Kirakosyan Rima N, Kalasnikova Elena A, Bolotina Elizaveta A, Saleh Abdulrahman, Balakina Anastasiya A, Zaytseva Svetlana M
Department of Biotechnology, Russian State Agrarian University-Moscow Timiryazev Agricultural Academy, Timiryazevskaya Street 49, Moscow 127434, Russia.
Federal Research Center of Problems of Chemical Physics and Medicinal Chemistry, Russian Academy of Science, Ac. Semenov Avenue 1, Moscow Region, Chernogolovka, Moscow 142432, Russia.
Life (Basel). 2024 Dec 20;14(12):1694. doi: 10.3390/life14121694.
In order to scientifically search for new sources of secondary metabolites with valuable qualities for phytopharmacognosy, tasks requiring a step-by-step solution were set. The primary task is the development of technologies for obtaining in vitro highly productive biomass of cells of relict gymnosperms of the genus , capable of accumulating various classes of secondary metabolites. The study of the accumulation and localization of secondary metabolites allowed us to evaluate the biological activity and cytotoxicity of in vitro cultures. In our study, histochemical methods were used to determine the localization of secondary compounds (phenolic and terpenoid in nature) in plant tissues. Secondary metabolites-polyphenols, catechins, and terpenoids-are mainly localized in the epidermal, parenchymal, and conductive tissues of leaves and stems. In callus and suspension cultures of , secondary metabolites were localized in cell walls and vacuoles. The mineral composition of the nutrient medium (MS and WPM), the light source (photoperiod), and the endogenous content of polyphenols in the primary explant influenced the initiation and growth characteristics of the in vitro culture of plants. Inhibition of growth in suspension cultures on the WPM nutrient medium was noted. The cultivation of cell lines at a 16 h photoperiod stimulated the formation of polyphenols but had a negative effect on the growth of callus cultures. Extractive substances obtained from intact and callus tissues of evergreen demonstrate high biological (fungicidal) activity and cytotoxicity. The inhibitory effect on was noted when 200 mg/L of extract was added to the nutrient medium. Extracts of redwood callus cultures were low in toxicity to normal FetMSC cells but inhibited the growth of lines of "immortal" cervical HeLa cancer cells and human glioblastoma A172. Intact tissues of plants and cell cultures initiated from them in vitro are producers of secondary metabolites with high biological activity.
为了科学地寻找具有药用植物学珍贵品质的次生代谢物新来源，设定了需要逐步解决的任务。首要任务是开发技术，以获得能够积累各类次生代谢物的该属残遗裸子植物细胞的体外高产生物质。对次生代谢物积累和定位的研究使我们能够评估体外培养物的生物活性和细胞毒性。在我们的研究中，采用组织化学方法确定植物组织中次生化合物（天然酚类和萜类）的定位。次生代谢物——多酚、儿茶素和萜类——主要定位于叶和茎的表皮、薄壁和传导组织中。在该植物的愈伤组织和悬浮培养物中，次生代谢物定位于细胞壁和液泡中。营养培养基（MS和WPM）的矿物质组成、光源（光周期）以及初代外植体中多酚的内源含量影响该植物体外培养的起始和生长特性。注意到在WPM营养培养基上悬浮培养物的生长受到抑制。在16小时光周期下培养该细胞系刺激了多酚的形成，但对愈伤组织培养物的生长有负面影响。从常绿该植物的完整组织和愈伤组织中获得的提取物具有高生物（杀真菌）活性和细胞毒性。当向营养培养基中添加200mg/L的该植物提取物时，观察到对（某种生物）的抑制作用。红木愈伤组织培养物的提取物对正常的人胎儿间充质干细胞（FetMSC）细胞毒性较低，但抑制了“永生”宫颈癌细胞系HeLa和人胶质母细胞瘤细胞系A172的生长。该植物的完整组织及其体外诱导的细胞培养物是具有高生物活性的次生代谢物的生产者。
