-
糖尿病视网膜病变是糖尿病最常见的微血管并发症之一,是发达国家和部分发展中国家工作人群致盲的重要原因。2000年,全球糖尿病病人达1.71亿,到2030年,全球糖尿病病人预计达到3.6亿。目前,中国糖尿病病人已达9 240万,居世界之首,而糖尿病视网膜病变患病率达37%[1]。因而,糖尿病视网膜病变机制的研究及其防治是全球眼科学和内分泌专业的学者共同关注的问题。
视网膜屏障功能受损,是糖尿病视网膜病变发生发展的重要病理基础。视网膜色素上皮(retinal pigment epithelium,RPE)细胞是视网膜外屏障的重要组成细胞,其通过细胞间及细胞与基质间的黏附构成的紧密连接在维持视网膜的内环境中发挥重要作用。由于RPE细胞的特殊位置,极易受到高血糖的影响[2-3]。syndecan-1是一种黏附分子,属于整联蛋白跨膜硫酸肝素蛋白多糖家族成员,可调控细胞的黏附、移行和增生,参与组织器官分化发育、血管形成、组织再生、组织损伤修复、炎症、脂质代谢等一系列病理生理过程[4-6]。syndecan-1分子可调控RPE细胞的黏附、增生和移行[7],并参与糖尿病视网膜病变的发生发展[8-9]。目前,国内外鲜见高糖对于RPE细胞syndecan-1表达影响的研究报道。本实验的目的是观察高糖对体外培养的人RPE细胞syndecan-1表达的影响。现作报道。
高糖对人视网膜色素上皮细胞syndecan-1表达的影响
Effect of high glucose on the expression level of syndecan-1 in human retinal pigment epithelial cells
-
摘要:
目的检测高糖对视网膜色素上皮(RPE)细胞syndecan-1表达的影响。 方法25 mmol/L高糖刺激人RPE细胞4 h、12 h和24 h,分别采用免疫组织化学和蛋白印迹法检测细胞syndecan-1蛋白的表达,采用实时定量聚合酶链反应检测syndecan-1 mRNA的表达。 结果高糖刺激后,RPE细胞排列紊乱,细胞体变长变大,脱落细胞多见。正常RPE细胞表达syndecan-1,且定位于细胞质。高糖刺激4 h,RPE细胞syndecan-1的蛋白和mRNA表达开始下降,12 h时表达降至最低,24 h时syndecan-1的表达基本回复至正常水平(P < 0.01)。 结论高糖可短暂下调RPE细胞syndecan-1蛋白和mRNA的表达。 -
关键词:
- 视网膜色素上皮细胞 /
- 高血糖 /
- syndecan-1
Abstract:ObjectiveTo detect the effects of high glucose on the expression level of syndecan-1 in human retinal pigment epithelial(RPE) cells. MethodsThe RPE cells were treated with 25 mmol/L glucose for 4 h, 12 h and 24 h, respectively.The protein and mRNA expression levels of syndecan-1 were detected using immunohistochemistry staining and Western blot, and quantitative real-time polymerase chain reaction(qPCR), respectively. ResultsAfter exposure to high glucose, the RPE cells were arranged in disorder, the cell bodies became longer and larger, the shedding cells were easily seen.The syndecan-1 expression was detected in normal RPE cells, and located in cytoplasm.After 4 h of treatment with high glucose, the expression levels of syndecan-1 protein and mRNA decreased, decreased to the minimum at 12 h, and basically returned to the normal level at 24 h(P < 0.01). ConclusionsHigh glucose can transiently down-regulate the syndecan-1 protein and mRNA expression in RPE cells. -
Key words:
- retinal pigment epithelial cell /
- high glucose /
- syndecan-1
-
