scholarly journals Loss of Response Gene to Complement 32 (RGC-32) in Diabetic Mouse Retina Is Involved in Retinopathy Development

2018 ◽  
Vol 19 (11) ◽  
pp. 3629 ◽  
Author(s):  
Wen-Ling Liao ◽  
Jane-Ming Lin ◽  
Shih-Ping Liu ◽  
Shih-Yin Chen ◽  
Hui-Ju Lin ◽  
...  

Diabetic retinopathy (DR) is a severe and recurrent microvascular complication in diabetes. The multifunctional response gene to complement 32 (RGC-32) is involved in the regulation of cell cycle, proliferation, and apoptosis. To investigate the role of RGC-32 in the development of DR, we used human retinal microvascular endothelial cells under high-glucose conditions and type 2 diabetes (T2D) mice (+Leprdb/ + Leprdb, db/db). The results showed that RGC-32 expression increased moderately in human retinal endothelial cells under hyperglycemic conditions. Histopathology and RGC-32 expression showed no significant changes between T2D and control mice retina at 16 and 24 weeks of age. However, RGC-32 expression was significantly decreased in T2D mouse retina compared to the control group at 32 weeks of age, which develop features of the early clinical stages of DR, namely reduced retinal thickness and increased ganglion cell death. Moreover, immunohistochemistry showed that RGC-32 was predominantly expressed in the photoreceptor inner segments of control mice, while the expression was dramatically lowered in the T2D retinas. Furthermore, we found that the level of anti-apoptotic protein Bcl-2 was decreased (approximately 2-fold) with a concomitant increase in cleaved caspase-3 (approximately 3-fold) in T2D retina compared to control. In summary, RGC-32 may lose its expression in T2D retina with features of DR, suggesting that it plays a critical role in DR pathogenesis.

2020 ◽  
pp. 1-14
Author(s):  
Yaser H.A. Elewa ◽  
Osamu Ichii ◽  
Teppei Nakamura ◽  
Yasuhiro Kon

Diabetes is a devastating global health problem and is considered a predisposing factor for lung injury progression. Furthermore, previous reports of the authors revealed the role of mediastinal fat-associated lymphoid clusters (MFALCs) in advancing respiratory diseases. However, no reports concerning the role of MFALCs on the development of lung injury in diabetes have been published. Therefore, this study aimed to examine the correlations between diabetes and the development of MFALCs and the progression of lung injury in a streptozotocin-induced diabetic mouse model. Furthermore, immunohistochemical analysis for immune cells (CD3+ T-lymphocytes, B220+ B-lymphocytes, Iba1+ macrophages, and Gr1+ granulocytes), vessels markers (CD31+ endothelial cells and LYVE-1+ lymphatic vessels “LVs”), and inflammatory markers (TNF-α and IL-5) was performed. In comparison to the control group, the diabetic group showed lung injury development with a significant increase in MFALC size, immune cells, LVs, and inflammatory marker, and a considerable decrease of CD31+ endothelial cells in both lung and MFALCs was observed. Furthermore, the blood glucose level showed significant positive correlations with MFALCs size, lung injury, immune cells, inflammatory markers, and LYVE-1+ LVs in lungs and MFALCs. Thus, we suggest that the development of MFALCs and LVs could contribute to lung injury progression in diabetic conditions.


