Unveiling novel longevity-regulating mechanisms of phytogenic adaptogen Phytocee^™ through a computational biology approach

KShamana1

EdwinR1

JaishreeSP1

DPrashanth1

GAboli1✉

MDeepak1

AboliGirme2Emailaboli@anturalrmedy.com

1Natural Remedies Private Limited560091BangaloreIndia

2Natural Remedies Private LimitedBangaloreIndia, Pin Code=560100

Shamana K, Edwin R, Jaishree SP, Prashanth D, Aboli G*, Deepak M

Natural Remedies Private Limited, Bangalore, India 560091

*Corresponding Authors:

Dr Aboli Girme

aboli@anturalrmedy.com

Natural Remedies Private Limited,

Bangalore, India, Pin Code = 560100

Abstract

Phytogenics is a promising and novel sector in animal health. Indian botanicals have been shown to protect animals from various diseases and health stressors, which include nutritional, environmental, biological, and chemical stressors. These stressors may cause sudden mortality in the animals and are a matter of concern among farmers, which Phytocee^™ can address. It consists of scientifically validated (LC-MS/MS) phytochemicals belonging to the classes of triterpenoids, hydrolysable tannins, and withanolides that help enhance the adaptogenic potential of the animals. The adaptogenic potential of an animal is associated with prolonging their lifespan. This research investigated the pathway-focused approaches for increasing longevity in animals at a cellular level by observing their differentially expressed genes (DEGs): FOXO, AMPK, SIRT1, and mTOR using computational tools for gene enrichment: DIGEP Pred2.0, which showed significant modulation of the DEGs (1.25e-159), especially for the enriched term “longevity regulating pathway”. Pathway mapping using SHINYGO0.82, KEGG, showed significant mapping of these genes in the enriched pathway. Network pharmacology was done to detect protein-protein interactions for these DEGs using INPUT2.0. The heat maps and flowerpot Venn diagrams were generated for enriched pathways using SR-PLOT, which showed that the maximum genes were responsible for regulating the longevity of the animals. The study concluded with a possible molecular pharmacology of Phytocee^TM, suggesting that the product may have helped overcome stressors by increasing the lifespan of various animal species, including poultry, swine, and aquatic animals. The possibility can be further validated through the wet lab trials.

Graphical Abstract

Keywords:

Longevity regulating pathway

Adaptogenic potential

Phytocee™

Introduction

The global health market sees great potential in the adaptogen market. In the supplement industry, a shift toward sustainable animal production is increasing demand for herbal products over chemical ones. Many botanicals such as Emblica officinalis (Amla), Ocimum sanctum (Tulsi) and Withania sominifera (Ashwagandha), Bacopa monneiri (Brahmi), Tinospora cordifolia (Guduchi) and Glycyrrhiza glabra (Licorice) have been shown to have excellent resistance to different kinds of stressors and improve animal health. These adaptogenic plant-based products and their phytochemicals have been a popular and effective choice in animal food and feed supplements. [1–4]

Adaptogens play a vital role in animal health by reducing multiple stresses during their life cycle. The various types of stressors in animals can be attributed to nutritional deficiencies or compromised management systems on farms, which may include inadequate ventilation, an ammoniacal smell in the environment, or internal stressors such as a lack of immunity to infectious diseases and susceptibility to thermogenic stress. Due to all these stressors, there is a heavy risk of high incidences of mortality, that are a cause of great concern in industry. Enhancing the adaptogenic potential of the animals is the pressing need to alleviate stress levels, thereby reducing sudden mortality. Adaptogenic potential is an umbrella term for resilience to stress, immunity and productivity of the animals These biological components show an augmented effect to prolong the lifespan of an animal. There is a need to study the mode of action of the phytogenics for their adaptogenic potential.

