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Abstract：
The dried bark of Phellodendron chinense has been used as a traditional herbal medicine to remove damp heat, relieve consumptive fever, and cure dysentery and diarrhea. In the present study, we performed quantitative analyses of the two components of P. chinense, phellodendrine and berberine, using high-performance liquid chromatography. A 70% ethanol extract of P. chinense was prepared and the two components were separated on a C-18 analytical column using a gradient solvent system of acetonitrile and 0.1% (v/v) aqueous trifluoroacetic acid. The ultraviolet wavelength used for detection was 200 nm for phellodendrine and 226 nm for berberine. The analytical method established here showed high linearity (correlation coefficient, ≥0.9991). The amount of phellodendrine and berberine used was 22.255 ± 0.123 mg/g and 269.651 ± 1.257 mg/g, respectively. Moreover, we performed an in vitro acetylcholinesterase (AChE) activity assay and an amyloid-β aggregation test to examine the biological properties of phellodendrine and berberine as therapeutic drugs for Alzheimer\'s disease. Phellodendrine and berberine inhibited AChE activity in a dose-dependent manner (IC50 = 36.51 and 0.44 μM, respectively). In contrast, neither phellodendrine nor berberine had an effect on amyloid-β aggregation. The P. chinense extract and phellodendrine, but not berberine, exhibited antioxidant activity by increasing radical scavenging activity. Moreover, P. chinense demonstrated a neuroprotective effect in hydrogen peroxide-treated HT22 hippocampal cells. Overall, our findings suggest that P. chinense has potential as an anti-Alzheimer\'s agent via the suppression of the enzymatic activity of acetylcholinesterase and the stimulation of antioxidant activity.
摘要：
黄柏的干燥树皮已被用作传统的草药来消除湿热，缓解消耗性发烧，治疗痢疾和腹泻。在本研究中,我们对P.chinense的两种成分进行了定量分析，黄柏碱和小檗碱,采用高效液相色谱法。制备P.chinense的70％乙醇提取物，并使用乙腈和0.1％（v/v）三氟乙酸水溶液的梯度溶剂系统在C-18分析柱上分离两种成分。用于检测的紫外线波长为黄柏碱200nm，小檗碱226nm。此处建立的分析方法显示出高线性(相关系数，≥0.9991)。黄柏碱和小檗碱的用量分别为22.255±0.123mg/g和269.651±1.257mg/g,分别。此外，我们进行了体外乙酰胆碱酯酶（AChE）活性测定和淀粉样蛋白-β聚集试验，以检查黄柏碱和小檗碱作为阿尔茨海默病治疗药物的生物学特性。黄柏碱和黄连素以剂量依赖性方式抑制AChE活性（IC50=36.51和0.44μM，分别)。相比之下,黄柏碱和小檗碱对β淀粉样蛋白聚集均无影响。黄柏提取物和黄柏碱，但不是小檗碱,通过增加自由基清除活性而表现出抗氧化活性。此外，P.chinense在过氧化氢处理的HT22海马细胞中表现出神经保护作用。总的来说,我们的研究结果表明，中国山梨可能通过抑制乙酰胆碱酯酶的酶活性和刺激抗氧化活性作为抗阿尔茨海默病的药物。
