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BackgroundType 1 diabetes is caused by a chronic immune response that destroys islet beta cells， resulting in elevated blood glucose. Mesenchymal stem cells can prevent and treat the development of diabetes and its complications. However， little is known about the effects and potential mechanisms of Gingival mesenchymal stem cells （GMSCs） in preventing diabetes. The aim of this study is to investigate the mechanism of GMSCs in preventing type 1 diabetes in mice and to find targets for clinical treatment of diabetes. MethodsWe injected human GMSCs into NOD mice to observe the trend of blood glucose， observed the survival of pancreatic β-cells by immunohistochemistry， and detected the change of immune cells in the spleen of mice by flow analysis. Finally， the immune cells in NOD mice were transfused into NOD-SCID mice to observe the onset of diabetes in NOD-SCID mice. ResultsGMSCs significantly reduced the incidence of diabetes in NOD mice， with 64% of control mice developing diabetes at 27 weeks of age compared with 35% in the GMSC group， P=0.013. The percentage of Follicular B cells（FO B cell） in the spleen of GMSCs-treated mice decreased from （52.2±4.1）% to （43.2±5.3）%， P=0.008， while other types of immune cells did not change significantly. The immunohistochemical results showed that GMSCs could effectively improve the survival of pancreatic β-cells， which could continuously produce insulin to control blood glucose. Finally， we found the spleen cells transfusion could prevent the development of diabetes in NOD-SCID mice. ConclusionGMSCs can reduce diabetes in mice by reducing FO B cells in the spleen.
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@#AIM:To determine the pathological changes in ocular surfaces dry eye excessive evaporation non-obese diabetic(NOD)mice model and to preliminarily explore the feasibility of diabetic dry eye model.<p>METHODS: In this study, 40 females NOD mice were selected. The experimental group consisted of NOD mice that were diagnosed with diabetes while the normal control group consisted of those NOD mice without spontaneous diabetes. Hypodermic injection of Scopolamine hydrobromide(0.5mg/0.2mL)was administered under 40% humidity to the experimental group and placed in a controlled drying box for 12h a day. This was to achieve a dry eye model. Testing indicators on the 1, 7, 10 and 14d after modeling, phenol red thread test was used to measure tear secretion and the eye sections were stained with periodic acid-Schiff(PAS)to examine the morphology and number of conjunctival goblet cells. On the 10d after modeling, the changes in the corneal epithelium were visualized after staining with hematoxylin. <p>RESULTS:For the NOD mice of the experimental group, the tear secretion was gradually decreased with timing, while there were no obvious changes in the normal control group. The volume of the conjunctival goblet cells of the experimental group became larger, and on the 1d after the molding, the experimental group had decreased density of the goblet cells when compared with the normal control group(<i>P</i>=0.008). From the 7d after the molding, as the time was prolonged, the density of the goblet cells was gradually decreased and the differences between the two group at same time point were significant(all <i>P</i><0.001). Besides, it was required to observe the corneal epithelium of the two groups on the 10d. The result shows that the corneal epithelium became thinned, some epithelial cells were denatured, and stromal cells became edema.<p>CONCLUSION: Dry eye model of NOD mice was preliminary established, and the changes of ocular surface were similar to those of dry eye in the clinic.
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Objective To explore the effects of DPP-4 inhibitor sitagliptin on insulitis of NOD mice and the potential mechanism. Methods 223-month-old female NOD mice were randomly divided into Sitagliptin(Si-ta)group(n=11)gavaged with Sita(30 mg/Kg)daily for 12 weeks and control(Con)group with equal volume of normal saline(NS). The body weight,food-intake,water-intake and blood glucose were recorded weekly. Intraperi-toneal glucose tolerance test(IPGTT)was performed at the end of treatment and the blood sample was collected. Then mice were execute. The serum insulin level was measured by ELISA. Pancreas morphology and insulitis were evaluated by hematoxylin-eosin staining. The protein expression level of TLR4 ,MyD88 and NF-κB was analyzed by Western Blot. Results Compared with that of Con group ,the insulitis of Sita group was significantly alleviative(P < 0.05)and the serum insulin level was increased significantly(P < 0.05). The protein expression level of TLR4(P<0.05),MyD88(P<0.01)and NF-κB(P<0.05)in Sita group was significantly decreased. Conclusion Sitagliptin alleviates insulitis in NOD mice and increases the serum insulin level ,probably owing to the suppression of TLR4/MyD88/NF-κB signaling pathway.
