Fuchshofer Rudolf, Kottler Ulrike B, Ohlmann Anne V, Schlötzer-Schrehardt Ursula, Jünemann Anselm, Kruse Friedrich E, Ohlmann Andreas
Institute of Human Anatomy and Embryology, University of Regensburg, Regensburg, Germany.
Mol Vis. 2011 Jan 19;17:177-85.
To investigate the expression of the matricellular protein SPARC (secreted acidic cysteine-rich glycoprotein) in scarred human Tenon's capsule and in cultured human Tenon's fibroblasts (HTF), and to analyze the influence of SPARC on cell proliferation and collagen matrix contraction in vitro.
Human Tenon's capsule scars obtained from surgical revisions after filtration surgery were analyzed for SPARC expression by immunohistochemistry. In cultured HTF cells, SPARC expression was assessed by northern and western blot analyses after incubation with transforming growth factor (TGF)-β1 and TGF-β2. Cell proliferation was determined by bromodeoxyuridine (BrdU)-labeling and HTF cells-mediated collagen matrix contraction by morphometric measurements of three-dimensional collagen lattices after treatment with SPARC and/or TGF-β1.
In scarred human Tenon's capsule specimens, an increased expression of SPARC was mainly localized to the extracellular matrix and to blood vessel walls as compared to healthy control Tenon's capsule. In cultured HTF cells, treatment with TGF-β1 more than TGF-β2 induced the expression of SPARC both on the mRNA and protein level. Incubation of HTF cells with SPARC resulted in an increase in collagen matrix contraction and cell proliferation. Moreover, a combined incubation of SPARC and TGF-β1 stimulated HTF cell proliferation significantly over the levels that were observed after single treatment.
Our data provide evidence that SPARC contributes to excessive wound healing and scar formation in human Tenon's capsules after filtration surgery and may thus represent a novel target for anti-fibrotic strategies.
研究基质细胞蛋白SPARC（分泌性富含酸性半胱氨酸糖蛋白）在人瘢痕化的眼球筋膜囊及培养的人眼球筋膜成纤维细胞（HTF）中的表达，并分析SPARC对体外细胞增殖和胶原基质收缩的影响。
通过免疫组织化学分析滤过性手术后手术翻修获取的人眼球筋膜囊瘢痕中SPARC的表达。在培养的HTF细胞中，用转化生长因子（TGF）-β1和TGF-β2孵育后，通过Northern印迹和Western印迹分析评估SPARC的表达。用溴脱氧尿苷（BrdU）标记法测定细胞增殖，并用形态计量学方法测量经SPARC和/或TGF-β1处理后的三维胶原晶格，以检测HTF细胞介导的胶原基质收缩。
与健康对照眼球筋膜囊相比，在人瘢痕化眼球筋膜囊标本中，SPARC表达增加主要定位于细胞外基质和血管壁。在培养的HTF细胞中，TGF-β1比TGF-β2更能诱导SPARC在mRNA和蛋白质水平上表达。用SPARC孵育HTF细胞导致胶原基质收缩和细胞增殖增加。此外，SPARC和TGF-β1联合孵育显著刺激HTF细胞增殖，其水平高于单次处理后观察到的水平。
我们的数据表明，SPARC在滤过性手术后的人眼球筋膜囊中促成过度伤口愈合和瘢痕形成，因此可能是抗纤维化策略的一个新靶点。