Phytocee^™, is a synergistic combination of these globally known adaptogens. It has been shown to have a significant impact on thermogenic stress markers. Pharmacological validations to study the mechanism of action of Phytocee^™ (200g/ton feed) has shown an influence on thermogenic stress. [5–6] As scientific research progresses, the in-silico tools reveal the possibility of deducing and identifying evidence-based mechanisms of actions involving bioenergetics, anti-inflammatory, and antioxidative mechanisms that were decoded. These mechanisms were strongly associated with enhancing the adaptogenic potential, contributing to the prolonged lifespan of the animals. For this purpose, various in-silico models were used to predict the possible pharmacological mechanism of action of the phytochemicals in increasing the longevity of the animals.

The signaling regulates longevity mechanisms; the binding interactions of insulin and its receptor activate the P13K-Akt signaling pathway to produce AC-mediated cAMP, acting on the CREB and further activating the FOXO signaling to employ cellular free radical scavengers SOD2 and CAT that are involved in cell detoxification, the mTOR signaling pathway for inhibition of redundant protein synthesis. The literature has highlighted the importance of insulin signaling, mTOR, and other proteins and pathways in regulating a cell's longevity. [7] Studies in C. elegans have shown that autophagy genes are essential for the lifespan extension observed in mutants with reduced IIS, mTOR signaling, or dietary restriction. This suggests that autophagy acts downstream of these pathways to mediate their effects on longevity. [8] Endurance plays a significant role in regulating longevity at the cellular level by signaling through the AMPK pathway, which is activated upon sensing low cellular energy levels and acting on sirtuin to promote mitochondrial biogenesis and maintain cellular bioenergetics. Literature studies have reported that the overexpression of SIRT6 enhances DNA repair mechanisms and maintains telomere integrity, contributing to improved cellular function during aging [9].

Thus, the present study illustrates the role of phytogenics through the validated presence of unique phytochemicals, as determined by LC-MS/MS, and unveils their role in adaptogenic activity via longevity mechanisms, as elucidated through in-silico studies.

Materials and Methods

LC–MS Conditions

The molecular ion profiles and fragmentation patterns of the selected phytoconstituents were investigated using a Sciex QTRAP 4500 triple-quadrupole Mass spectrometer, coupled with a Shimadzu LC-40D XR, in both positive and negative electrospray ionization modes, within the scan range of m/z 50–550, using Analyst software to record the data.

Target identification and retrieval of protein sequences

The target proteins were identified through a systematic literature review and by searching curated pathway databases implicated in pathways relevant to linking stress, insulin resistance, mitochondrial biogenesis, and protein processing accuracy, which regulate cellular longevity. The target genes, along with their annotations, were retrieved in FASTA format through UniProtKB https://www.uniprot.org/. Forkhead box O (FOXO1, FOXO3), peroxisome proliferator-activated gamma receptor coactivator alpha (PPARGCA), sirtuin 1 (SIRT1), mechanistic target of rapamycin (mTOR), adenosine monophosphate kinase (AMPK), and other pathways relevant to enhancing cell longevity were subjected to ShinyGO0.82 https://bioinformatics.sdstate.edu/go/.

Gene enrichment & KEGG pathway

The ligand files retrieved from https://pubchem.ncbi.nlm.nih.gov/ were uploaded to the DIGEP Pred2.0 https://www.way2drug.com/digep-pred/ for the regulation of unique target genes. The entire protocol was followed at the activator Pa > 0.5. The modulated gene set was mapped on an enrichment bubble in INPUT2.0 https://cbcb.cdutcm.edu.cn/INPUT/Home/ based on the gene count and p values. Gene network analysis was done for the data generated. The genes linking to the longevity regulation pathways were analyzed for the following parameters: gene count, P-value, signal and strength. Gene count indicates the number of relevant genes that were modulated. P-value indicates the significance of modulated genes in the gene set. Some genes may have high significance, but they may be less in count, and some genes may be high in count and have low p values or of less significance. Signal gives a balance between the two metrics. Strength parameter indicates a higher count of genes with more significance. The modulated gene set created by the DIGEP Pred2.0 tool https://www.way2drug.com/digep-pred/ was mapped on the KEGG genome database to view the longevity-regulating pathway analysis (KEGG ID: hsa04211). The annotation of the signaling pathway was extracted from the (NCBI). The reactome pathways diagrams for the differentially expressed genes were produced to elucidate the cellular signaling pathways using https://reactome.org/.

