高迁移率族B1(HMGB-1)蛋白在鼠视网膜新生血管形成中的调控作用

目的　探讨高迁移率族Ｂ１（ｈｉｇｈｍｏｂｉｌｉｔｙｇｒｏｕｐｂｏｘ-１，ＨＭＧＢ-１）蛋白在鼠视网膜新生血管形成中的调控作用。方法　将基质胶（Ｍａｔｒｉｇｅｌ）和ＨＭＧＢ-１混合物注入鼠龄为７ｄ（Ｐ７）的Ｃ５７ＢＬ／６Ｊ健康小鼠左眼视网膜下，基质胶与ＰＢＳ混合物注入右眼球作为对照组。注入１０ｄ后对获得的视网膜组织切片进行ＨＥ染色，计数突破视网膜前膜的血管内皮细胞核数。选用Ｃ５７ＢＬ／６Ｊ健康新生鼠建立氧诱导视网膜病变（ｏｘｙｇｅｎ-ｉｎｄｕｃｅｄｒｅｔｉｎｏｐａｔｈｙ，ＯＩＲ）动物模型，出生后第７天（Ｐ７）放入高氧环境中连续饲养５ｄ，然后置入正常空气中继续饲养；正常空气对照组小鼠则在空气中饲养。在Ｐ１７采用ＤｉＩ荧光造影视网膜铺片观察ＯＩＲ模型组和正常空气对照组小鼠视网膜血管形态变化；同时采用Ｗｅｓｔｅｒｎ-ｂｌｏｔ检测模型组和对照组小鼠玻璃体中ＨＭＧＢ-１蛋白的表达水平。结果　ＨＭＧＢ-１基质胶模型组与ＰＢＳ基质胶对照组基质胶区域突破视网膜内界膜的血管内皮细胞核数分别为（４２．１９±１．７６）个、（３１．２４±１．２５）个，经过ＨＭＧＢ-１与基质胶混合物处理的眼球中新生血管水平显著高于对照组（ｔ＝－４２．４２，Ｐ＝０. ０００）。ＤｉＩ荧光造影视网膜铺片结果显示，ＯＩＲ模型组鼠视网膜自视盘向中周部出现大片无灌注区，无灌注区周围出现新生血管丛；Ｗｅｓｔｅｒｎ-ｂｌｏｔ分析显示，与正常空气对照组相比，ＯＩＲ模型组鼠玻璃体中ＨＭＧＢ-１蛋白表达增加了７８％ ±７％，出现了显著上调（ｔ＝－４７．９０，Ｐ＝０．０００）。结论　ＨＭＧＢ-１在视网膜新生血管形成中起重要的调控作用。

Mediation effects of high mobility group B1 protein on retinal neovascularization in mice

Objective To investigate the mediation effects of high mobility group B1 ( HMGB-I) protein on retinal neovascularization in mice. Methods HMGB-I in phosphate buffered saline ( PBS) was nuxed with matrigel, and matrigel diluted with PBS only was used as a control. The matrigel with HMGB-I and PBS only was slowly injected to the subretinal of P7 C57BL/6J mice to form the matrigel induced model of neovascularization. And retinal neovascularization level of P17 matrigel induced model mice was quantified by counting the endothelial nuclei protruding into the vitreous cavity by HE stairung. C57BL/6J mice were used to form the oxygen-induced retinopathy ( OIR) model. Mice were exposed t0 75% oxygen tension from P7 to P12. And then they were returned to normal air and maintained for another 5 days ( P17 ) . Normoxia control mice of age identical with the hyperoxia group were maintained at room air. Retinal neovascularization was observed by fluorescence angiography at P17. Western blot analysis was also used to determine protein levels of HMGB-I in the retinal of mice at P17. Results The average endothelial cells which breaking through the inner limiting membrane in HMGBI with matrigel group and PBS with matrigel group were 42. 19 + 1. 76 ,31. 24 + 1. 25 ,respectively. The level of neovascularization in the retina of HMGB-I with matrigel group was higher than that of PBS with matrigel group ( t = - 42. 42 ,P = 0. 000) . The Dil fluorescence angiography of the mice showed that the non-perfusion area appeared from optic disc to peripheral portion in OIR model group , and a large scale of neovascular plexus appeared in this non-perfusion area. Western-blot analysis showed that the HMGB-I protein expression in OIR model group had a 78% +7% increase compared with control arumals ( t = - 47. 90 .P = 0. 000) . Conclrision HMGB-I plays an significant role in retinal neovascularization.