Bhattacharya Sanjoy K, Gabelt B'Ann T, Ruiz Jose, Picciani Renata, Kaufman Paul L
Bascom Palmer Eye Institute, University of Miami, Miami, Florida 33136, USA.
Invest Ophthalmol Vis Sci. 2009 Feb;50(2):551-9. doi: 10.1167/iovs.08-2632. Epub 2008 Oct 3.
To determine the effect of transforming growth factor (TGF)-beta2 treatment on intraocular pressure (IOP), outflow facility, and cochlin expression in vitro in monkey and pig organ-cultured anterior segments (MOCAS and POCAS).
MOCAS (rhesus and cynomolgus) or POCAS were infused with media containing 10 ng/mL TGFbeta2 to one segment of each pair and 0.1% BSA (vehicle) to the contralateral segment for up to 14 days at a constant rate. Cochlin expression was determined by immunohistochemical study, ELISA, and Western blot analysis using chicken polyclonal antibodies against different regions of cochlin.
TGFbeta2 infusion produced elevated IOP in MOCAS (usually after 5 days), that was approximately 45% greater than baseline and compared to control segments. Outflow facility (OF) was decreased by approximately 40% compared with pretreatment baseline (n=5). In POCAS (n=7), IOP was increased (approximately 3 days) by approximately 75% compared with baseline and contralateral changes. The IOP elevation subsided thereafter. Cochlin levels increased with duration of TGFbeta2 treatment in the media and in the region of the trabecular meshwork in both species.
TGFbeta2-induced IOP elevation was associated with an increase in cochlin secretion into the media and expression in the tissue of MOCAS and POCAS. Whether cochlin overexpression contributes to elevated IOP or is a consequence of other changes relevant to IOP elevation remains to be determined.
确定转化生长因子(TGF)-β2处理对猴和猪器官培养眼前节(猴眼前节器官培养物和猪眼前节器官培养物)的眼压(IOP)、房水流出易度和耳蜗素表达的体外影响。
将含有10 ng/mL TGF-β2的培养基注入每对猴眼前节器官培养物(恒河猴和食蟹猴)或猪眼前节器官培养物的一个节段,将0.1%牛血清白蛋白(载体)注入对侧节段,以恒定速率持续14天。使用针对耳蜗素不同区域的鸡多克隆抗体,通过免疫组织化学研究、酶联免疫吸附测定(ELISA)和蛋白质印迹分析来确定耳蜗素表达。
注入TGF-β2使猴眼前节器官培养物的眼压升高(通常在5天后),比基线水平高约45%,并与对照节段相比。与预处理基线相比,房水流出易度降低了约40%(n = 5)。在猪眼前节器官培养物中(n = 7),与基线和对侧变化相比,眼压升高了约75%(约3天)。此后眼压升高消退。在这两个物种中,耳蜗素水平随着培养基中TGF-β2处理时间的延长以及小梁网区域的时间延长而增加。
TGF-β2诱导的眼压升高与耳蜗素分泌到培养基中增加以及猴眼前节器官培养物和猪眼前节器官培养物组织中的表达增加有关。耳蜗素过表达是否导致眼压升高或是否是与眼压升高相关的其他变化的结果仍有待确定。