Venn diagram and Heatmap analysis

A Venn diagram was drawn based on the modulated gene set. The modulated gene sets were mapped on a matrix heatmap in the SR-PLOT https://www.bioinformatics.com.cn/en based on: Phytochemicals present in the Phytocee as gallic acid (1), ellagic acid (2), ursolic acid (3), withaferin A (4).

Results and Discussion

Monogastric animals reared globally are exposed to various internal and environmental stressors; therefore, enhancing their resistance to stress is a pressing need for the continued exponential growth of the economy. Stress is inversely proportional to the lifespan of the animals. Therefore, reducing stress and enhancing cell longevity is of utmost importance. This can be achieved by strengthening the adaptogenic potential of the animals, which enables them to overcome the significant stress imposed by environmental conditions. This product consisted of adaptogenic classes of hydrolysable tannins, triterpenoids, and withanolides whose structures were analyzed for chemical composition in LC-MS/MS. The phytochemicals from Phytocee™ were confirmed by LC-MS/MS from different classes with structures indicated in Fig. 1. Triterpenoids, namely ursolic acid, exhibited a protonated molecular ion at m/z 457 [M + H] +. The MS/MS fragmentation produced a major ion at m/z 439 [M + H − H2O] +, indicating a neutral loss of water. Further fragmentation yielded ions at m/z 411 [M + H − COOH] + and m/z 393, confirming the triterpenoid skeleton of ursolic acid. The withanolides, such as withaferin A, exhibited a protonated molecular ion at m/z 471 [M + H] +, which upon fragmentation yielded a major ion at m/z 453 [M + H − H2O] +, indicating the elimination of water. Additional ions at m/z 435 [M + H − 2H2O] + and m/z 417 confirmed successive water losses, characteristic of withanolide lactone fragmentation. The hydrolysable ion gallic acid was detected in the negative ion mode with a molecular ion at m/z 169 [M − H] −. The MS/MS spectrum exhibited fragment ions at m/z 125 [M − H− CO2] −, corresponding to the decarboxylation of gallic acid, and m/z 97 [M − H − COOH − CO] −, confirming its polyphenolic structure. The hydrolyzable tannins, namely ellagic acid, showed a deprotonated molecular ion at m/z 301 [M − H] −, with characteristic fragmentation ions at m/z 257 [M-H-CO2] − and m/z 229 [M-H − COOH − CO] −, corresponding to sequential decarboxylation and carbonyl losses. The other tannins were confirmed in negative ionization mode as gallic acid (m/z 169) fragmented to yield m/z 125 and 97, indicating decarboxylation and further loss of CO groups, while ellagic acid (m/z 301) produced fragment ions at m/z 257 and 229, corresponding to successive CO₂ and COOH eliminations. Collectively, these fragmentation patterns agree with literature reports, thereby confirming the structural identity and authenticity of the compounds analyzed, which can be further taken up for in-silico studies. The analytical results of the compounds are shown in Supplementary Figure S1.

Fig. 1

Phytochemicals present in the Phytocee as gallic acid (1), ellagic acid (2), ursolic acid (3), withaferin A (4)

The gene datasets that were collected from DIGEP Pred 2.0 (https://www.way2drug.com/digep-pred/) and the results were recorded in Table 1. The parameters: gene counts, p-values, signal, and strength for all the active constituents were recorded in Table 2.

