boehringer Ingelheim Pharmaceuticals, Inc., Eli Lilly and Company. Speaker's Bureau; Self; AstraZeneca, Merck Sharp &amp; Dohme Corp.. Other Relationship; Self; Abbott.