2018 ◽  
Vol 15 (2S) ◽  
pp. 145-152
Author(s):  
S. V. Shukhaev ◽  
E. V. Boiko

Purpose: to compare two types of phacoemulsification parameters (combination ultrasound and torsional US with IP) with estimating the number of postoperative complications caused by intraoperative trauma of corneal endothelial cells.Patients and methods. 72 patients underwent phacoemulsification with IOL implantation. Patients were randomly divided into two groups (main n = 33 and control n = 39). During the aspiration of lens fragments in the main group the combination ultrasound was used, while torsional US with IP was used in the control group. Endothelial cell counting and other examinations were performed 1 day, 1 week and 6 months after the surgery.Results. CDVA in the explored groups 1 week after the surgery was similar: the main group — 0.813 ± 0.228, the control group — 0.765 ± 0.250, There was also no statistically significant difference in the thickness of the cornea between the groups: the main group — 533.48 ± 12.41, the control group — 536.44 ± 10.92. At the same time, a statistically significant difference was found in the density of endothelial cells: the main group: 1871.30 ± 187.41 (after 1 week), 1865 ± 178.9 (after 6 months); control group: 1809.63 ± 225.43 (after 1 week), 1791 ± 230.82 (after 6 months). The percentage of cell loss was, respectively, lower in the main group at all times of observation: 3.90% (after 1 day), 4.54% (after 1 week), 4.9% (after 6 months). In the control group: 7.71% (after 1 day), 9.25% (after 1 week), 10.4% (after 6 months).Conclusions. Data obtained in this study has showed the advantages of combination ultrasound in comparison with torsional US with IP, when performing aspiration of dense lenses. Due to lower consumption of ultrasound energy and higher aspiration rate of the fragments, combination ultrasound can reduce the loss of corneal endothelial cells and decrease the number and severity of postoperative complications associated with it. 


Author(s):  
Namita Chatterjee ◽  
Eugenia Fraile-Bethencourt ◽  
Adrian Baris ◽  
Cristina Espinosa-Diez ◽  
Sudarshan Anand

Defects in stress responses are important contributors in many chronic conditions including cancer, cardiovascular disease, diabetes, and obesity-driven pathologies like non-alcoholic steatohepatitis (NASH). Specifically, endoplasmic reticulum (ER) stress is linked with these pathologies and control of ER stress can ameliorate tissue damage. MicroRNAs have a critical role in regulating diverse stress responses including ER stress. Here, we show that miR-494 plays a functional role during ER stress. Pharmacological ER stress inducers (tunicamycin (TCN) and thapsigargin) and hyperglycemia robustly increase the expression of miR-494 in vitro. ATF6 impacts the primary miR-494 levels whereas all three ER stress pathways are necessary for the increase in mature miR-494. Surprisingly, miR-494 pretreatment dampens the induction and magnitude of ER stress in response to TCN in endothelial cells and increases cell viability. Conversely, inhibition of miR-494 increases ER stress de novo and amplifies the effects of ER stress inducers. Using Mass Spectrometry (TMT-MS) we identified 23 proteins that are downregulated by both TCN and miR-494 in cultured human umbilical vein endothelial cells. Among these, we found 6 transcripts which harbor a putative miR-494 binding site. We validated the anti-apoptotic gene BIRC5 (survivin) and GINS4 as targets of miR-494 during ER stress. In summary, our data indicates that ER stress driven miR-494 may act in a feedback inhibitory loop to dampen downstream ER stress signaling.


2020 ◽  
Vol 1 (1) ◽  
pp. 37-41
Author(s):  
Ting-Ting Zhao ◽  
◽  
Jing Jin ◽  
Shu-Nong Xie ◽  
Ying Fan ◽  
...  

AIM: To investigate the relationship of matrix metalloproteinase-2 (MMP-2) and transforming growth factor-beta 2 (TGF-β2) levels in human vitreous with axial length (AL) of patients with high myopia. METHODS: The concentrations of MMP-2 and TGF-β2 levels were tested by enzyme-linked immunosorbnent assay (ELISA). Fifty-five human vitreous samples of 55 patients were collected during vitrectomy surgery, and were divided into two groups according to their spherical equivalent (SE) and AL. High myopia group (25 cases): AL≥26.00 mm, and control group or non-high myopia group (30 cases): AL<26.00 mm. RESULTS: The MMP-2 levels in vitreous of high myopia group (96.87±55.95 ng/mL) was significantly higher than that of control group (77.24±41.81 ng/mL, P<0.05), but not correlated with AL (r=0.088, P=0.544). While the vitreous TGF-β2 concentration was negatively correlated with AL (r=-0.344, P=0.014), and there was significant difference of TGF-β2 vitreous levels between high myopia group (3729.08±1890.88 pg/mL) and control group (3926.00± 1333.88 pg/mL, P<0.05). CONCLUSION: MMP-2 and TGF-β2 in human vitreous may play a critical role in human high myopia development, and the TGF-β2 appears to be associated with AL.