Table 1
Modulated gene list for longevity regulating pathway (KEGG ID: hsa04211).
KEGG Pathways	Phytocee	Modulated gene list
	Withaferin A	ADCY3, ADCY5, AKT2, AKT3, ATF2, ATF4, BAX, CREB3L2, FOXO1, FOXO3, IRS2, NFKB1, NRAS, PIK3CA, PIK3R1, PPARG, PRKAB1, PRKAG2, SESN2, SESN3, SIRT1, SOD2, TP53, TSC1, ULK1
	Gallic acid	ADCY3, ADCY5, AKT2, AKT3, ATF4, ATG101, ATG5, BAX, CREB3, CREB3L1, CREB3L2, EIF4E, FOXO1, FOXO3, IGF1R, IRS2, KL, KRAS, NFKB1, NRAS, PIK3CA, PRKAB1, PRKAG2, RELA, SIRT1, TP53, TSC1, ULK1
Longevity regulating Pathway	Ursolic acid	ADCY3, ADCY5, AKT2, AKT3, ATF4, ATG101, ATG5, BAX, CREB3, CREB3L1, CREB3L2, EIF4E, FOXO1, FOXO3, IGF1R, IRS2, KL, KRAS, NFKB1, NRAS, PIK3CA, PRKAB1, PRKAG2, RELA, SIRT1, TP53, TSC1, ULK1
	Ellagic acid	ADCY3, ADCY4, ADCY5, ADCY7, ADCY9, ADIPOQ, ADIPOR1, AKT1, AKT1S1, AKT2, ATF2, ATF4, ATG101, ATG5, BAX, CAT, CREB1, CREB3, CREB3L1, CREB3L2, CREB5, EIF4E, EIF4EBP1, FOXO3, HRAS, IGF1, IGF1R, INS, IRS1, IRS2, KL, KRAS, MTOR, NFKB1, PIK3CA, PIK3CB, PIK3CD, PIK3R1, PIK3R2, PIK3R3, PPARG, PPARGC1A, PRKAB1, PRKACA, PRKAG2, RB1CC1, RELA RPTOR, SESN1, SESN2, SIRT1, SOD2, TP53, TSC1, TSC2, ULK1, ADCY1, ADCY2, ADCY3, ADCY6, ADIPOR2, AKT3, APPL1, ATF6B, CAMK4, CAMKK2, CREB3L3, EHMT1, IRS4, NRAS, PRKAA2 PRKAB2, PRKACB, PRKAG1, RPS6KB1, SESN3

Table 2
Modulated genes with different parameters to measure their significance
Longevity regulating pathway
Phytocee® Compounds	False discovery rate	No. of modulated genes	Signal
Ellagic acid *	1.27e-159	74 out of 87	30.94
Gallic acid *	3.27e-130	60 out of 87	26.88
Withaferin A	5.1e-50	25 of 87	15.94
Ursolic acid	1.24e-58	27 of 87	16.18

* The values denote highly significant modulation of genes regulating the longevity pathways (p < 0.01)

All the Phytocee^™ compounds significantly modulated a total of 87 genes. Out of all the compounds, withaferin A showed the highest degree of modulation (FDR: 1.27e-159). It significantly modulated 74 genes, including genes responsible for stress and insulin resistance signaling, such as FOXO1, FOXO3, SIRT1, and CREB3L2. In the order of decreasing degree of modulation, gallic acid showed significant modulation (FDR: 3.27 e-130). A total of 60 genes involved in cellular energy homeostasis, stress, and insulin resistance signaling pathways, such as FOXO1, FOXO3, IGF1, and PPARGC1A, were modulated with great significance by gallic acid. Highly significant modulation was shown by both ellagic acid and ursolic acid on the longevity-regulating genes. The genes that were modulated significantly for the pathways that help in promoting cellular fitness and longevity. The significant cellular pathways studied for the differentially expressed genes are demonstrated in Fig. 2.

Fig. 2

Significant gene pathway network enrichment mapping, KEGG and Reactome for longevity regulation pathways using STRING2.0, INPUT2.0, ShinyGO0.82 and Reactome software.

