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过表达葡糖基转移酶基因的转基因亚麻种子的生物医学潜力。

The biomedical potential of genetically modified flax seeds overexpressing the glucosyltransferase gene.

机构信息

Len Pharma, ul, Zamkowa 17, 82-200 Malbork, Poland.

出版信息

BMC Complement Altern Med. 2012 Dec 10;12:251. doi: 10.1186/1472-6882-12-251.

Abstract

BACKGROUND

Flax (Linum usitatissimum) is a potential source of many bioactive components that can be found in its oil and fibers, but also in the seedcake, which is rich in antioxidants. To increase the levels of medically beneficial compounds, a genetically modified flax type (named GT) with an elevated level of phenylopropanoids and their glycoside derivatives was generated. In this study, we investigated the influence of GT seedcake extract preparations on human fibroblast proliferation and migration, and looked at the effect on a human skin model. Moreover, we verified its activity against bacteria of clinical relevance.

METHODS

The GT flax used in this study is characterized by overexpression of the glucosyltransferase gene derived from Solanum sogarandinum. Five GT seedcake preparations were generated. Their composition was assessed using ultra pressure liquid chromatography and confirmed using the UPLC-QTOF method. For the in vitro evaluation, the influence of the GT seedcake preparations on normal human dermal fibroblast proliferation was assessed using the MTT test and the wound scratch assay. A human skin model was used to evaluate the potential for skin irritation. To assess the antimicrobial properties of GT preparations, the percentage of inhibition of bacterial growth was calculated.

RESULTS

The GT seedcake extract had elevated levels of phenylopropanoid compounds in comparison to the control, non-transformed plants. Significant increases in the content of ferulic acid, p-coumaric acid and caffeic acid, and their glucoside derivatives, kaempferol, quercitin and secoisolariciresinol diglucoside (SDG) were observed in the seeds of the modified plants. The GT seedcake preparations were shown to promote the proliferation of normal human dermal fibroblasts and the migration of fibroblasts in the wound scratch assay. The superior effect of GT seedcake extract on fibroblast migration was observed after a 24-hour treatment. The skin irritation test indicated that GT seedcake preparations have no harmful effect on human skin. Moreover, GT seedcake preparations exhibited inhibitory properties toward two bacterial strains: Staphylococcus aureus and Escherichia coli.

CONCLUSIONS

We suggest that preparations derived from the new GT flax are an effective source of phenylopropanoids and that their glycoside derivatives and might be promising natural products with both healing and bacteriostatic effects. This flax-derived product is a good candidate for application in the repair and regeneration of human skin and might also be an alternative to antibiotic therapy for infected wounds.

摘要

背景

亚麻(Linum usitatissimum)是许多生物活性成分的潜在来源,这些成分存在于其油和纤维中,也存在于富含抗氧化剂的种饼中。为了提高具有医学益处的化合物的含量,生成了一种具有高水平苯基丙氨酸及其糖苷衍生物的转基因亚麻(命名为 GT)。在这项研究中,我们研究了 GT 种饼提取物制剂对人成纤维细胞增殖和迁移的影响,并观察了对人体皮肤模型的影响。此外,我们验证了其对临床相关细菌的活性。

方法

本研究中使用的 GT 亚麻的特征是表达来自 Solanum sogarandinum 的葡糖基转移酶基因。生成了五种 GT 种饼制剂。使用超高效液相色谱法评估其组成,并使用 UPLC-QTOF 法进行确认。为了进行体外评估,使用 MTT 试验和划痕试验评估 GT 种饼制剂对正常人类真皮成纤维细胞增殖的影响。使用人体皮肤模型评估皮肤刺激性的潜力。为了评估 GT 制剂的抗菌特性,计算了抑制细菌生长的百分比。

结果

与对照的非转化植物相比,GT 种饼提取物中苯基丙氨酸化合物的含量升高。在转基因植物的种子中观察到阿魏酸、对香豆酸和咖啡酸及其糖苷衍生物、山奈酚、槲皮素和开环异落叶松脂二葡萄糖苷(SDG)的含量显著增加。GT 种饼制剂被证明可促进正常人类真皮成纤维细胞的增殖和划痕试验中成纤维细胞的迁移。在 24 小时处理后,观察到 GT 种饼提取物对成纤维细胞迁移的促进作用更为明显。皮肤刺激试验表明,GT 种饼制剂对人体皮肤没有有害影响。此外,GT 种饼制剂对两种细菌菌株(金黄色葡萄球菌和大肠杆菌)表现出抑制作用。

结论

我们建议,来自新型 GT 亚麻的制剂是苯基丙氨酸及其糖苷衍生物的有效来源,其糖苷衍生物可能是具有愈合和抑菌作用的有前途的天然产物。这种源自亚麻的产品是修复和再生人体皮肤的候选药物,也可能是感染伤口抗生素治疗的替代方法。

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