Department of Biotechnology, Guru Nanak Dev University, Amritsar, Punjab, India.
Amity Institute of Biotechnology, Amity University, Haryana, India.
PLoS One. 2020 Mar 25;15(3):e0230142. doi: 10.1371/journal.pone.0230142. eCollection 2020.
Neuroinflammation is a major risk factor associated with the pathogenesis of neurodegenerative diseases. Conventional non-steroidal anti-inflammatory drugs are prescribed but their long term use is associated with adverse effects. Thus, herbal based medicines are attracting major attraction worldwide as potential therapeutic candidates. Tylophora indica (Burm. f) Merrill is a valuable medicinal plant well known in Ayurvedic practices for its immunomodulatory, anti-oxidant, anti-asthmatic and antirheumatic activities. The present study aimed to elucidate the anti-neuroinflammatory potential of water and hydroalcoholic leaf extracts of micropropagated plants of T. indica using BV-2 microglia activated with lipopolysaccharide as an in vitro model system and development of an efficient reproducible protocol for its in vitro cloning. Non cytotoxic doses of the water and hydroalcoholic extracts (0.2μg/ml and 20μg/ml, respectively) were selected using MTT assay. α-Tubulin, Iba-1 and inflammatory cascade proteins like NFκB, AP1 expression was studied using immunostaining to ascertain the anti-neuroinflammatory potential of these extracts. Further, anti-migratory activity was also analyzed by Wound Scratch Assay. Both extracts effectively attenuated lipopolysaccharide induced microglial activation, migration and the production of nitrite via regulation of the expression of NFκB and AP1 as the possible underlying target molecules. An efficient and reproducible protocol for in vitro cloning of T. indica through multiple shoot proliferation from nodal segments was established on both solid and liquid Murashige and Skoog's (MS) media supplemented with 15μM and 10μM of Benzyl Amino Purine respectively. Regenerated shoots were rooted on both solid and liquid MS media supplemented with Indole-3-butyric acid (5-15μM) and the rooted plantlets were successfully acclimatized and transferred to open field conditions showing 90% survivability. The present study suggests that T. indica may prove to be a potential anti-neuroinflammatory agent and may be further explored as a potential therapeutic candidate for the management of neurodegenerative diseases. Further, the current study will expedite the conservation of T. indica ensuring ample supply of this threatened medicinal plant to fulfill its increasing demand in herbal industry.
神经炎症是与神经退行性疾病发病机制相关的主要风险因素。目前常处方使用非甾体抗炎药，但长期使用这些药物与不良反应相关。因此，基于草药的药物作为潜在的治疗候选药物引起了全世界的关注。印度娃儿藤（Burm. f）Merrill 是一种有价值的药用植物，在阿育吠陀医学实践中因其具有免疫调节、抗氧化、抗哮喘和抗风湿作用而闻名。本研究旨在通过使用脂多糖激活的 BV-2 小胶质细胞作为体外模型系统，阐明微繁殖印度娃儿藤的水和水醇叶提取物的抗神经炎症潜力，并开发一种有效的、可重复的体外克隆方法。使用 MTT 测定法选择水和水醇提取物的非细胞毒性剂量（分别为 0.2μg/ml 和 20μg/ml）。使用免疫染色研究 α-微管蛋白、Iba-1 和炎症级联蛋白（如 NFκB、AP1）的表达，以确定这些提取物的抗神经炎症潜力。此外，还通过划痕实验分析了抗迁移活性。这两种提取物都通过调节 NFκB 和 AP1 的表达，有效地减轻了脂多糖诱导的小胶质细胞激活、迁移和亚硝酸盐的产生，这可能是潜在的作用靶点。在固体和液体 Murashige 和 Skoog（MS）培养基上，通过节间丛生增殖建立了印度娃儿藤的有效和可重复的体外克隆方法，培养基中分别添加了 15μM 和 10μM 的苄基氨基嘌呤。再生芽在固体和液体 MS 培养基上生根，培养基中添加了吲哚-3-丁酸（5-15μM），生根的幼苗成功适应并转移到开放田间条件下，存活率达到 90%。本研究表明，印度娃儿藤可能是一种有潜力的抗神经炎症药物，并可能进一步探索作为神经退行性疾病管理的潜在治疗候选药物。此外，本研究将加速印度娃儿藤的保护，以确保这种受到威胁的药用植物有足够的供应，以满足其在草药行业中不断增长的需求。