High levels of significance were observed for MAPK, GPCR, and mTOR signaling pathways, suggesting energy biogenesis, protein synthesis, and modification of a living cell. The biological process of oxidative scavenging with the help of macroautophagy was also modulated by the compounds with great significance. A wide range of research articles have been published on the anti-stress, adaptogenic and pro-longevity properties of these phytochemicals. Withaferin A has been shown to activate AMP-activated protein kinase (AMPK) in myocardial tissue of mice thereby having an action on infarcts in mice [10]. Many clinical and animal trials have proved the efficacy of Withaferin A alone to show a significant reduction in stress levels which is attributed to downregulation of HPA axis activity and the root extracts showing lowered cortisol production and epinephrine levels post stress inducing exercises which added to a support to adaptogenic properties of a compound [11–13]. Researchers have proved that ellagic acid has the potential to regulate AMPK/P13k/mTOR pathways, thereby significantly reducing the angiogenic signaling [14]. Some experimental gene expression studies showed that supplementation of ellagic acid displayed upregulated expressions of CAT, dFOXO, ATG1, and SOD2 in ellagic acid-treated male flies, and upregulated expressions of dFOXO, CAT, and SOD2 in ellagic acid-treated female flies, which indicated the pro-longevity effects of the ellagic acid [15]. In the present study, the biological significance was also observed in the reactome pathways. In a few studies, Ursolic acid alone has been able to show adipogenesis-inhibiting effects by activation of the AMPK pathway. It plays a significant role in muscle protein anabolism through the mTOR pathway [16–17]. The direct impact of gallic acid alone was observed in the findings of Zhang et.al. (2023) on lipid metabolism via the AMPK-ACC-PPARα axis [18]. Gallic acid has also been found to show active inhibition of the PI3K-Akt signaling pathway [19].

Fig. 3

A schematic Venn diagram and heat map representing the significantly modulated pathways linking to regulating longevity.

The synergistic action of the compounds present in Phytocee ^TM was reported by constructing the flower plot Venn diagram and heat maps to study the significance of the differentially expressed genes linking to the cellular longevity. Figure 3. Highly significant modulation (34 genes) was shown by the synergistic action of gallic acid and ellagic acid. It also represented all the common modulated genes (17 genes) by the product Phytocee^™. The matrix heatmap showed that the phytochemicals indicated excellent modulation in the pathways determining the resistance and endurance of a cell exposed to significant amounts of stress. This synergy has been proven by Gallic acid and Withaferin A to modulate the AMPK pathway, improving energy efficiency and autophagy [20]. It enhances endurance, improves immune response, regulates stress hormones, and increases the longevity of cells, thereby reducing mortality in animals and contributing significantly to market potential.

Conclusion

The current elucidative mechanistic approaches have played a major role in deciphering the longevity regulating pathways. This may be a strong indicator of reducing mortality amongst the animals associated with significant economic concerns. The present in-silico data has helped to draw predictive conclusions on the synergistic activity of the Phytocee^™ polyherbal combination upon the energy sensing, stress responses, insulin signaling, and post-translational modifications. This research has broadened our horizon in the areas of adaptogenic activity of natural phytochemicals and their role in regulating the longevity of a cell at molecular level. This study provides an excellent framework for developing a mechanism of action using cutting-edge technology, laying the groundwork for future studies.

Electronic Supplementary Material

Below is the link to the electronic supplementary material

Supplementary Material 1

Author Contribution

Shamana K and Edwin R contributed in curation and the analysis of the original database. Jaishree SP has supported in generating the analytical data and phytochemical figures. Prashanth D and Aboli G contributed in the supervision of the data. Deepak M contributed in reviewing of the draft manuscript and supplementary files and providing resources required to publish this data.

References

Sprengel M, Laskowski R, Jost Z (2025) Withania somnifera (Ashwagandha) supplementation: a review of its mechanisms, health benefits, and role in sports performance. Nutr Metab (Lond) 22:9. https://doi.org/10.1186/s12986-025-00902-7