2021 ◽  
Author(s):  
Elena Ivanova ◽  
Carlo Corona ◽  
Cyril G. Eleftheriou ◽  
Randy F. Stout ◽  
Jakob Korbelin ◽  
...  

Endothelial cells (ECs) are key players in the development and maintenance of the vascular tree, the establishment of the blood brain barrier and control of blood flow. Disruption in ECs is an early and active component of vascular pathogenesis. However, our ability to selectively target ECs in the CNS for identification and manipulation is limited. Here, in the mouse retina, a tractable model of the CNS, we utilized a recently developed AAV-BR1 system to identify distinct classes of ECs along the vascular tree using a GFP reporter. We then developed an inducible EC-specific ectopic Connexin 43 (Cx43) expression system using AAV-BR1-CAG-DIO-Cx43-P2A-DsRed2 in combination with a mouse line carrying inducible CreERT2 in ECs. We targeted Cx43 because its loss has been implicated in microvascular impariment in numerous diseases such as diabetic retinopathy and vascular edema. GFP-labeled ECs were numerous, evenly distributed along the vascular tree and their morphology was polarized with respect to the direction of blood flow. After tamoxifen induction, ectopic Cx43 was specifically expressed in ECs. Similarly to endogenous Cx43, ectopic Cx43 was localized at the membrane contacts of ECs and it did not affect tight junction proteins. The ability to enhance gap junctions in ECs provides a precise and potentially powerful tool to treat microcirculation deficits, an early pathology in numerous diseases.


2021 ◽  
Vol 11 (12) ◽  
pp. 2427-2434
Author(s):  
Zexue Zhao ◽  
Pengfei Wu ◽  
Jiwei Tian ◽  
Yifan Yu

Our study assessed the role of Wnt signaling inhibitor (SM04690) in fracture healing and the underlying mechanism. Sprague Dawley (SD) rats were used to establish a fracture model which was then separated into SM04690 group which received SM04690 (50 mg/kg) by intraperitoneal injection once a day for one week, and control group which received saline once a day. After rats were sacrificed, the fractured femurs were harvested to measure femoral strength by stress testing, bone density and volume by CT. Femurs were sliced for immunohistochemical staining. Mesenchymal stem cells (MSCs), endothelial cells, osteoprogenitor cells and osteoblasts were detected by flow cytometer and EFNB1 expression was detected by immunoblotting and PCR. In addition, MSCs were treated with SM04690 (5 uM), followed by detection of EFNB1 expression. SM04690 treatment significantly inhibited EFNB1 expression and reduced bone volume and callus volume as well as decreased ultimate load of bones. Immunohistochemical staining and flow cytometry analysis showed no difference of osteoclast numbers at the fracture site between two groups, but proportion of osteoclasts in the cartilage tissue of SM04690 group was significantly decreased. In addition, the number of osteoblasts, osteoprogenitor cells and endothelial cells was significantly decreased after treatment. Under the conditions favoring osteogenic differentiation, the production of minerals by osteogenic cells was significantly decreased along with upregulated TAZ phosphorylation and downregulated Osterix in SM04690 group. In conclusion, SM04690 delays fracture healing by inhibiting EpRunB1 in osteoprogenitor cells.


2018 ◽  
Vol 100 ◽  
Author(s):  
POLINA V. POPOVA ◽  
LIUDMILA B. VASILEVA ◽  
ALEXANDRA S. TKACHUK ◽  
MAXIM V. PUZANOV ◽  
YANA A. BOLOTKO ◽  
...  

