Departments of Gastrointestinal Surgery, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, 1277 Jiefang Avenue, Wuhan, Hubei, 430022, People's Republic of China,
Mol Neurobiol. 2013 Dec;48(3):429-37. doi: 10.1007/s12035-013-8430-x. Epub 2013 Mar 20.
To explore a potential methodology for treating aganglionic megacolon, neural stem cells (NSCs) expressing engineered endothelin receptor type B (EDNRB) and glial cell-derived neurotrophic factor (GDNF) genes were transplanted into the aganglionic megacolon mice. After transplantation, the regeneration of neurons in the colon tissue was observed, and expression levels of differentiation-related genes were determined. Primary culture of NSCs was obtained from the cortex of postnatal mouse brain and infected with recombinant adenovirus expressing EDNRB and GDNF genes. The mouse model of aganglionic megacolon was developed by treating the colon tissue with 0.5 % benzalkonium chloride (BAC) to selectively remove the myenteric nerve plexus that resembles the pathological changes in the human congenital megacolon. The NSCs stably expressing the EDNRB and GDNF genes were transplanted into the benzalkonium chloride-induced mouse aganglionic colon. Survival and differentiation of the implanted stem cells were assessed after transplantation. Results showed that the EDNRB and GDNF genes were able to be expressed in primary culture of NSCs by adenovirus infection. One week after implantation, grafted NSCs survived and differentiated into neurons. Compared to the controls, elevated expression of EDNRB and GDNF was determined in BAC-induced aganglionic megacolon mice with partially improved intestinal function. Those founding indicated that the genes transfected into NSCs were expressed in vivo after transplantation. Also, this study provided favorable support for the therapeutic potential of multiple gene-modified NSC transplantation to treat Hirschsprung's disease, a congenital disorder of the colon in which ganglion cells are absent.
为了探索一种治疗先天性巨结肠症的潜在方法，将表达工程化内皮素受体 B 型（EDNRB）和胶质细胞源性神经营养因子（GDNF）基因的神经干细胞（NSCs）移植到先天性巨结肠症小鼠的无神经节细胞巨结肠中。移植后，观察结肠组织中神经元的再生情况，并测定分化相关基因的表达水平。从新生鼠大脑皮质中获得原代培养的 NSCs，并感染表达 EDNRB 和 GDNF 基因的重组腺病毒。通过用 0.5%苯扎氯铵（BAC）处理结肠组织来制备先天性巨结肠症小鼠模型，以选择性地去除类似于人类先天性巨结肠病变的肌间神经丛。将稳定表达 EDNRB 和 GDNF 基因的 NSCs 移植到 BAC 诱导的先天性巨结肠症小鼠的无神经节细胞巨结肠中。移植后评估植入干细胞的存活和分化情况。结果表明，腺病毒感染可使 EDNRB 和 GDNF 基因在原代 NSCs 中表达。移植后 1 周，移植的 NSCs 存活并分化为神经元。与对照组相比，在 BAC 诱导的先天性巨结肠症小鼠中，EDNRB 和 GDNF 的表达水平升高，部分改善了肠道功能。这些发现表明，移植后 NSCs 中转染的基因在体内得到表达。此外，该研究为多基因修饰 NSC 移植治疗先天性巨结肠症的治疗潜力提供了有利支持，先天性巨结肠症是一种结肠先天性疾病，其中缺乏神经节细胞。
