Li Jiejing, Li Yanxi, Zhang Penghui, Niu Hua, Shi Yu
Department of Clinical Laboratory, Children's Hospital of Chongqing Medical University, Chongqing 400014, P.R. China.
Laboratory of Developmental Diseases in Childhood of Education Ministry, Key Laboratory of Pediatrics in Chongqing, Chongqing International Science and Technology Cooperation Center for Child Development and Disorder, Children's Hospital of Chongqing Medical University, Chongqing 400014, P.R. China.
Mol Med Rep. 2014 Dec;10(6):2805-10. doi: 10.3892/mmr.2014.2576. Epub 2014 Sep 18.
Nicotinic acid (NA), a member of the vitamin B family, is well known for its functions in the treatment and prevention of atherosclerosis due to decreasing plasma levels of low-density lipoprotein cholesterol. In recent years, the major side effect of NA, cutaneous flushing, has also attracted extensive attention. However, the effects of NA in other aspects of physiology or cell biology have remained elusive. The present study provided evidence that high concentrations of NA were able to first reduce and later elevate intracellular [Ca2+] in the NIH3T3 cell line. The reduction of the intracellular Ca2+ concentration was achieved within the initial 10 sec, and was preceded by a gradual elevation of intracellular [Ca2+]. Notably, marked accumulation of opaque materials in the perinuclear region was observed in NIH3T3 cells treated with 70 mM NA. Further analysis revealed that treatment with 70 mM NA for 1 h disassembled the microtubule and F‑actin cytoskeleton systems and resulted in β‑tubulin degradation in an ubiquitin‑proteasome-dependent manner. These data indicated that high concentrations of NA disrupted cytoskeleton structures, which may have contributed to minus end (nucleus region) to plus end (cell membrane region)-directed transport processes and resulted in the deposition of material in the perinuclear region. Artificially increasing [Ca2+] adding CaCl2 to the culture media effected the disassembly of F‑actin, while it had no apparent effect on microtubules. These results suggested that the disruption of the cytoskeleton systems was not entirely due to the NA-induced elevation of [Ca2+]. Finally, microinjection of NA into xenopus embryos blocked the transport of melanosomes to the peripheral cellular area. In conclusion, the present study indicated that NA disassembles F‑actin and microtubule systems, thereby blocking cytoskeleton-dependent intracellular transport.
烟酸(NA)是维生素B族的一员,因其在降低血浆低密度脂蛋白胆固醇水平从而治疗和预防动脉粥样硬化方面的作用而广为人知。近年来,NA的主要副作用——皮肤潮红,也引起了广泛关注。然而,NA在生理或细胞生物学其他方面的作用仍不明确。本研究提供了证据表明,高浓度的NA能够先降低然后升高NIH3T3细胞系中的细胞内[Ca2+]。细胞内Ca2+浓度的降低在最初10秒内实现,并且在此之前细胞内[Ca2+]有逐渐升高的过程。值得注意的是,在用70 mM NA处理的NIH3T3细胞中观察到核周区域有明显的不透明物质积累。进一步分析表明,用70 mM NA处理1小时会破坏微管和F-肌动蛋白细胞骨架系统,并以泛素-蛋白酶体依赖的方式导致β-微管蛋白降解。这些数据表明,高浓度的NA破坏了细胞骨架结构,这可能导致了从负端(核区域)到正端(细胞膜区域)的定向运输过程,并导致物质在核周区域沉积。通过向培养基中添加CaCl2人为增加[Ca2+]会影响F-肌动蛋白的解聚,而对微管没有明显影响。这些结果表明,细胞骨架系统的破坏并不完全是由于NA诱导的[Ca2+]升高。最后,将NA显微注射到非洲爪蟾胚胎中会阻止黑素小体向细胞周边区域的运输。总之,本研究表明NA会破坏F-肌动蛋白和微管系统,从而阻断依赖细胞骨架的细胞内运输。