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Objective To explore the effects of DPP-4 inhibitor sitagliptin on insulitis of NOD mice and the potential mechanism. Methods 223-month-old female NOD mice were randomly divided into Sitagliptin(Si-ta)group(n=11)gavaged with Sita(30 mg/Kg)daily for 12 weeks and control(Con)group with equal volume of normal saline(NS). The body weight,food-intake,water-intake and blood glucose were recorded weekly. Intraperi-toneal glucose tolerance test(IPGTT)was performed at the end of treatment and the blood sample was collected. Then mice were execute. The serum insulin level was measured by ELISA. Pancreas morphology and insulitis were evaluated by hematoxylin-eosin staining. The protein expression level of TLR4 ,MyD88 and NF-κB was analyzed by Western Blot. Results Compared with that of Con group ,the insulitis of Sita group was significantly alleviative(P < 0.05)and the serum insulin level was increased significantly(P < 0.05). The protein expression level of TLR4(P<0.05),MyD88(P<0.01)and NF-κB(P<0.05)in Sita group was significantly decreased. Conclusion Sitagliptin alleviates insulitis in NOD mice and increases the serum insulin level ,probably owing to the suppression of TLR4/MyD88/NF-κB signaling pathway.
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Background In Brazil, accidents with venomous animals are considered a public health problem. Tityus serrulatus (Ts), popularly known as the yellow scorpion, is most frequently responsible for the severe accidents in the country. Ts envenoming can cause several signs and symptoms classified according to their clinical manifestations as mild, moderate or severe. Furthermore, the victims usually present biochemical alterations, including hyperglycemia. Nevertheless, Ts envenoming and its induced hyperglycemia were never studied or documented in a patient with diabetes mellitus (DM). Therefore, this is the first study to evaluate the glycemia during Ts envenoming using a diabetic animal model (NOD, non-obese diabetic). Methods Female mice (BALB/c or NOD) were challenged with a non-lethal dose of Ts venom. Blood glucose level was measured (tail blood using a glucose meter) over a 24-h period. The total glycosylated hemoglobin (HbA1c) levels were measured 30 days after Ts venom injection. Moreover, the insulin levels were analyzed at the glycemia peak. Results The results demonstrated that the envenomed NOD animals presented a significant increase of glycemia, glycosylated hemoglobin (HbA1c) and insulin levels compared to the envenomed BALB/c control group, corroborating that DM victims present great risk of developing severe envenoming. Moreover, the envenomed NOD animals presented highest risk of death and sequelae. Conclusions This study demonstrated that the diabetic victims stung by Ts scorpion should be always considered a risk group for scorpion envenoming severity.(AU)
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Animals , Mice , Scorpions , Glycated Hemoglobin , Public Health , Risk Factors , Scorpion Stings , Diabetes MellitusABSTRACT
In Brazil, accidents with venomous animals are considered a public health problem. Tityus serrulatus (Ts), popularly known as the yellow scorpion, is most frequently responsible for the severe accidents in the country. Ts envenoming can cause several signs and symptoms classified according to their clinical manifestations as mild, moderate or severe. Furthermore, the victims usually present biochemical alterations, including hyperglycemia. Nevertheless, Ts envenoming and its induced hyperglycemia were never studied or documented in a patient with diabetes mellitus (DM). Therefore, this is the first study to evaluate the glycemia during Ts envenoming using a diabetic animal model (NOD, non-obese diabetic). Methods Female mice (BALB/c or NOD) were challenged with a non-lethal dose of Ts venom. Blood glucose level was measured (tail blood using a glucose meter) over a 24-h period. The total glycosylated hemoglobin (HbA1c) levels were measured 30 days after Ts venom injection. Moreover, the insulin levels were analyzed at the glycemia peak. Results The results demonstrated that the envenomed NOD animals presented a significant increase of glycemia, glycosylated hemoglobin (HbA1c) and insulin levels compared to the envenomed BALB/c control group, corroborating that DM victims present great risk of developing severe envenoming. Moreover, the envenomed NOD animals presented highest risk of death and sequelae. Conclusions This study demonstrated that the diabetic victims stung by Ts scorpion should be always considered a risk group for scorpion envenoming severity.