SummaryMaternal gestational diabetes mellitus (GDM) is considered to be an important factor that epigenetically predisposes offspring to metabolic and cardiovascular diseases. However, the mechanisms of how intrauterine hyperglycaemia affects offspring have not been thoroughly studied. The mammalian tribbles homologue 1 (TRIB1) gene is associated with plasma lipid concentrations and coronary artery disease (CAD). Our aim was to study the effect of GDM and its treatment terms on the level of TRIB1 gene expression in human umbilical vein endothelial cells (HUVECs) of newborns from women with and without GDM. The study included 50 women with GDM and 25 women without GDM (control group). Women with GDM were divided into three groups according to their gestational age when the treatment of GDM started: 24–28 weeks (GDM1, N = 16), 29–32 weeks (GDM2, N = 25) and >34 weeks (GDM3, N = 9). The levels of TRIB1 gene expression in GDM3, GDM2, GDM1 and control groups were 2.8 ± 1.1, 4.2 ± 2.4, 6.0 ± 3.4 and 8.1 ± 6.1, respectively (p = 0.001). After comparison in pairs the difference was significant for the following pairs: GDM2–control (p = 0.004), GDM3–control (p = 0.002), GDM1–GDM3 (p = 0.012). Notably, if treatment had been started before the 28th week of gestation, the difference in TRIB1 gene expression in HUVECs was not significant (p = 0.320 for comparison between GDM1 and control groups). Our findings support the hypothesis that TRIB1 gene expression in HUVECs depends on the duration of intrauterine exposure to hyperglycaemia.


2010 ◽  
Vol 80 (1) ◽  
pp. 65-73 ◽  
Author(s):  
Pei-Min Chao ◽  
Wan-Hsuan Chen ◽  
Chun-Huei Liao ◽  
Huey-Mei Shaw

Conjugated linoleic acid (CLA) is a collective term for the positional and geometric isomers of a conjugated diene of linoleic acid (C18:2, n-6). The aims of the present study were to evaluate whether levels of hepatic α-tocopherol, α-tocopherol transfer protein (α-TTP), and antioxidant enzymes in mice were affected by a CLA-supplemented diet. C57BL/6 J mice were divided into the CLA and control groups, which were fed, respectively, a 5 % fat diet with or without 1 g/100 g of CLA (1:1 mixture of cis-9, trans-11 and trans-10, cis-12) for four weeks. α-Tocopherol levels in plasma and liver were significantly higher in the CLA group than in the control group. Liver α-TTP levels were also significantly increased in the CLA group, the α-TTP/β-actin ratio being 2.5-fold higher than that in control mice (p<0.01). Thiobarbituric acid-reactive substances were significantly decreased in the CLA group (p<0.01). There were no significant differences between the two groups in levels of three antioxidant enzymes (superoxide dismutase, glutathione peroxidase, and catalase). The accumulation of liver α-tocopherol seen with the CLA diet can be attributed to the antioxidant potential of CLA and the ability of α-TTP induction. The lack of changes in antioxidant enzyme protein levels and the reduced lipid peroxidation in the liver of CLA mice are due to α-tocopherol accumulation.


Author(s):  
Abdullah Farih

The objective of this study is to measure whether there is or not significant effect of teaching reading by utilizing Hot Potatoes software toward students’ reading comprehension of descriptive text. The variables used were the teaching treatment of Hot Potatoes Software as the independent variable and the students’ reading comprehension of Descriptive Text as dependent variable. Then, the research designed was Quasi-experimental design and the presentation of data used was quantitative. The data were obtained from the eleventh grade, of which 28 students were taken as sample. The students are divided into two groups; experiment group consisted of 12 students and control group consists of 16 students. To get the data, the pre-test and post-test were applied and then analyzed using t-test formula. The researcher had concluded that there is Significant effect of Hot Potatoes Software toward the students’ reading comprehension achievement. The result showed that mean of students’ post-test was increased. It is proved by the t-test (8.54) which is higher than t-table (2.05) at level of significance 5%. It means that the alternative hypothesis was accepted and it proved that Hot Potatoes Software had significant effect toward students’ reading comprehension of Descriptive Text


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