Introduction

Adaptogens are natural compounds or plant extracts that increase adaptability and survival of organisms under stress. Adaptogens stimulate cellular and organismal defense systems by activating intracellular and extracellular signaling pathways and expression of stress-activated proteins and neuropeptides. The effects adaptogens on mediators of adaptive stress response and longevity signaling pathways have been reported, but their stress-protective mechanisms are still not fully understood.

Aim of the study

The aim of this study was to identify key molecular mechanisms of adaptogenic plants traditionally used to treat stress and aging-related disorders, i.e., Rhodiola rosea, Eleutherococcus senticosus, Withania somnifera, Rhaponticum carthamoides, and Bryonia alba.

Materials and methods

To investigate the underlying molecular mechanisms of adaptogens, we conducted RNA sequencing to profile gene expression alterations in T98G neuroglia cells upon treatment of adaptogens and analyzed the relevance of deregulated genes to adaptive stress-response signaling pathways using in silico pathway analysis software.

Results and discussion

At least 88 of the 3516 genes regulated by adaptogens were closely associated with adaptive stress response and adaptive stress-response signaling pathways (ASRSPs), including neuronal signaling related to corticotropin-releasing hormone, cAMP-mediated, protein kinase A, and CREB; pathways related to signaling involving CXCR4, melatonin, nitric oxide synthase, GP6, Gαs, MAPK, neuroinflammation, neuropathic pain, opioids, renin–angiotensin, AMPK, calcium, and synapses; and pathways associated with dendritic cell maturation and G-coupled protein receptor–mediated nutrient sensing in enteroendocrine cells. All samples tested showed significant effects on the expression of genes encoding neurohormones CRH, GNRH, UCN, G-protein–coupled and other transmembrane receptors TLR9, PRLR, CHRNE, GP1BA, PLXNA4, a ligand-dependent nuclear receptor RORA, transmembrane channels, transcription regulators FOS, FOXO6, SCX, STAT5A, ZFPM2, ZNF396, ZNF467, protein kinases MAPK10, MAPK13, MERTK, FLT1, PRKCH, ROS1, TTN), phosphatases PTPRD, PTPRR, peptidases, metabolic enzymes, a chaperone (HSPA6), and other proteins, all of which modulate numerous life processes, playing key roles in several canonical pathways involved in defense response and regulation of homeostasis in organisms. It is for the first time we report that the molecular mechanism of actions of melatonin and plant adaptogens are alike, all adaptogens tested activated the melatonin signaling pathway by acting through two G-protein–coupled membrane receptors MT1 and MT2 and upregulation of the ligand-specific nuclear receptor RORA, which plays a role in intellectual disability, neurological disorders, retinopathy, hypertension, dyslipidemia, and cancer, which are common in aging. Furthermore, melatonin activated adaptive signaling pathways and upregulated expression of UCN, GNRH1, TLR9, GP1BA, PLXNA4, CHRM4, GPR19, VIPR2, RORA, STAT5A, ZFPM2, ZNF396, FLT1, MAPK10, MERTK, PRKCH, and TTN, which were commonly regulated by all adaptogens tested. We conclude that melatonin is an adaptation hormone playing an important role in regulation of homeostasis. Adaptogens presumably worked as eustressors (“stress-vaccines”) to activate the cellular adaptive system by inducing the expression of ASRSPs, which then reciprocally protected cells from damage caused by distress. Functional investigation by interactive pathways analysis demonstrated that adaptogens activated ASRSPs associated with stress-induced and aging-related disorders such as chronic inflammation, cardiovascular health, neurodegenerative cognitive impairment, metabolic disorders, and cancer.

Conclusion

This study has elucidated the genome-wide effects of several adaptogenic herbal extracts in brain cells culture. These data highlight the consistent activation of ASRSPs by adaptogens in T98G neuroglia cells. The extracts affected many genes playing key roles in modulation of adaptive homeostasis, indicating their ability to modify gene expression to prevent stress-induced and aging-related disorders. Overall, this study provides a comprehensive look at the molecular mechanisms by which adaptogens exerts stress-protective effects.

