Pharmacological Institute of New Drugs, School of Pharmacy, Shandong University, Ji'nan, Shandong 250012, P.R. China.
Brain Res. 2011 Jan 12;1368:1-10. doi: 10.1016/j.brainres.2010.10.064. Epub 2010 Oct 23.
Low-molecular-weight heparin (LMWH) and ultra-low-molecular-weight heparin (ULMWH) are heparin's derivatives, having various pharmacological effects. The present study aims to investigate the effect of ULMWH on amyloid β peptide (Aβ25-35)-induced neurotoxicity in cultured rat cortical neurons, and LMWH was employed as a positive control agent. The neurons were incubated with Aβ25-35 (35μM), Aβ25-35 plus ULMWH (2, 10, 50 μg/ml) or LMWH (10 μg/ml) for 24h. The cell viability was assessed by MTT and LDH release. FITC-Annexin V/PI double staining, Hoechst 33258 staining, TUNEL and Western blotting for bcl-2 and caspase-3 were employed to measure the neuron apoptosis. Furthermore, the intracellular Ca(2+) concentration was measured by a fluorescent dye, Fura-2/AM. The results showed that ULMWH significantly increased cell viability and the protein expression levels of bcl-2 and decreased the LDH release, the number of apoptotic cells, the concentration of intracellular Ca(2+) and the protein expression levels of caspase-3 in cortical neurons, suggesting that ULMWH can obviously reduce Aβ25-35-induced neurotoxic effects and might act as a potential agent for Alzheimer's disease.
低分子量肝素 (LMWH) 和超低分子量肝素 (ULMWH) 是肝素的衍生物，具有多种药理学作用。本研究旨在探讨 ULMWH 对培养的大鼠皮质神经元中淀粉样 β 肽 (Aβ25-35) 诱导的神经毒性的影响，并将 LMWH 用作阳性对照剂。将神经元与 Aβ25-35（35μM）、Aβ25-35 加 ULMWH（2、10、50μg/ml）或 LMWH（10μg/ml）孵育 24 小时。通过 MTT 和 LDH 释放评估细胞活力。使用 FITC-Annexin V/PI 双重染色、Hoechst 33258 染色、TUNEL 和 Western blot 检测 bcl-2 和 caspase-3，以测量神经元凋亡。此外，通过荧光染料 Fura-2/AM 测量细胞内 Ca(2+)浓度。结果表明，ULMWH 可显著增加皮质神经元的细胞活力和 bcl-2 蛋白表达水平，降低 LDH 释放、凋亡细胞数量、细胞内 Ca(2+)浓度和 caspase-3 蛋白表达水平，提示 ULMWH 可明显减轻 Aβ25-35 诱导的神经毒性作用，可能成为阿尔茨海默病的潜在治疗药物。
