改良透明质酸水凝胶控释脑源性神经营养因子对大鼠胚胎神经干细胞生长、分化和凋亡的影响  

Effect of modified hyaluronic acid hydrogel controlled release of brain derived neurotrophic factor on growth,differentiation and apoptosis of embryonic neural stem cells in rats

作　　者：王震 张香路 李阳 阮铭暄 黄斐[1] Wang Zhen;Zhang Xianglu;Li Yang;Ruan Mingxuan;Huang Fei(Dept of Orthopedics, The Fourth Affiliated Hospital of Anhui Medical University,Hefei 230012)

机构地区：[1]安徽医科大学第四附属医院骨科,合肥230012

出　　处：《安徽医科大学学报》2022年第5期805-811,共7页Acta Universitatis Medicinalis Anhui

基　　金：国家自然科学基金(编号:81601974);安徽省转化医学研究院科研基金(编号:2021zhyx-c53)。

摘　　要：目的研究改良透明质酸(HA)水凝胶控释脑源性神经营养因子(BDNF)对大鼠胚胎神经干细胞(rFNSCs)生长、分化及凋亡的影响。方法实验以大鼠胚胎干细胞为研究对象,将实验分为3组,分别为空白对照组[rFNSCs与聚乳酸乙醇酸(PLGA)和HA共培养]、条件对照组(rFNSCs与BDNF-PLGA和HA共培养)、实验组(rFNSCs与BDNF-PLGA和改良HA水凝胶即Dex-HA共培养)。采用ELISA法检测控释支架中BDNF的释放曲线;Live/Dead实验检测各组中的rFNSCs凋亡情况;CCK-8法检测rFNSCs与水凝胶共培养后的细胞增殖活性;免疫细胞化学染色法检测不同组中rFNSCs的分化情况。结果ELISA法结果显示水凝胶对BDNF的控释可以持续14 d。Live/Dead实验显示实验组中rFNSCs凋亡数低于空白对照组和条件对照组。CCK-8实验表明,三组中实验组rFNSCs细胞增殖活性最强,差异有统计学意义(P<0.05)。免疫细胞化学染色结果表明,与空白对照组和条件对照组相比,实验组水凝胶在体外能促进rFNSCs向神经元分化,减少向星形胶质细胞的分化,差异有统计学意义(P<0.05)。结论改良HA水凝胶控释BDNF可以促进rFNSCs的生长、分化并减少其凋亡。Objective To investigate the effect of modified hyaluronic acid(HA)hydrogel-controlled release brain-derived neurotrophic factor(BDNF)on the growth,differentiation,and apoptosis of rat embryonic neural stem cells(rFNSCs).Methods The experiment took rat embryonic stem cells as the research object.The experiments were divided into three groups,blank control group[rFNSCs co-cultured with poly(lactic-co-glycolic acid)(PLGA)and HA],conditional control group(rFNSCs co-cultured with BDNF-PLGA and HA),and experimental group(rFNSCs co-cultured with BDNF-PLGA and modified hyaluronic acid hydrogel that is Dex-HA).ELISA was used for detecting the release curve of BDNF in the controlled-release scaffold.The Live/Dead assay detected the apoptosis of rFNSCs in each group,CCK-8 experiment was used to test the cell proliferation activity of rFNSCs after coculture with hydrogel.The differentiation ratios of rFNSCs in different groups were detected by immunocytochemical staining.Results ELISA result showed that the controlled-release scaffold could release BDNF for 14 days.Live/Dead result showed the number of rFNSCs apoptotic in experimental group was less than blank control group and conditional control group.CCK-8 experiment result showed that the cell proliferation activity of rFNSCs in the experimental group was showed stronger survivability than blank control group and conditional control group.The difference was statistically significant(P<0.05).Moreover,immunocytochemical staining showed that the experimental group hydrogel could promote the neuronal differentiation of rFNSCs and reduce their differentiation into astrocytes in vitro compared with the blank control group and the conditional control group.The difference was statistically significant(P<0.05).Conclusion modified hyaluronic acid hydrogel controlled release brain-derived neurotrophic factor can promote the growth and differentiation of rFNSCs and reduce their apoptosis.

关 键 词：透明质酸水凝胶 胚胎神经干细胞 脑源性神经营养因子 

分 类 号：R394.2[医药卫生—医学遗传学]