摘要
目的:探讨新生血管特异性结合肽GX1二聚体对视网膜新生血管生成的影响。方法:化学合成GX1二聚体、GX1单体、对照肽二聚体,通过CCK-8实验、管状结构形成实验、迁移实验研究GX1二聚体对大鼠视网膜内皮细胞(RMEC)增殖、微管形成、迁移能力的影响,流式细胞学技术分析其对细胞周期分布和凋亡的影响。结果:CCK-8结果显示,与对照肽二聚体及阴性对照组相比,100-200μM GX1二聚体及单体均可抑制RMEC增殖(P<0.05),且随着GX1二聚体及单体浓度升高,抑制作用逐渐增强,呈剂量依赖性;各浓度GX1二聚体均较单体抑制作用增强,并有统计学差异(P<0.05)。管状结构形成实验、细胞损伤迁移实验结果显示与对照肽二聚体及PBS组相比,GX1二聚体及GX1单体均可明显抑制视网膜内皮细胞管状结构的形成及迁移,且二聚体抑制作用强于单体;对照肽二聚体仅有轻微的抑制视网膜内皮细胞管状结构形成的作用,对细胞迁移无明显抑制作用。流式细胞术分析显示与对照肽及阴性对照组相比,GX1二聚体及GX1单体均可诱导细胞凋亡(P<0.05),且GX1二聚体的诱导作用强于GX1单体(P<0.05),而对细胞周期分布则无明显影响。结论:GX1二聚体和GX1单体均可抑制视网膜新生血管内皮细胞增殖、微管形成、迁移能力及诱导凋亡,且GX1二聚体较GX1单体作用增强。GX1二聚体有望代替单体成为糖尿病视网膜病变新生血管靶向治疗小肽类药物。
Objective: To evaluate the antiangiogenesis ability of dimeric GX1 to retinal angiogenesis. Methods: Dimeric GX1,GX1 monomer and dimeric control peptides were synthesized. Antiangiogenesis of dimeric GX1 on rat retinal microvasculature endothelial cells(RMEC) was analyzed by CCK-8 assay, migration assay, and tube formation assay. Flow cytometry assay was performed to evaluate the effects of dimeric GX1 to RMEC cycle and apoptosis. Results: CCK-8 assay showed that both dimeric GX1 and GX1 monomer compared to dimeric control peptides and PBS group had significant inhibitory effect on the proliferation of RMEC(P<0.05) from 100 to 200 μM in dose-dependent manner. Dimeric GX1 showed more significant inhibitory effect than GX1 monomer(P<0.05). Tube formation assay and cell migration assay showed that both dimeric GX1 and GX1 monomer could inhibit tube formation assay and cell migration of RMEC compared to dimeric control peptides and PBS group. Dimeric control peptides only showed slightly inhibitory effect on tube formation of RMEC as well as no effect on cell migration. Flow cytometry assay also showed that both dimeric GX1 and GX1 monomer could induce RMEC apoptosis compared to dimeric control peptides and PBS group(P<0.05), but dimeric GX1 had stronger induction effect than GX1 monomer(P<0.05). In addition, there is no significant effect on cell cycle distribution was observed. Conclusions: Dimeric GX1 owned more significant antiangiogenesis ability on RMEC than GX1 monomer. Dimeric GX1 was more promising to be explored for effective antiangiogenesis targeting drug to diabetic retinopathy.
作者
罗莹莹
刘惊涛
张海萍
雷志杰
崔巍
吴开春
惠晓丽
LUO Ying-ying;LIU Jing-tao;ZHANG Hai-ping;LEI Zhi-jie;CUI Wei;WU Kai-chun;HUI Xiao-li(Department of Geriatric endocrinology,First Affiliated Hospital of Xi'an Jiaotong University,Xi'an,Shaanxi,710061,China;Affiliated No.986 Hospital of Xijing Hospital,Air Force Military Medical University,Xi'an,Shaanxi,710054,China;Xijing Hospital of Digestive Diseases&State Key Laboratory of Cancer Biology,Air Force Military Medical University,Xi'an,Shaanxi,710032,China)
出处
《现代生物医学进展》
CAS
2020年第1期35-40,共6页
Progress in Modern Biomedicine
基金
国家自然科学创新研究群体项目(81421003)
国家自然科学基金项目(30900704)
陕西省重点研发计划一般项目(2017SF-104,2018SF-161)
陕西省科技攻关项目(2013K12-09-03)
陕西省卫生计生科研基金项目(2016D066)
西安市科技计划项目(201805095YX3SF296)。