[1] 王素常, 李甦雁.增生型糖尿病视网膜病变病人泪液分泌的研究[J].徐州医学院学报, 2017, 37(5):315. doi: 10.3969/j.issn.1000-2065.2017.05.008 [2] 韩小霞, 惠延年, 宋虎平, 等.曲安奈德对高糖培养人视网膜色素上皮细胞细胞间黏附分子-1表达的影响[J].国际眼科杂志, 2006, 6(3):596. doi: 10.3969/j.issn.1672-5123.2006.03.025 [3] 姚毅, 关明, 赵秀琴, 等.缺氧和高浓度葡萄糖对体外培养人视网膜色素上皮衍生因子表达的影响[J].中华医学杂志, 2003, 83(22):1989. doi: 10.3760/j:issn:0376-2491.2003.22.015 [4] WANG JB, ZHANG YJ, GUAN J, et al.Enhanced syndecan-1 expression on neutrophils in patients with type 2 diabetes mellitus[J].Acta Diabetol, 2012, 49(1):41. doi: 10.1007/s00592-011-0265-1 [5] WANG JB, ZHANG YJ, ZHANG Y, et al.Negative correlation between serum syndecan-1 and apolipoprotein A1 in patients with type 2 diabetes mellitus[J].Acta Diabetol, 2013, 50(2):111. doi: 10.1007/s00592-010-0216-2 [6] WANG JB, GUAN J, SHEN J, et al.Insulin increases shedding of syndecan-1 in the serum of patients with type 2 diabetes mellitus[J].Diabetes Res Clin Pract, 2009, 86(2):83. doi: 10.1016/j.diabres.2009.08.002 [7] 王静波, 惠延年, 韩泉洪, 等.细胞因子对人视网膜色素上皮细胞syndecan-1的影响[J].中华眼底病杂志, 2006, 22(2):113. doi: 10.3760/j.issn:1005-1015.2006.02.013 [8] 王静波, 宋宗明, 孙吉君.糖尿病视网膜病变病人玻璃体液中syndecan-1及上皮中性粒细胞活化肽78的表达[J].临床眼科杂志, 2013, 21(6):481. [9] 王静波, 余欢龙.syndecan-1在糖尿病视网膜中的表达[J].国际眼科杂, 2013, 13(4):660. [10] 王雨生, 严密, 扬扶华.视网膜色素上皮细胞培养技术及其应用[J].中华眼底病杂志, 1994, 10(2):124. [11] 韩小霞, 郭长梅, 惠延年, 等.曲安奈德对高糖培养RPE细胞ICAM-1和ILK表达的影响[J].眼科新进展, 2010, 30(12):1111. [12] 毕文娇, 李睿姝, 侯定善, 等.高糖对人视网膜色素上皮细胞神经钙黏连蛋白和纤维连接蛋白表达的影响[J].国际眼科杂志, 2014, 14(9):1578. [13] 许艺民, 冯云, 马志中.PDR、PVR增生膜组织超微结构及生长因子受体表达生物学特点的比较研究[J].眼科研究, 2005, 23(2):147. doi: 10.3760/cma.j.issn.2095-0160.2005.02.010 [14] 蔡莉, 易敬林.视网膜色素上皮细胞损伤机制研究进展[J].眼科新进展, 2012, 32(9):898. [15] 王静波, 惠延年, 韩泉洪, 等.细胞因子对人视网膜色素上皮细胞syndecan-1表达的影响[J].中华眼底病杂志, 2006, 22(2):113. doi: 10.3760/j.issn:1005-1015.2006.02.013 [16] 王静波, 惠延年, 马吉献, 等.syndecan-1在增生性玻璃体视网膜病变增生膜中的表达[J].眼科研究, 2004, 22:474. doi: 10.3760/cma.j.issn.2095-0160.2004.05.035 [17] WANG JB, TIAN CW, GUO CM, et al.Increased levels of soluble syndecan-1 in the subretinal fluid and the vitreous of eyes with rhegmatogenous retinal detachment[J].Curr Eye Res, 2008, 33(1):101. doi: 10.1080/02713680701809272 [18] 胡永亮, 王静波, 周历, 等.胰岛素对人视网膜微血管内皮细胞syndecan-1的表达及细胞通透性和增殖的影响[J].国际眼科杂志, 2018, 18(8):1381. [19] ALEXOPOULOU AN, MULTHAUPT HA, COUCHMAN JR.Syndecans in wound healing, inflammation and vascular biology[J].Int J Biochem Cell Biol, 2007, 39(3):505. doi: 10.1016/j.biocel.2006.10.014 [20] KATO M, SAUNDERS S, NGUYEN H, et al.Loss of cell surface syndecan-1 causes epithelial cells to transform into anchorage-independent mesenchyme-like cells[J].Mol Biol Cell, 1995, 6:559. doi: 10.1091/mbc.6.5.559