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Animals , Rats , Poisoning/complications , Poisoning/mortality , Rats/abnormalities , Scorpions/chemistryABSTRACT
Objective:To illuminate the influence of InsB15-23 H-2Kd dtSCT to the morbidity of type 1 diabetes mellitus in NOD mice.Methods:An eukaryotic plasmid encoded membrane-expressed InsB15-23 H-2Kd dtSCT was inoculated into 3 weeks old female NOD mice subcutaneously and the blood sugar and morbidity of type 1 diabetes mellitus were monitored once a week.To illuminate the cellular mechanism of immunologic intervention of membrane-expressed InsB15-23 H-2Kd dtSCT to the course of type 1 diabetes mellitus in NOD mice,the mononuclear cell infiltration of islets was detected by tissue slice and the frequency of IGRP206 2-14 specific CTLs in PBMC was analyzed by FACs.Results: As compared with pcDNA3.1 (-) control ( 60%) and untreated NOD mice ( 80%) , mice immunized with InsB15-23 H-2Kd dtSCT exhibited low level of islet infiltration and low morbidity in 30 weeks old ( 9%) .But the frequency of IGRP206-214 specific CTLs in PBMC of 16 and 40 weeks old mice showed no difference.Conclusion:Membrane-expressed InsB15-23 H-2Kd dtSCT can protect NOD mice from type 1 diabetes mellitus in IGRP206-214 independent pattern.
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Three factors ( Oct4,Sox2,and Klf4) were introduced into tail tip-derived fibroblasts of NOD mouse by retrovirus infection.The induced pluripotent stem cells ( iPSCs ) generated from this method were analyzed in several aspects,including surface antigens,gene expression,alkaline phosphatase activity,and teratoma formation assays.The NOD mouse iPSCs generated from this study had ES-like characters,expressed ES-specific surface antigens,and possessed the ability to differentiate into 3-germ layers.Such NOD mouse-iPSCs should be useful in tyre 1 dialectic disease modeling,as well as for cell replacement therapy.
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AIM:To investigate retinal vascular endothelial growth factor(VEGF)level and retinal cells apoptosis in the early stage of diabetic NOD mouse retina.METHODS:Animals were divided into the control group(non-diabetes mice)(2,4,6,8,12 weeks group,n=30)and diabetes group(2,4,6,8,12 weeks group,n=30).EUSA(Enzyme-Linked Immunosorbent Assay)was performed to detect VEGF level in both serum and retina.Transmission electron microscope method was used to examine retinal cell apoptosis.RESULTS:Compared with the control group,VEGF levels in serum and retina were increased significantly in the NOD group(12 weeks:4.9±0.4μg/g versus 0.19±0.1μg/g in serum sample,P ＜ 0.01;165.0±9.0μg/g versus 18.0±4.0μg/g in retinal sample,P＜0.01).There exists a positive correlation between serum VEGF and retinal VEGF levels in the early diabetic NOD mice(γ=0.9902,P=0.001).The number of the cells apoptosis in the ganglion significantly in the NOD group(P＜0.01).CONCLUSION:The high VEGF expression may be contributed to increase retinal cells apoptosis.Many factors associated with retinal VEGF expression might involve in the early diabetes stage.
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O camundongo não obeso diabético (NOD) é caracterizado por desenvolver naturalmente diabetes mellitus tipo 1 (DM-1) com similaridade ao diabetes mellitus tipo 1 em humanos. A manifestação espontânea do diabetes neste modelo animal é caracterizado por infiltração progressiva das ilhotas de Langerhans por células mononucleares linfócitos T (CD4+ e CD8+) e destruição das células ß pancreáticas produtoras de insulina. O fator de crescimento neural (NGF) e algumas citocinas estão associados a regeneração neural, além de atuarem sobre células do sistema imune. Em adição a estes efeitos, NGF age na liberação de insulina pelas células betas das ilhotas pancreáticas, tornando-se foco de interesse com relação as suas propriedades moduladoras no processo inflamatório na ilhota pancreática. O gangliosídeo GM1 liga-se ao receptor de alta afinidade (TrkA) do NGF-ß, mimetizando seus efeitos. No presente trabalho, avaliamos a ação modulatória de GM1 e NGF em cultura de ilhotas pancreáticas, provenientes de camundongos NOD. Foram avaliados por meio de RT-PCR a expressão gênica de NGF-ß, TrkA e insulina e, por ensaio imunoenzimático, a concentração de citocinas IL-1ß, IL-12, TNF-a, INF-y e insulina. Nossos resultados sugerem ação moduladora similar entre GM1 e NGF sobre as ilhotas de NOD não diabéticos e pré-diabéticos. NGF e GM1 aumentam a expressão gênica de NGF e TrkA e diminuem a expressão gênica de insulina em NOD não diabéticos e pré-diabéticos. Além disso, aumentam a liberação de insulina e diminui a de citocinas inflamatórias IL-1ß, IL-12, TNF-a, IFN-y que caracterizam a resposta Th1.
