Gynecology Research Unit, Hannover Medical School, Carl-Neuberg-Strasse 1, D-30625 Hannover, Germany.
Department of Obstetrics and Gynecology, Hannover Medical School, Carl-Neuberg-Strasse 1, D-30625 Hannover, Germany.
Int J Mol Sci. 2021 Sep 8;22(18):9696. doi: 10.3390/ijms22189696.
Immunosuppressants are a mandatory therapy for transplant patients to avoid rejection of the transplanted organ by the immune system. However, there are several known side effects, including alterations of the vasculature, which involve a higher occurrence of cardiovascular events. While the effects of the commonly applied immunosuppressive drugs cyclosporine A (CsA) and tacrolimus (Tac) on mature endothelial cells have been addressed in several studies, we focused our research on the unexplored effects of CsA and Tac on endothelial colony-forming cells (ECFCs), a subgroup of endothelial progenitor cells, which play an important role in vascular repair and angiogenesis. We hypothesized that CsA and Tac induce functional defects and activate an inflammatory cascade via NF-κB signaling in ECFCs. ECFCs were incubated with different doses (0.01 µM-10 µM) of CsA or Tac. ECFC function was determined using in vitro models. The expression of inflammatory cytokines and adhesion molecules was explored by quantitative real-time PCR and flow cytometry. NF-κB subunit modification was assessed by immunoblot and immunofluorescence. CsA and Tac significantly impaired ECFC function, including proliferation, migration, and tube formation. TNF-α, IL-6, VCAM, and ICAM mRNA expression, as well as PECAM and VCAM surface expression, were enhanced. Furthermore, CsA and Tac led to NF-κB p65 subunit phosphorylation and nuclear translocation. Pharmacological inhibition of NF-κB by parthenolide diminished CsA- and Tac-mediated proinflammatory effects. The data of functional impairment and activation of inflammatory signals provide new insight into mechanisms associated with CsA and Tac and cardiovascular risk in transplant patients.
免疫抑制剂是移植患者的强制性治疗方法，以避免免疫系统对移植器官的排斥。然而，有几种已知的副作用，包括血管改变，这涉及更高的心血管事件发生率。虽然几种研究已经探讨了常用免疫抑制剂环孢素 A（CsA）和他克莫司（Tac）对成熟内皮细胞的影响，但我们的研究重点是 CsA 和 Tac 对内皮祖细胞（EPC）的一个亚群——内皮集落形成细胞（ECFC）的未探索作用，后者在血管修复和血管生成中发挥重要作用。我们假设 CsA 和 Tac 通过 NF-κB 信号通路在 ECFC 中诱导功能缺陷和激活炎症级联反应。用不同剂量（0.01 µM-10 µM）的 CsA 或 Tac 孵育 ECFC。通过体外模型测定 ECFC 功能。通过定量实时 PCR 和流式细胞术研究炎症细胞因子和粘附分子的表达。通过免疫印迹和免疫荧光评估 NF-κB 亚基修饰。CsA 和 Tac 显著损害 ECFC 功能，包括增殖、迁移和管形成。TNF-α、IL-6、VCAM 和 ICAM mRNA 表达以及 PECAM 和 VCAM 表面表达增强。此外，CsA 和 Tac 导致 NF-κB p65 亚基磷酸化和核转位。用小白菊内酯抑制 NF-κB 可减少 CsA 和 Tac 介导的促炎作用。功能障碍和炎症信号激活的数据为与 CsA 和 Tac 相关的机制以及移植患者的心血管风险提供了新的见解。
