Hyperglycemia-induced effects on glycocalyx components in the retina.
Hyperglycemia-induced effects on glycocalyx components in the retina.
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DOI：
10.1016/j.exer.2021.108846
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发表时间：
2021-12
影响因子：
3.4
通讯作者：
Harris NR
中科院分区：
文献类型：
--
作者：
Kaur G;Rogers J;Rashdan NA;Cruz-Topete D;Pattillo CB;Hartson SD;Harris NR
糖尿病视网膜病变是一种以内皮损伤和血管功能障碍为特征的危及视力的糖尿病并发症。内皮糖萼是所有内皮细胞的一个动态层，内皮糖萼的丢失会导致几种微血管病变，包括血管通透性增加、白细胞堵塞和毛细血管闭塞，并可能导致视网膜病变的进展。以前，糖萼厚度的显著减少已在糖尿病视网膜观察到。然而，糖尿病对视网膜糖萼特定成分的影响尚未被研究。因此，我们的研究目的是探讨高血糖诱导视网膜糖萼成分的合成、表达和脱落的变化，为糖尿病视网膜病变提供新的治疗靶点。原代大鼠视网膜微血管内皮细胞（RRMEC）在正常葡萄糖（5 mM）或高葡萄糖（25 mM）条件下生长6天。分别采用qRT-PCR和Western blot检测糖萼组分的mRNA和蛋白水平。此外，采用质谱法分析核心蛋白的蛋白强度。此外，采用链脲佐菌素诱导的1型糖尿病大鼠模型，研究视网膜糖萼在体内表达的变化。用Western blot方法研究了糖萼在培养基和血浆中的脱落情况。在体外和体内高糖条件下，syndecan-1和CD44的脱落均显著增加。在高糖条件下生长的RRMEC中，syndecan-3的mRNA水平显著降低，而syndecan-1、syndecan-2、syndecan-4、glypican-1、glypican-3和CD44的mRNA水平显著升高。高糖暴露后，syndecan-3和glypican-1在RRMEC中的表达显著降低，而syndecan-1和CD44的表达显著升高。此外，质谱数据还表明，在高葡萄糖刺激下，syndecan-4蛋白水平显著升高，glypican-3蛋白水平显著降低。在体内，我们的数据还表明，糖尿病大鼠视网膜中syndecan-3 mRNA转录量显著降低，glypican-1和CD44 mRNA水平显著升高。糖尿病大鼠视网膜中syndecan-3和CD44的表达显著降低。然而，syndecan-1和glypican-1在糖尿病视网膜中的表达明显增加。我们研究的主要发现之一是葡萄糖诱导的内皮糖萼各种成分的表达和脱落变化具有相当大的多样性，例如内皮和视网膜syndecan-1增加，但内皮和视网膜syndecan-3减少。这表明糖尿病视网膜糖萼的减少不是非特异性脱落机制的结果。此外，mRNA的测量也显示出类似的多样性，syndecan-1、glypican-1和CD44在内皮和/或视网膜的水平增加，但syndecan-3的水平下降，mRNA的增加可能是对糖萼整体损失的代偿反应。
Diabetic retinopathy is a vision-threatening complication of diabetes characterized by endothelial injury and vascular dysfunction. The loss of the endothelial glycocalyx, a dynamic layer lining all endothelial cells, contributes to several microvascular pathologies, including an increase in vascular permeability, leukocyte plugging, and capillary occlusion, and may drive the progression of retinopathy. Previously, a significant decrease in glycocalyx thickness has been observed in diabetic retinas. However, the effects of diabetes on specific components of the retinal glycocalyx have not yet been studied. Therefore, the aim of our study was to investigate changes in synthesis, expression, and shedding of retinal glycocalyx components induced by hyperglycemia, which could provide a novel therapeutic target for diabetic retinopathy. Primary rat retinal microvascular endothelial cells (RRMEC) were grown under normal glucose (5 mM) or high-glucose (25 mM) conditions for 6 days. The mRNA and protein levels of the glycocalyx components were examined using qRT-PCR and Western blot analysis, respectively. Further, mass spectrometry was used to analyze protein intensities of core proteins. In addition, the streptozotocin-induced Type 1 diabetic rat model was used to study changes in the expression of the retinal glycocalyx in vivo. The shedding of the glycocalyx was studied in both culture medium and in plasma using Western blot analysis. A significant increase in the shedding of syndecan-1 and CD44 was observed both in vitro and in vivo under high-glucose conditions. The mRNA levels of syndecan-3 were significantly lower in the RRMEC grown under high glucose conditions, whereas those of syndecan-1, syndecan-2, syndecan-4, glypican-1, glypican-3, and CD44 were significantly higher. The protein expression of syndecan-3 and glypican-1 in RRMEC was reduced considerably following exposure to high glucose, whereas that of syndecan-1 and CD44 increased significantly. In addition, mass spectrometry data also suggests a significant increase in syndecan-4 and a significant decrease in glypican-3 protein levels with high glucose stimulation. In vivo, our data also suggest a significant decrease in the mRNA transcripts of syndecan-3 and an increase in mRNA levels of glypican-1 and CD44 in the retinas of diabetic rats. The diabetic rats exhibited a significant reduction in the retinal expression of syndecan-3 and CD44. However, the expression of syndecan-1 and glypican-1 increased significantly in the diabetic retina. One of the main findings of our study was the considerable diversity of glucose-induced changes in expression and shedding of various components of endothelial glycocalyx, for example, increased endothelial and retinal syndecan-1, but decreased endothelial and retinal syndecan-3. This indicates that the reported decrease in the retinal glycocalyx in diabetes in not a result of a non-specific shedding mechanism. Moreover, mRNA measurements indicated a similar diversity, with increases in endothelial and/or retinal levels of syndecan-1, glypican-1, and CD44, but a decrease for syndecan-3, with these increases in mRNA potentially a compensatory reaction to the overall loss of glycocalyx.
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影响因子：
--
作者：
De Rossi G;Whiteford JR
通讯作者：
Whiteford JR
DOI：
10.1039/c3ib40199e
发表时间：
2014-03
期刊：
Integrative biology : quantitative biosciences from nano to macro
影响因子：
--
作者：
Ebong EE;Lopez-Quintero SV;Rizzo V;Spray DC;Tarbell JM
通讯作者：
Tarbell JM
影响因子：
3.4
作者：
Bartosch, Anne Marie W.;Mathews, Rick;Tarbell, John M.
通讯作者：
Tarbell, John M.
影响因子：
5
作者：
Brown, J;Reading, SJ;Brown, CA
通讯作者：
Brown, CA
DOI：
10.1152/ajpheart.00090.2018
发表时间：
2019-03-01
影响因子：
4.8
作者：
Ali, Mohamed M.;Mahmoud, Abeer M.;Phillips, Shane A.
通讯作者：
Phillips, Shane A.