Gupta A, Gupta P, Bajpai G Tinospora cordifolia (Giloy): An insight on the multifarious pharmacological paradigms of a most promising medicinal ayurvedic herb, Heliyon, 10, Issue 4, 2024,e26125, ISSN 2405–8440, https://doi.org/10.1016/j.heliyon.2024.e26125

ościniak A, Stasiłowicz-Krzemień A, Szeląg M, Pawlak J, Skiera I, Kwiatkowska H, Nowak N, Bernady K, Trzaskoma P, Zimak-Krótkopad O, Cielecka-Piontek J (2025) Bacopa monnieri: Preclinical and Clinical Evidence of Neuroactive Effects, Safety of Use and the Search for Improved Bioavailability. Nutrients 17(11):1939. 10.3390/nu17111939PMID: 40507208; PMCID: PMC12158153

Wahab S, Annadurai S, Abullais SS, Das G, Ahmad W, Ahmad MF, Kandasamy G, Vasudevan R, Ali MS, Amir M (2021) Glycyrrhiza glabra (Licorice): A Comprehensive Review on Its Phytochemistry, Biological Activities, Clinical Evidence and Toxicology. Plants (Basel) 10(12):2751. 10.3390/plants10122751PMID: 34961221; PMCID: PMC8703329

Selvam R, Suresh S, Saravanakumar M, Chandrasekaran CV, Prashanth D (2018) Alleviation of Heat Stress by a Polyherbal Formulation, Phytocee™: Impact on Zootechnical Parameters, Cloacal Temperature, and Stress Markers. Pharmacognosy Res 10(1):1–8. https://doi.org/10.4103/pr.pr_138_17

Wiono AA, Wijayanti MEN, Primacitra DY, Purnawarman T, Andriyanto, Mustika AA, Fadholly A (2023) Impact of PhytoceeTM supplementation on heat stress condition in broiler chicken. Adv Anim Vet Sci 11(7):1152–1158

Pan H, Finkel T (2017) Key proteins and pathways that regulate lifespan. J Biol Chem 292(16):6452–6460. https://doi.org/10.1074/jbc.R116.771915

Tóth ML, Sigmond T, Borsos E, Barna J, Erdélyi P, Takács-Vellai K, Orosz L, Kovács AL, Csikós G, Sass M, Vellai T (2008) Longevity pathways converge on autophagy genes to regulate life span in Caenorhabditis elegans. Autophagy 4(3):330–338. https://doi.org/10.4161/auto.5618

Dolivo D, Hernandez S, and Tanja Dominko (2016). Cellular lifespansenescence: a complex balance between multiple cellular pathways. BioEssays 38 : S33-S44

10.

Yuan SY, Liu J, Zhou J, Lu W, Zhou HY, Long LH, Hu ZL, Ni L, Wang Y, Chen JG, Wang F (2016) AMPK Mediates Glucocorticoids Stress-Induced Downregulation of the Glucocorticoid Receptor in Cultured Rat Prefrontal Cortical Astrocytes. PLoS ONE 11(8):e0159513. https://doi.org/10.1371/journal.pone.0159513

11.

Andres-Mateos E, Brinkmeier H, Burks TN, Mejias R, Files DC, Steinberger M, Soleimani A, Marx R, Simmers JL, Lin B, Finanger Hedderick E, Marr TG, Lin BM, Hourdé C, Leinwand LA, Kuhl D, Föller M, Vogelsang S, Hernandez-Diaz I, Vaughan DK, de la Alvarez D, Lang F, Cohn RD (2013) Activation of serum/glucocorticoid-induced kinase 1 (SGK1) is important to maintain skeletal muscle homeostasis and prevent atrophy. EMBO Mol Med 5(1):80–91. 10.1002/emmm.201201443Epub 2012 Nov 19. PMID: 23161797; PMCID: PMC3569655

12.

Guo R, Gan L, Lau WB, Yan Z, Xie D, Gao E, Christopher TA, Lopez BL, Ma X, Wang Y (2019) Withaferin A Prevents Myocardial Ischemia/Reperfusion Injury by Upregulating AMP-Activated Protein Kinase-Dependent B-Cell Lymphoma2 Signaling. Circulation journal: official J Japanese Circulation Soc 83(8):1726–1736. https://doi.org/10.1253/circj.CJ-18-1391

13.

