紫檀花：一种由氧化铜纳米颗粒增强次级代谢物合成的药用宝藏。
摘要
本研究研究了绿色合成的氧化铜纳米颗粒（CuO NPs）在提高毛毛Alhagi（一种具有重要药用价值的药用植物）生物量和治疗性代谢物产量方面的潜力。以毛蕊草叶提取物为还原剂和封盖剂合成CuO NPs，并通过UV-Vis光谱、FTIR、XRD、SEM、TEM和zeta电位分析对其进行了表征。纳米颗粒的大小在7-30纳米之间。在Murashige和Skoog （MS）培养基中添加不同浓度（0 ~ 12 mg/L）的CuO NPs和植物生长调节剂，建立愈伤组织诱导和增殖。在不含CuO NPs的MS培养基中，BAP含量为3.0 mg/L， NAA含量为0.1 mg/L，动蛋白含量为0.50 mg/L时，愈伤组织鲜重最大，子叶为9.02 mg，下胚轴为8.46 mg。然而，CuO NPs以剂量依赖的方式显著增强代谢物的产生。方差分析显示，各生化指标间差异有统计学意义（p=0.001），其中过氧化物酶活性（7755.74）、总黄酮（5195.02）和总可溶性糖（5702.18）的f值较高。在8 mg/L CuO NPs处理下，愈伤组织总游离氨基酸（12.49±0.023 mg/g DW）和总可溶性蛋白（35.617±0.033 mg/g DW）含量升高，淀粉（35.547±0.23 mg/g DW）和总可溶性糖（121.56±0.091 mg/g DW）含量升高。CuO np处理的培养物次生代谢物合成显著增强，在8 mg/L CuO np处理下，总酚类化合物（156.477±0.167 mg/g DW GAE）和总黄酮（58.307±0.179 mg/g QE）含量最高。抗氧化酶分析表明，在特定CuO NP浓度下，子叶愈伤组织表现出最高的抗氧化酶活性：在8 mg/L时，超氧化物歧化酶（抑制84.5±0.254%）和谷胱甘肽还原酶（抑制0.75±0.006%）；过氧化物酶（3.137±0.009 U），过氧化氢酶（77.35±0.152 U），抗坏血酸过氧化物酶（0.43±0.006 mM/mg FW）在10 mg/L。高效液相色谱分析证实，再生根中存在新的抗癌化合物芦皮醇。这些发现表明，CuO NPs有可能提高毛蕊草组织培养中治疗性代谢物的产生，并建议最佳浓度范围（8-10 mg/L）可获得最大效益。需要进一步的研究来阐明控制纳米颗粒与植物相互作用的分子机制，并解决潜在的健康影响。This study investigated the potential of green-synthesized copper oxide nanoparticles (CuO NPs) to enhance biomass production and therapeutic metabolite yields in Alhagi maurorum, a medicinal plant of significant pharmaceutical value. CuO NPs were biosynthesized using A. maurorum leaf extract as a reducing and capping agent, with characterization confirmed via UV-Vis spectroscopy, FTIR, XRD, SEM, TEM, and zeta potential analysis. Nanoparticles ranged from 7-30 nm in size. Callus induction and proliferation were established using Murashige and Skoog (MS) media supplemented with varying concentrations (0-12 mg/L) of CuO NPs combined with plant growth regulators. Maximum callus fresh weight (9.02 mg in cotyledon and 8.46 mg in hypocotyl) was achieved in MS media containing 3.0 mg/L BAP, 0.1 mg/L NAA, and 0.50 mg/L kinetin without CuO NPs. However, CuO NPs significantly enhanced metabolite production in a dose-dependent manner. Analysis of variance revealed statistically significant differences (p=0.001) across all biochemical parameters tested, with high F-values for peroxidase activity (7,755.74), total flavonoids (5,195.02), and total soluble sugar (5,702.18). At 8 mg/L CuO NPs, callus cultures exhibited elevated levels of total free amino acids (12.49±0.023 mg/g DW) and total soluble protein (35.617±0.033 mg/g DW), while control samples produced higher starch (35.547±0.23 mg/g DW) and total soluble sugar (121.56±0.091 mg/g DW) content. Significantly, CuO NP-treated cultures demonstrated enhanced secondary metabolite synthesis, with maximum total phenolic compounds (156.477±0.167 mg/g DW GAE) and flavonoids (58.307±0.179 mg/g QE) at 8 and 10 mg/L CuO NPs, respectively. Antioxidant enzyme analysis revealed that cotyledon-derived callus exhibited peak activities at specific CuO NP concentrations: superoxide dismutase (84.5±0.254% inhibition) and glutathione reductase (0.75±0.006% inhibition) at 8 mg/L; peroxidase (3.137±0.009 U), catalase (77.35±0.152 U), and ascorbate peroxidase (0.43±0.006 mM/mg FW) at 10 mg/L. HPLC analysis confirmed the novel presence of lupeol, an anticancer compound, in regenerated roots. These findings demonstrate the potential of CuO NPs for enhancing therapeutic metabolite production in A. maurorum tissue culture while suggesting optimal concentration ranges (8-10 mg/L) for maximum benefits. Further research is necessary to elucidate the molecular mechanisms governing nanoparticle-plant interactions and to address potential health implications.
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