1,25-(OH)2D3 protects Schwann cells against advanced glycation end products-induced apoptosis through PKA-NF-κB pathway
1,25-(OH)2D3 protects Schwann cells against advanced glycation end products-induced apoptosis through PKA-NF-κB pathway
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1,25-(OH)(2)D-3 通过 PKA-NF-kappa B 途径保护雪旺细胞免受晚期糖基化终产物诱导的细胞凋亡
DOI：
10.1016/j.lfs.2019.03.068
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发表时间：
2019-05-15
期刊：
影响因子：
6.1
通讯作者：
Peng, Liang
中科院分区：
文献类型：
--
作者：
Xu, Shiqing;Li, Jing;Peng, Liang
目的：探讨1，25-(OH)(2)D-3对晚期糖基化终产物诱导雪旺细胞凋亡的影响及其机制。主要方法：用AGE修饰的牛血清白蛋白(AGE)孵育大鼠坐骨神经雪旺细胞，以1，25-(OH)(2)D-3为保护剂。PI/Annexin-V染色、caspase3活性测定和caspase3、PARP蛋白免疫印迹法检测细胞凋亡。免疫印迹法检测蛋白激酶A(PKA)和核因子-kappaB(NF-kappa B)的活性。免疫荧光染色用于核因子-kappaB的细胞间定位，酶联免疫吸附分析法检测细胞因子的分泌。主要发现：雪旺细胞与AGE孵育后，细胞凋亡加速。然而，如果将1，25(OH)(2)D-3与AGE结合，细胞凋亡率显著减少。1，25-(OH)(2)D-3增强PKA活性，但抑制AGE诱导的NF-kappa B的核转位。此外，PKA激活剂(8-溴腺苷环腺苷单磷酸，8-BrcAMP)或NF-kappa B抑制剂(咖啡酸苯乙酯，CAPE)可减少细胞的凋亡，减少caspase 3的裂解和PARP的裂解，提示1，25-(OH)(2)D-3对雪旺细胞的保护作用与PKA和NF-kappa B通路有关。8-溴-cAMP和CAPE可抑制AGE诱导的白细胞介素1β、前列腺素E_2和环氧合酶2的分泌。有趣的是，8-溴-cAMP降低了磷酸化的核因子-kappaB，抑制了核因子-kappaB的核转位，暗示了PKA对NFKB的调节。H-89(PKA抑制剂)可阻断1，25-(OH)(2)D-3对细胞凋亡的保护作用。结论：1，25-(OH)(2)D-3可通过PKA/NF-kappa B途径保护雪旺细胞免受AGE诱导的细胞凋亡。意义：这些发现为维生素D治疗糖尿病神经病变提供了实验依据。
Aims: To explore the effect and mechanism of 1, 25-(OH)(2)D-3 on Schwann cell apoptosis induced by advanced glycation end products.Main methods: Schwann cells, isolated from rodent sciatic nerve were incubated with AGE-modified bovine serum albumin(AGE) to mimic diabetic conditions and 1,25-(OH)(2)D-3 was used as protector. Cell apoptosis was detected by PI/Annexin-V staining, caspase 3 activity assay and western blotting for caspase 3 and PARP. The activation of protein kinase A (PKA) and nuclear factor kappa-B (NF-kappa B) was evaluated by western blot. Immunofluorescent staining was used for intercellular location of NF-kappa B. Cytokine secretion was evaluated by enzyme-linked immunosorbent assay.Key findings: Schwann cell apoptosis accelerated after incubating with AGE. However, if combining 1,25(OH)(2)D-3 with AGE, apoptosis decreased significantly. 1,25-(OH)(2)D-3 enhanced PKA activity, but inhibited AGE-induced nuclear translocation of NF-kappa B. Furthermore, PKA activator (8-bromoadenoside cyclic adenoside monophosphate, 8-Br-cAMP) or NF-kappa B inhibitor (caffeic acid phenethyl ester, CAPE) could reduce the apoptosis, decreased cleaved caspase 3 and cleaved PARP, suggesting the involvement of PKA and NF-kappa B pathways in the protection of 1,25-(OH)(2)D-3 on Schwann cells. Moreover, 8-Br-cAMP and CAPE could inhibit AGE-induced secretion of interleukin(IL)-1 beta, prostaglandin E2(PEG2) and cyclooxygenase 2(COX2). Interestingly, 8-Br-cAMP decreased phospho-NF-kappa B and inhibited nucleus translocation of NF-kappa B. It hinted at the regulation of PKA to NFKB. Finally, a pre-treatment of H-89 (an inhibitor of PKA) could block the protection of 1,25-(OH)(2)D-3 on cell apoptosis. In conclusion, 1,25-(OH)(2)D-3 could protect Schwann cell against AGE-induced apoptosis through PKA/NF-kappa B pathway.Significance: These findings provide experimental rationales for using vitamin D for diabetic neuropathy.