Della Porta M, Maier JA, Cazzola R Effects of Withania somnifera on Cortisol Levels in Stressed Human Subjects: A Systematic Review. Nutrients 2023, 15, 5015. https://doi.org/10.3390/nu15245015

14.

Mikulska P, Malinowska M, Ignacyk M, Szustowski P, Nowak J, Pesta K, Szeląg M, Szklanny D, Judasz E, Kaczmarek G, Ejiohuo OP, Paczkowska-Walendowska M, Gościniak A, Cielecka-Piontek J (2023) Ashwagandha (Withania somnifera)-Current Research on the Health-Promoting Activities: A Narrative Review. Pharmaceutics 15(4):1057. 10.3390/pharmaceutics15041057PMID: 37111543; PMCID: PMC10147008

15.

Grogan PT, Sleder KD, Samadi AK et al (2013) Cytotoxicity of withaferin A in glioblastomas involves induction of an oxidative stress-mediated heat shock response while altering Akt/mTOR and MAPK signaling pathways. Invest New Drugs 31(3):545–557 PubMed

16.

Wiciński M, Fajkiel-Madajczyk A, Kurant Z, Kurant D, Gryczka K, Falkowski M, Wiśniewska M, Słupski M, Ohla J, Zabrzyński J (2023) Can Ashwagandha Benefit the Endocrine System? - A Review. Int J Mol Sci 24(22):16513. 10.3390/ijms242216513PMID: 38003702; PMCID: PMC10671406

17.

Kowshik J, Giri H, Kishore TK, Kesavan R, Vankudavath RN, Reddy GB, Dixit M, Nagini S (2014) Ellagic acid inhibits VEGF/VEGFR2, PI3K/Akt and MAPK signaling cascades in the hamster cheek pouch carcinogenesis model. Anti-cancer Agents Med Chem 14(9):1249–1260. https://doi.org/10.2174/1871520614666140723114217

18.

He Y, Li Y, Zhao T, Wang Y, Sun C (2013) Ursolic acid inhibits adipogenesis in 3T3-L1 adipocytes through LKB1/AMPK pathway. PLoS ONE 8(7):e70135. 10.1371/journal.pone.0070135PMID: 23922935; PMCID: PMC3724828

19.

Ogasawara R, Sato K, Higashida K, Nakazato K, Fujita S (2013) Ursolic acid stimulates mTORC1 signaling after resistance exercise in rat skeletal muscle. Am J Physiol Endocrinol Metab 305(6):E760–E765. https://doi.org/10.1152/ajpendo.00302.2013

20.

Zhang J, Zhang W, Yang L, Zhao W, Liu Z, Wang E, Wang J (2023) Phytochemical gallic acid alleviates nonalcoholic fatty liver disease via AMPK-ACC-PPARa axis through dual regulation of lipid metabolism and mitochondrial function. Phytomedicine: Int J phytotherapy phytopharmacology 109:154589. https://doi.org/10.1016/j.phymed.2022.154589

21.

Ho HH, Chang CS, Ho WC et al (2010) Anti-metastasis effects of gallic acid on gastric cancer cells involve inhibition of NF-κB activity and downregulation of PI3K/Akt/small GTPase signals. Food Chem Toxicol 48(8–9):2508–2516 Frontiers

22.

Tang Y, Zhang X, Wang Y et al (2022) Dietary ellagic acid ameliorated Clostridium perfringens-induced subclinical necrotic enteritis in broilers via regulating inflammation and cecal microbiota. J Anim Sci Biotechnol 13:47. https://doi.org/10.1186/s40104-022-00694-3

23.

Li ZP, Henning SM, Lee RP, Lu QY, Summanen PH, Thames G et al (2015) Pomegranate extract induces ellagitannin metabolite formation and changes stool microbiota in healthy volunteers. Food Funct 6(8):248795. https://doi.org/10.1039/c5fo00669d

24.

Vinodkumar S, Santhanu K, Natarajan K, Senthil K (2021) An in silico approach towards exploration of the oxidative stress resistance of major withanolides of Withania somnifera in relation to COVID-19 management. J Phytology 13:192–202. https://doi.org/10.25081/jp.2021.v13.7356

25.

