Mohammed Arifullah, Chiruvella Kishore K, Rao Yerra Koteswara, Geethangili Madamanchi, Raghavan Sathees C, Ghanta Rama Gopal
Faculty of Agrobased Industry, Universiti Malaysia Kelantan, Jeli Campus, Locked bag-100, 17600, Jeli, Kelantan, Malaysia; Division of Plant Tissue Culture, Department of Botany, Sri Venkateswara University, Tirupati, Andhra Pradesh, India.
Department of Biochemistry, Indian Institute of Science, Bangalore 560 012, India; Division of Plant Tissue Culture, Department of Botany, Sri Venkateswara University, Tirupati, Andhra Pradesh, India.
PLoS One. 2015 Oct 21;10(10):e0141154. doi: 10.1371/journal.pone.0141154. eCollection 2015.
Andrographis lineata is an herbal medicinal plant used in traditional medicine as a substitute for Andrographis paniculata. Here, using mature leaf explants of A. lineata we demonstrate for the first time the callus induction established on MS medium containing 1.0 mg l-1 IAA. Dried callus was subjected to solvent extraction with acetone. Further the acetone residue was separated by silica gel column chromatography, crystallized and characterized on the basis of nuclear magnetic resonance (proton and c13) and liquid chromatographic mass spectroscopy. This analysis revealed the occurrence of two known flavones namely, 7-O-methylwogonin (MW) and Echioidinin (ED). Furthermore, these compounds were tested for their cytotoxicity against leukemic cell line, CEM. We identify that ED and MW induced cytotoxicity in a time- and concentration-dependent manner. Further increase in the LDH release upon treatment with ED and MW further confirmed our cytotoxicity results against leukemic cell line. Strikingly, MW was more potent than ED when compared by trypan blue and MTT assays. Our results recapitulate the utility of callus cultures for the production of plant specific bioactive secondary metabolites instead of using wild plants. Together, our in vitro studies provide new insights of A. lineata callus cultures serving as a source for cancer chemotherapeutic agents.
穿心莲是一种草药植物,在传统医学中用作穿心莲的替代品。在这里,我们首次使用穿心莲的成熟叶片外植体,证明了在含有1.0 mg l-1吲哚乙酸(IAA)的MS培养基上建立愈伤组织诱导。将干燥的愈伤组织用丙酮进行溶剂萃取。进一步通过硅胶柱色谱法分离丙酮残渣,结晶,并基于核磁共振(质子和碳13)以及液相色谱质谱进行表征。该分析揭示了两种已知黄酮的存在,即7-O-甲基汉黄芩素(MW)和紫锥菊黄素(ED)。此外,测试了这些化合物对白血病细胞系CEM的细胞毒性。我们发现ED和MW以时间和浓度依赖性方式诱导细胞毒性。用ED和MW处理后乳酸脱氢酶(LDH)释放的进一步增加进一步证实了我们针对白血病细胞系的细胞毒性结果。令人惊讶的是,通过台盼蓝和MTT试验比较时,MW比ED更有效。我们的结果概括了愈伤组织培养用于生产植物特异性生物活性次生代谢产物而不是使用野生植物的实用性。总之,我们的体外研究为穿心莲愈伤组织培养作为癌症化疗药物来源提供了新的见解。