Kubota Yasuhiro, Noguchi Hirofumi, Seita Masayuki, Yuasa Takeshi, Sasamoto Hiromi, Nakaji Shuhei, Okitsu Teru, Fujiwara Toshiyoshi, Kobayashi Naoya
Department of Gastroenterological Surgery, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences , Okayama , Japan.
† Department of Surgery, Chiba-East National Hospital, National Hospital Organization , Chiba , Japan.
Cell Med. 2013 Oct 21;6(1-2):15-23. doi: 10.3727/215517913X674199. eCollection 2013 Dec 30.
The number of patients with diabetes is on an increasing trend, thus leading to the belief that diabetes will be the largest medical problem of the 21st century. Islet transplantation can improve glycometabolic control in patients with type 1 diabetes. We studied the viability of Rho-associated protein kinase (ROCK) inhibitor Y-27632 in a culture system in vitro on freshly isolated rat islets. Islet isolation was conducted on a Lewis rat, and studies of culture solutions were split into two groups, one group using ROCK inhibitor Y-27632, and another without. On the seventh day of culture, we evaluated the differences for the cell morphology, viability, and insulin secretion. The Y-27632 group maintained form better than the group without Y-27632. With strong expression of Bcl-2 observed with the Y-27632 group, and expression suppressed with Bax, inhibition of apoptosis by Y-27632 was confirmed. The Y-27632 group predominantly secreted insulin. For islet transplantation, Y-27632 inhibited cell apoptosis in a graft and was also effective in promoting insulin secretion. We were able to confirm effective morphological and functional culture maintenance by separating islets from a rat and adding ROCK inhibitor Y-27632 to the medium.
糖尿病患者的数量呈上升趋势,因此人们认为糖尿病将成为21世纪最大的医学问题。胰岛移植可改善1型糖尿病患者的糖代谢控制。我们在体外培养系统中研究了Rho相关蛋白激酶(ROCK)抑制剂Y-27632对新鲜分离的大鼠胰岛的作用。在Lewis大鼠上进行胰岛分离,培养液研究分为两组,一组使用ROCK抑制剂Y-27632,另一组不使用。在培养的第7天,我们评估了细胞形态、活力和胰岛素分泌的差异。Y-27632组的形态维持优于无Y-27632组。Y-27632组观察到Bcl-2强表达,而Bax表达受到抑制,证实Y-27632抑制细胞凋亡。Y-27632组主要分泌胰岛素。对于胰岛移植,Y-27632抑制移植物中的细胞凋亡,并且在促进胰岛素分泌方面也有效。通过从大鼠分离胰岛并向培养基中添加ROCK抑制剂Y-27632,我们能够证实有效的形态和功能培养维持。