摘要
目的 探讨乳脂肪球表皮生长因子8 (MFG-E8)在青光眼大鼠视神经保护中的作用与机制。方法 构建青光眼大鼠模型,将MFG-E8或D89E注射至大鼠玻璃体腔内。检测大鼠的眼压变化。通过HE染色、TUNEL染色、免疫荧光染色分别检测大鼠视网膜组织病理损伤、节细胞凋亡和小胶质细胞激活水平;通过Western blotting检测视网膜组织中cleaved caspase-3、caspase-3、cleaved caspase-9、caspase-9和BAX蛋白表达;通过ELISA和实时定量PCR检测视网膜组织中IL-10、TGF-β和NGF的表达水平。结果 与对照组相比,青光眼大鼠的眼压显著增加,视网膜神经节细胞复合体(GCC)层变薄,凋亡节细胞增加,cleaved-caspase 3/caspase-3、cleaved caspase-9/caspase-9和BAX水平升高,IBA1阳性细胞数增加,且IL-10、TGF-β、NGF水平增高;经MFG-E8治疗后,青光眼大鼠视网膜GCC层厚度增加,cleaved caspase-3/caspase-3、cleaved caspase-9/caspase-9、BAX水平降低,IBA1阳性细胞数和IL-10、TGF-β、NGF水平均增加;而MFG-E8失活异构体D89E处理后,大鼠青光眼进展进一步加重。结论 MFG-E8能够介导小胶质细胞激活,抑制神经节细胞凋亡,减缓大鼠青光眼的进展。
Objective To investigate the role and mechanism of milk fat globule-EGF factor 8(MFG-E8)in neuroprotection in glaucoma rats.Methods A glaucoma rat model was constructed,and MFG-E8 or D89E was injected into the vitreous cavity of the rats.The intraocular pressure of the rats was measured.HE staining,TUNEL staining and immunofluorescence staining were used to detect the pathological injury,apoptosis and microglia activation of the retina.The protein expressions of cleaved caspase-3,caspase-3,cleaved caspase-9,caspase-9 and Bax were detected by Western blotting,and the expression levels of IL-10,TGF-βand NGF were detected by ELISA and RT-qPCR.Results Compared with the control group,the intraocular pressure of glaucoma rats significantly increased,the retinal GCC layer became thinner,the apoptotic cells increased,the levels of cleaved caspase-3/caspase-3,cleaved caspase-9/caspase-9 and Bax increased,and the number of IBA1 positive cells increased,after treatment with MFG-E8,retinal GCC layer thickness increased,and the levels of cleaved caspase-3/caspase-3,cleaved caspase-9/caspase-9,Bax decreased,the number of IBA1 positive cells and the levels of IL-10,TGF-β,NGF increased,but the progression of glaucoma was aggravated after the treatment of MFG-E8 inactivated isomer D89E.Conclusion MFG-E8 can mediate microglia activation,inhibit apoptosis of ganglion cells,and slow down the progression of glaucoma in rats.
作者
杨静
曾明兵
杨军
史贻玉
陈海波
YANG Jing;ZENG Mingbing;YANG Jun;SHI Yiyu;CHEN Haibo(Ward 1 of Hainan Eye Hospital and Key Laboratory of Ophthalmology,Zhongshan Ophthalmic Center,Sun Yat-sen University,Haikou 570311,China;Cataract Department,Hainan Aier Ehope Eye Hospital,Haikou 570226,China)
出处
《中国医科大学学报》
CAS
北大核心
2024年第7期591-596,共6页
Journal of China Medical University
基金
海南省自然科学基金青年基金(820QN414)
海南省卫生健康行业科研项目(20A200322)。
