Pepper Andrew R, Bruni Antonio, Pawlick Rena, Wink John, Rafiei Yasmin, Gala-Lopez Boris, Bral Mariusz, Abualhassan Nasser, Kin Tatsuya, Shapiro A M James
1 Clinical Islet Transplant Program, Alberta Diabetes Institute, University of Alberta, Edmonton, AB, Canada. 2 Department of Surgery, University of Alberta, Edmonton, AB, Canada.
Transplantation. 2017 Oct;101(10):2321-2329. doi: 10.1097/TP.0000000000001638.
Islet transplantation is an effective therapy in type 1 diabetes and recalcitrant hypoglycemia. However, there is an ongoing need to circumvent islet loss posttransplant. We explore herein the potential of the pan-caspase inhibitor F573 to mitigate early apoptosis-mediated islet death within portal and extrahepatic portal sites in mice.
Mouse or human islets were cultured in standard media ±100 μM F573 and subsequently assessed for viability and apoptosis via terminal deoxynucleotidyl transferase dUTP nick end labeling staining and caspase-3 activation. Diabetic mice were transplanted with syngeneic islets placed under the kidney capsule (KC) or into the subcutaneous deviceless (DL) site at a marginal islet dose (150 islets), or into the portal vein (PV) at a full dose (500 islets). Human islets were transplanted under the KC of diabetic immunodeficient mice at a marginal dose (500 islet equivalents). Islets were cultured in the presence of F573, and F573 was administered subcutaneously on days 0 to 5 posttransplant. Control mice were transplanted with nontreated islets and were injected with saline. Graft function was measured by nonfasting blood glucose and glucose tolerance testing.
F573 markedly reduced human and mouse islet apoptosis after in vitro culture (P < 0.05 and P < 0.05, respectively). Furthermore, F573 improved human islet function when transplanted under the KC (P < 0.05); whereas F573 did not enhance murine islet marginal KC transplants. Conversely, F573 significantly improved mouse islet engraftment in the PV and DL site (P < 0.05 and P < 0.05, respectively).
The pan-caspase inhibitor F573 markedly reduces human and mouse islet apoptosis and improves engraftment most effectively in the portal and DL subcutaneous sites.
胰岛移植是1型糖尿病和顽固性低血糖的有效治疗方法。然而，仍需要解决移植后胰岛丢失的问题。我们在此探讨泛半胱天冬酶抑制剂F573减轻小鼠门静脉和肝外门静脉部位早期凋亡介导的胰岛死亡的潜力。
将小鼠或人胰岛在含或不含100 μM F573的标准培养基中培养，随后通过末端脱氧核苷酸转移酶dUTP缺口末端标记染色和半胱天冬酶-3激活评估其活力和凋亡情况。将糖尿病小鼠以边缘胰岛剂量（150个胰岛）将同基因胰岛移植到肾被膜（KC）下或皮下无装置（DL）部位，或以全剂量（500个胰岛）移植到门静脉（PV）。将人胰岛以边缘剂量（500个胰岛当量）移植到糖尿病免疫缺陷小鼠的KC下。胰岛在F573存在的情况下培养，并且在移植后第0至5天皮下给予F573。对照小鼠移植未处理的胰岛并注射生理盐水。通过非空腹血糖和葡萄糖耐量试验测量移植物功能。
体外培养后，F573显著降低人和小鼠胰岛凋亡（分别为P < 0.05和P < 0.05）。此外，当移植到KC下时，F573改善了人胰岛功能（P < 0.05）；而F573并未增强小鼠胰岛边缘KC移植。相反，F573显著改善了小鼠胰岛在PV和DL部位的植入（分别为P < 0.05和P < 0.05）。
泛半胱天冬酶抑制剂F573显著降低人和小鼠胰岛凋亡，并在门静脉和DL皮下部位最有效地改善植入。