The non-obese diabetic mice (NOD) lineage is characterized by developing type 1 diabetes mellitus (DM-1) naturally, bearing a similarity to DM-1 in human beings. The spontaneous manifestation of diabetes is characterized by gradual infiltration in pancreatic islets by mononuclear cells lymphocytes T (CD4+ and CD8+) and destruction of the ß-cells producers of insulin. One consequence of this effect, is the release of neurotrophins trying modulate the insulin release by the ß cells of pancreatic islets. Thus, the neurotrophins have been the focus of interest in the modulation of the inflammatory process in the pancreatic islets. The ganglioside GM1 binds to the high affinity receptor (TrkA) of the NGF-ß, enhancing its effect. In the present work, we evaluate the immune modulation properties of GM1 and NGF in culture of pancreatic islets from NOD mice. The gene expression of NGF-ß, TrkA and insulin for immune enzymatic assay, the concentration of cytokines IL 1ß, IL-12, TNF-a, IFN-y and insulin were evaluated by RT-PCR and ELISA. Our results suggest similar modulation action between GM1 and NGF on islets of NOD non-diabetic and pre-diabetic. GM1 and NGF action increases the gene expression of NGF and TrkA and the decrease of insulin in mice NOD non-diabetic and pre-diabetic. Moreover, GM1 and NGF increase the insulin release and decrease inflammatory cytokines that characterize the Th1 reply.
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Animals , Mice , Diabetes Mellitus, Type 1 , Islets of Langerhans , Mice, Inbred NOD , Transforming Growth Factors , Lymphotoxin-alpha , Receptors, Tumor Necrosis Factor , Tumor Necrosis Factor-alphaABSTRACT
Type 1 diabetes is considered as Th1 cell mediated autoimmune disease and the suppression of Th1 cells or the activation of Th2 cells has been regarded as a plausible immunologic intervention for the prevention of type 1 diabetogenesis in a rodent model. CpG ODN is an immunostimulatory sequence primarily present in bacterial DNA, viral DNA and BCG. CpG ODN is conventionally classified as a Th1 cell activator, which has been clinically applied to cancer, allergy and infectious disease. Recently, there was a promising report of that CpG ODN administration suppressed the development of type 1 diabetes in NOD mice by inducing Th2 cell mediated cytokine. However, the antidiabetogenic effect of CpG ODN on NOD mice is controversial. Thus, two studies were serially undertaken with various kinds of CpG motif to find a more optimal sequence and administration method. In the first study, CpG ODN was vaccinated four times and pancreatic inflammation and the quantity of serum insulin subsequently evaluated. In the second study, the amounts of IFN gamma and IL-4 in sera were measured as representative cytokines of Th1 and Th2 cells, respectively. As a result, vaccination or continuous injection of CpG ODN failed to show a preventive effect on type 1 diabetogenesis in NOD mice. Structural differences of CpG ODN also had no affect on the result. CpG ODN also consistently showed affect on the pancreatic pathology. The productions of IFN gamma and IL-4 were detected only in the K and D type CpG ODN administration groups. Comparison of the two cytokines leads to the conclusion that CpG ODN generated a Th1-weighted response in both study groups. It was assumed that CpG ODN failed to produce Th2-weighted cytokine milieu, which can overcome the genetically determined phenotype of NOD mice. Given these results, it was concluded that the immunotherapeutic application of CpG ODN on Type 1 diabetes had clear limitations.
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Animals , Female , Mice , DNA/pharmacology , Diabetes Mellitus, Type 1/immunology , Mice, Inbred NOD , Th1 Cells/immunologyABSTRACT
AIM To investigate the anti-inflammatory properties of IL-6 on the development of IDDM(insulin dependent diabetes mellitus) in NOD mouse. METHODS IL-6 was administered exogenously in female NOD mice (spontaneous model) and cyclophosphamide-treated NOD mice (accelerated model) respectively. During the study period, the mice were evaluated for development of diabetes once a week. At the end of experiment, 8 euglycosuria mice from each group were sacrificed for histoimmunological analysis, and the production of TNF-? and IFN-? from spleen lymphoid cells were detected. RESULTS The incidence of IDDM in spontaneous model(33%) were significantly reduced after 12 weeks of treatment. TNF-?(P
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13 2 mmol/L. Results:The incidence of diabetes in the treated group was 31 6% which was obviously less than 72 2% in control group ( P