硫化氢缓释供体ADT-OH对神经前体细胞增殖的调控  

Regulatory effects of ADT-OH on the proliferation of neural precursor cells

作　　者：周立宇[1] 马艳霞[2] 齐士斌 赛吉拉夫 韦善文 倪莉[1] Zhou Liyu;Ma Yanxia;Qi Shibin;Saijilafu;Wei Shanwen;Ni Li(Department of Orthopedics,The First Affiliated Hospital of Soochow University,Suzhou 215006,Jiangsu Province,China;Orthopedic Institute,Soochow University,Suzhou 215006,Jiangsu Province,China)

机构地区：[1]苏州大学附属第一医院骨科,江苏省苏州市215006 [2]苏州大学骨科研究所,江苏省苏州市215006

出　　处：《中国组织工程研究》2021年第1期96-100,共5页Chinese Journal of Tissue Engineering Research

基　　金：国家自然科学基金项目(81571189),项目负责人:赛吉拉夫。

摘　　要：背景:研究人员认为,硫化氢作为一种重要的细胞保护分子,通过人为调控内源性硫化氢生物合成或使用硫化氢供体体外给药可能成为新的疾病治疗方式,用来恢复病变细胞或器官系统的生理功能。ADT-OH是硫化氢的一种缓释供体,它能够提高谷氨酸诱导的损伤海马神经细胞的生存率,但是对大脑皮质神经前体细胞增殖的影响尚不清楚。目的:探究ADT-OH对胚胎期大脑皮质神经前体细胞增殖的影响。方法:分离E14.5 d胚胎小鼠大脑皮质心室带和脑室管膜下区神经前体细胞,将1只胎鼠的神经前体细胞接种于1个孔中(24孔板),培养基中加入100μmol/L ADT-OH药物进行培养,3 d后统计每孔中神经球的大小和数目,对培养的神经前体细胞进行BrdU标记检测细胞增殖率,用增殖细胞的特异性抗体Ki67对细胞进行免疫荧光染色进一步验证细胞增殖情况,最后采用Western blot检测增殖相关基因cyclin D1的表达。结果与结论:ADT-OH可促进神经球的形成,提高神经前体细胞的增殖率,同时神经前体细胞中增殖相关基因cyclin D1的表达上调。由以上数据可知,ADT-OH促进神经前体细胞的增殖可能是通过调控cyclin D1的表达来完成。BACKGROUND:Researchers believe that hydrogen sulfide(H2S),as an important cell protective molecule,may become a new treatment method to restore the physiological function of diseased cells or organ systems through the artificial regulation of endogenous H2S biosynthesis or in vitro administration of H2S donor.ADT-OH is a slow-release donor of H2S that can improve the survival rate of hippocampal nerve cells with glutamate-induced injury,but studies on the proliferation of cerebral cortical neural precursor cells are rare.OBJECTIVE:To investigate the effect of ADT-OH on the proliferation of neural precursor cells in embryonic cerebral cortex.METHODS:Neural precursor cells from cerebral cortical ventricular zone and subventricular zone of embryonic mice at embryonic 14.5 days were isolated.Neural precursor cells from one fetal mouse were inoculated into one well(24-well plate),and cultured with the medium containing 100μmol/L ADT-OH.The size and number of neural spheres per well were measured at 3 days after culture.The proliferation rate of cultured neural precursor cells was detected by BrdU labeling.The proliferation of the cells was further verified by immunofluorescence staining with the specific antibody Ki67.The expression of cyclin D1 was finally detected by western blot assay.RESULTS AND CONCLUSION:Our experimental results showed that ADT-OH could promote the formation of neural spheres,and further detection by BrdU and Ki67 antibody showed that ADT-OH could promote the proliferation rate of neural precursor cells.Meanwhile,the expression of cyclin D1,a proliferationrelated gene,was up-regulated in neural precursor cells after ADT-OH treatment.Overall,ADT-OH may promote the proliferation of neural precursor cells by regulating the expression of cyclin D1.

关 键 词：硫化氢 ADT-OH 神经前体细胞 细胞增殖 神经球 BRDU Ki67 免疫荧光染色 cyclin D1 

分 类 号：R459.9[医药卫生—治疗学] R394.2[医药卫生—临床医学]