Hebbar S, Nandan SK, Shetty CR, Nayak P, Dhas N, Naha A (2022) In-silico screening and molecular docking studies of active constituents of Withania somnifera to investigate its kinase inhibitory activities. J Pharm Negat Results 13(4):1342–1349. https://doi.org/10.47750/pnr.2022.13.04.187

26.

Xiao Y, Huang R, Wang N, Deng Y, Tan B, Yin Y, Qi M, Wang J (2022) Ellagic Acid Alleviates Oxidative Stress by Mediating Nrf2 Signaling Pathways and Protects against Paraquat-Induced Intestinal Injury in Piglets. Antioxid (Basel Switzerland) 11(2):252. https://doi.org/10.3390/antiox11020252

27.

Kharat P, Sarkar P, Mouliganesh S, Tiwary V, Priya VBR, Sree NY, Annapoorna HV, Saikia DK, Mahanta K, Thirumurugan K (2020) Ellagic acid prolongs the lifespan of Drosophila melanogaster. GeroScience 42(1):271–285. https://doi.org/10.1007/s11357-019-00135-6

28.

Yang X, Pan X, Jia Z, Bai B, Zhi W, Chen H, Ma C, Ma D (2022) Oral administration of Lactobacillus brevis 23017 combined with ellagic acid attenuates intestinal inflammatory injury caused by Eimeria infection by activating the Nrf2/HO-1 antioxidant pathway. Vet Res 53:21. 10.1186/s13567-022-01042-z

29.

Naghibi N, Sadeghi A, Movahedinia S, Rahimi Naiini M, Rajizadeh MA, Bahri F, Nazari-Robati M (2023) Ellagic acid ameliorates aging-induced renal oxidative damage through upregulating SIRT1 and NRF2. BMC Complement Med Ther 23:77. 10.1186/s12906-023-03907-y

30.

Tanaka M, Kishimoto Y, Sasaki M, Sato A, Kamiya T, Kondo K, Iida K (2018) Terminalia bellirica (Gaertn.) Roxb. Extract and Gallic Acid Attenuate LPS-Induced Inflammation and Oxidative Stress via MAPK/NF-κB and Akt/AMPK/Nrf2 Pathways. Oxidative Med. Cell. Longev. ; 2018:9364364. 10.1155/2018/9364364

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Unveiling novel longevity-regulating mechanisms of phytogenic adaptogen Phytocee TM through a computational biology approach

Abstract

Phytogenics is a promising and novel sector in animal health. Indian botanicals have been shown to protect animals from various diseases and health stressors, which include nutritional, environmental, biological, and chemical stressors. These stressors may cause sudden mortality in the animals and are a matter of concern among farmers, which PhytoceeTM can address. It consists of scientifically validated (LC-MS/MS) phytochemicals belonging to the classes of triterpenoids, hydrolysable tannins, and withanolides that help enhance the adaptogenic potential of the animals. The adaptogenic potential of an animal is associated with prolonging their lifespan. This research investigated the pathway-focused approaches for increasing longevity in animals at a cellular level by observing their differentially expressed genes (DEGs): FOXO, AMPK, SIRT1, and mTOR using computational tools for gene enrichment: DIGEP Pred2.0, which showed significant modulation of the DEGs (1.25e-159), especially for the enriched term “longevity regulating pathway”. Pathway mapping using SHINYGO0.82, KEGG, showed significant mapping of these genes in the enriched pathway. Network pharmacology was done to detect protein-protein interactions for these DEGs using INPUT2.0. The heat maps and flowerpot Venn diagrams were generated for enriched pathways using SR-PLOT, which showed that the maximum genes were responsible for regulating the longevity of the animals. The study concluded with a possible molecular pharmacology of PhytoceeTM, suggesting that the product may have helped overcome stressors by increasing the lifespan of various animal species, including poultry, swine, and aquatic animals. The possibility can be further validated through the wet lab trials.