中文翻译：

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利用系统生物学研究草药提取物对离体脑细胞适应性作用的新分子机制

介绍

适应原是天然化合物或植物提取物，可提高生物在压力下的适应性和存活率。适应原通过激活细胞内和细胞外信号通路以及应激激活的蛋白质和神经肽的表达来刺激细胞和机体防御系统。已经报道了适应原对适应性应激反应和长寿信号传导途径的介体的作用，但是它们的应激保护机制仍未被完全理解。

研究目的

这项研究的目的是确定传统上用于治疗压力和与衰老相关的疾病的适应性植物的关键分子机制，即玫瑰红景天，刺五加肠球菌，Withania somnifera，Rhaponticum carhamoides和Bryonia alba。

材料和方法

为了研究适应原的潜在分子机制，我们进行了RNA测序，分析了适应原处理后T98G神经胶质细胞中基因表达的变化，并使用计算机模拟分析软件分析了失调基因与适应性应激反应信号通路的相关性。

结果与讨论

适应原调节的3516个基因中，至少有88个与适应性应激反应和适应性应激反应信号通路（ASRSPs）密切相关，包括与促肾上腺皮质激素释放激素，cAMP介导的蛋白激酶A和CREB相关的神经元信号。与CXCR4，褪黑素，一氧化氮合酶，GP6，Gαs，MAPK，神经发炎，神经性疼痛，阿片类药物，肾素-血管紧张素，AMPK，钙和突触相关的信号通路；肠内分泌细胞中与树突状细胞成熟和G偶联蛋白受体介导的营养感测相关的途径。所有测试样品均显示出对编码神经激素CRH，GNRH，UCN，G蛋白偶联和其他跨膜受体TLR9，PRLR，CHRNE，GP1BA，PLXNA4，配体依赖性核受体RORA的基因表达有显着影响。跨膜通道，转录调节因子FOS，FOXO6，SCX，STAT5A，ZFPM2，ZNF396，ZNF467，蛋白激酶MAPK10，MAPK13，MERTK，FLT1，PRKCH，ROS1，TTN），磷酸酶PTPRD，PTPRR，肽酶，代谢酶，伴侣蛋白H（伴侣蛋白H ）和其他蛋白质，所有这些蛋白质均可调节许多生命过程，在生物体的防御反应和体内稳态调节所涉及的几种规范途径中起着关键作用。这是我们首次报道褪黑激素和植物适应原的作用的分子机制是相似的，所有测试的适应原均通过两个G蛋白偶联的膜受体MT1和MT2以及配体上调而激活了褪黑素信号传导途径。特定的核受体RORA，在智力障碍，神经系统疾病，视网膜病变，高血压，血脂异常，和癌症，这在衰老中很常见。此外，褪黑素激活了普遍调节的UCN，GNRH1，TLR9，GP1BA，PLXNA4，CHRM4，GPR19，VIPR2，RORA，STAT5A，ZFPM2，ZNF396，FLT1，MAPK10，MERTK，PRKCH和TTN的适应性信号通路并上调表达经所有适应原测试。我们得出的结论是，褪黑激素是一种适应激素，在体内稳态调节中起着重要作用。适应原大概是通过诱导ASRSPs的表达来激活细胞适应性系统，从而起到了胁迫的作用（“应激疫苗”），然后ASRSPs可以相应地保护细胞免受由窘迫引起的损害。通过互动途径分析进行的功能研究表明，适应原激活了ASRSP，这些ASRSP与压力引起的和衰老相关的疾病（例如慢性炎症，心血管健康，

结论

这项研究阐明了几种适应性强的草药提取物在脑细胞培养中的全基因组作用。这些数据突显了T98G神经胶质细胞中的适应原激活了ASRSPs。提取物影响了许多基因，这些基因在调节适应性稳态中起着关键作用，表明它们具有修饰基因表达以预防压力引起的衰老相关疾病的能力。总的来说，这项研究全面介绍了适应原发挥应力保护作用的分子机制。