McColm Janet R, Geisen Pete, Peterson Lynda J, Hartnett M Elizabeth
Department of Ophthalmology, University of North Carolina, 6135 Neuroscience Research Building, 103 Mason Farm Road, Chapel Hill, NC 27599-7041, USA.
Exp Eye Res. 2006 Aug;83(2):438-46. doi: 10.1016/j.exer.2006.01.027. Epub 2006 Apr 27.
To study the effect of leukemia inhibitory factor (LIF) on rat retinal vascular development, Sprague-Dawley rats at postnatal age 3 days (p3) were given intraperitoneal (IP) LIF and analysis performed at p6 (p3/6). p7 rats were given intravitreous (IV) LIF and analysis performed at p9 (p7/9). Control animals were PBS injected. At the time of analysis retinal flatmounts were prepared and stained with Griffonia lectin and activated caspase-3. The retinal peripheral avascular area was measured and number of apoptotic cells counted. In vitro, human retinal microvascular endothelial cells (RMVECs) were cultured in media containing LIF, with and without neutralizing antibody to LIF. Cells were stained with activated caspase-3 and apoptotic cells counted. Proliferation was measured by counting cell numbers, and cell cycle stage was determined using propidium iodide staining and FACS analysis. LIF injected either IP or IV had no effect on body weight or total retina area, but significantly increased the peripheral retinal avascular area. In both IP and IV injected groups there was no difference in the number of apoptotic cells between PBS- or LIF-injected groups; although in the p7/9 retinas, both injected groups had significantly more apoptotic cells than the non-injected group. In vitro, there was no effect of LIF on RMVEC apoptosis; however, cell counts were significantly lower in the LIF-treated group. Antibody to LIF restored the cell counts to untreated levels. LIF reduced the number of cells in S phase. LIF attenuates retinal vascular development in vivo through growth arrest, and not apoptosis, of endothelial cells.
为研究白血病抑制因子（LIF）对大鼠视网膜血管发育的影响，对出生3天（p3）的Sprague-Dawley大鼠进行腹腔内（IP）注射LIF，并在p6（p3/6）时进行分析。对p7大鼠进行玻璃体内（IV）注射LIF，并在p9（p7/9）时进行分析。对照组动物注射磷酸盐缓冲液（PBS）。在分析时，制备视网膜平铺片，并用 Griffonia 凝集素和活化的半胱天冬酶-3进行染色。测量视网膜周边无血管区，并计数凋亡细胞数量。在体外，将人视网膜微血管内皮细胞（RMVECs）培养在含有LIF的培养基中，分别添加和不添加LIF中和抗体。用活化的半胱天冬酶-3对细胞进行染色，并计数凋亡细胞。通过计数细胞数量测量增殖情况，并使用碘化丙啶染色和流式细胞术分析确定细胞周期阶段。腹腔内或玻璃体内注射LIF对体重或视网膜总面积没有影响，但显著增加了视网膜周边无血管区。在腹腔内和玻璃体内注射组中，注射PBS或LIF的组之间凋亡细胞数量没有差异；尽管在p7/9视网膜中，两个注射组的凋亡细胞均明显多于未注射组。在体外，LIF对RMVEC凋亡没有影响；然而，LIF处理组的细胞计数显著降低。LIF抗体将细胞计数恢复到未处理水平。LIF减少了处于S期的细胞数量。LIF在体内通过使内皮细胞生长停滞而非凋亡来减弱视网膜血管发育。
