Dynamic Alteration of Low-Density Lipoprotein Receptor After Exposure to Transforming Growth Factor-β2 in Human Tenon's Capsule Fibroblasts
Dynamic Alteration of Low-Density Lipoprotein Receptor After Exposure to Transforming Growth Factor-β2 in Human Tenon's Capsule Fibroblasts
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DOI：
10.1089/jop.2009.0042
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发表时间：
2009-12-01
影响因子：
2.3
通讯作者：
Ge, Jian
中科院分区：
文献类型：
--
作者：
Shao, Tingting;Gao, Qianying;Ge, Jian
目的：本研究探讨人Tenon's囊成纤维细胞暴露于转化生长因子-β（2）（TGF-β（2））后低密度脂蛋白受体的动态变化及其对低密度脂蛋白（LDL）的结合和摄取。方法：培养取自择期白内障手术患者的Tenon's囊成纤维细胞，并用不同浓度（0.1-10ng/mL）的TGF-β（2）刺激。 24、48 和 72 小时。分别通过相对定量实时RT-PCR和蛋白质印迹分析来分析LDLr mRNA和蛋白质水平。通过共聚焦显微镜评估 DiO (3,3'-二十八烷基氧羰花青) 标记的 LDL 的结合和摄取。 结果：实时 RT-PCR 和 Western blot 分析显示相似的结果，表明暴露于 TGF-β(2) 后，LDLr 的蛋白和 mRNA 的表达以浓度依赖性和时间依赖性的方式发生，在 Tenon 囊中 72 h 时在浓度为 1.0 ng/mL 时出现峰值成纤维细胞。共聚焦显微镜显示DiO-LDL结合和摄取具有时间依赖性，大约在6小时时达到饱和。结论：本研究表明，暴露于TGF-β(2)后，LDLrs在活化的Tenon's囊成纤维细胞中以浓度依赖性和时间依赖性的方式过度表达。结果表明，活化的Tenon's囊成纤维细胞中的LDLr可能成为控制药物递送的靶受体的新焦点，特别是在结膜伤口过度愈合过程中的抗疤痕治疗中。
Purpose: The present study investigated dynamic alteration of low-density lipoprotein receptor and its binding and uptake of low-density lipoprotein (LDL) after exposure to transforming growth factor-beta(2) (TGF-beta(2)) in human Tenon's capsule fibroblasts.Methods: Tenon's capsule fibroblasts obtained from elective cataract surgery patients were cultured and stimulated with different concentrations (0.1-10 ng/mL) of TGF-beta(2) for 24, 48, and 72 h. The LDLr mRNA and protein levels were analyzed by relative quantification real-time RT-PCR and Western blot analysis, respectively. The binding and uptake of DiO (3,3'-dioctadecyloxacarbocyanine)-labeled LDL was assessed by confocal microscopy.Results: Real-time RT-PCR and Western blot analyses showed similar results revealing that after exposure to TGF-beta(2), the expression of protein and mRNA of LDLr occurred in a concentration-dependent and time-dependent manner with a peak at a concentration of 1.0 ng/mL at 72 h in Tenon's capsule fibroblasts. Confocal microscopy showed that DiO-LDL binding and uptake were time-dependent, reaching saturation at similar to 6 h.Conclusions: This study shows that LDLrs were overexpressed in the activated Tenon's capsule fibroblasts in a concentration-dependent and time-dependent manner after exposure to TGF-beta(2). The results suggest that LDLr in the activated Tenon's capsule fibroblasts may become a novel focus as a target receptor for controlled drug delivery, particularly in anti-scarring therapy during excessive conjunctival wound healing.