Search Results (58)

Background/Objectives: Hyperuricemia (HUA) is a metabolic disease with increasing incidence. Chicory (Cichorium intybus L.), a traditional medicinal and edible plant, has demonstrated anti-HUA effects. However, the metabolic profiles of its aerial parts and roots are still not fully characterized. Moreover, few studies have investigated its anti-HUA effects using cell metabolomics. Methods: The metabolomes of chicory root and aerial parts were characterized using UPLC-QTOF-MS-based untargeted metabolomics. Subsequently, the anti-HUA mechanism of chicory root was investigated by performing non-targeted metabolomics in HK-2 cells. Results: The results demonstrated that various hydroxycinnamic acids and flavonoids were more abundant in aerial parts, whereas sesquiterpenes and oligosaccharides were characteristic of the root. Both chicory root and aerial part extracts significantly reduced uric acid (UA) levels in HK-2 cells induced by adenosine with xanthine oxidase (XO). Cellular metabolomic profiling indicated a distinct separation between the root extract (CR40, 40 mg/mL) and the model group. OPLS-DA identified 165 differential metabolites, including acylcarnitines, acylamino acids, peptides, phospholipids, glycerides, and lipid-like molecules. These metabolites were associated with key metabolic pathways of sphingolipids, glycerophospholipids, phosphonate and phosphinate, linoleic acid, biotin, purine, as well as taurine and hypotaurine metabolism. Conclusions: Chicory is rich in diverse bioactive compounds and exhibits significant anti-HUA activity by modulating multiple metabolic pathways. Full article

Renal urate deposition is a pathological inflammatory condition characterized by the accumulation of urate crystals in the kidneys, resulting from uric acid supersaturation. Cichorium intybus L. (chicory) is a traditional medicinal herb recognized for its efficacy in treating hyperuricemia and gout; however, its effectiveness and underlying mechanisms in mitigating renal urate deposition remain inadequately understood. This study investigates the role of the ATP-binding cassette sub-family G member 2 (ABCG2) transporter and the lncRNA H19/miR-21-3p in renal urate deposition, while also validating the therapeutic effects and mechanisms of chicory extract. Renal urate deposition was induced in rats through the administration of potassium oxonate, adenine, yeast extract, and lipopolysaccharide. The levels of serum uric acid (SUA), urate deposition, inflammation, renal function, and histological changes were analyzed. Dual-luciferase assays, reverse transcription quantitative polymerase chain reaction (RT-qPCR), and immunohistochemistry were utilized to elucidate the relationship among ABCG2, lncRNA H19, and miR-21-3p. The chemical composition and active ingredients of chicory were analyzed using UPLC-LTQ-Orbitrap-MS, along with molecular docking and cell experiments. In rats with renal urate deposition, serum UA levels were elevated, renal UA excretion was reduced, and levels of low inflammatory factors, such as interleukin-6 (IL-6), tumor necrosis factor-alpha (TNF-α), and hypersensitivity C-reactive protein (hs-CRP), were increased. Additionally, significant renal tissue damage accompanied the urate deposition. Notably, these abnormalities were substantially reversed following treatment with chicory extract. A dual-luciferase reporter assay confirmed the regulatory relationships among miR-21-3p, lncRNA H19, and ABCG2. Immunohistochemical analysis and RT-qPCR demonstrated a significant upregulation of miR-21-3p expression, alongside a downregulation of lncRNA H19, ABCG2 mRNA, and ABCG2 expression in the kidney tissue of rats with renal urate deposition. Chicory extract may exert its inhibitory effect on renal urate deposition by regulating the lncRNA H19/miR-21-3p axis to enhance ABCG2 expression. Furthermore, UPLC-LTQ-Orbitrap-MS identified 69 components in the chicory extract, including scopoletin, quercetin-3-O-β-D-glucuronide, 11β,13-dihydrolactucopicrin, and kaempferol-3-O-β-D-glucuronide, which were absorbed into the blood of both normal rats and those with renal urate deposition. Molecular docking and cell experiment further validated the effective regulation of 11β,13-dihydrolactucopicrin in ABCG2 and the lncRNA H19/miR-21-3p axis. The downregulation of ABCG2, mediated by the lncRNA H19/miR-21-3p axis, may represent a critical pathogenic mechanism in renal urate deposition. Chicory alleviates this deposition by modulating the lncRNA H19/miR-21-3p axis to enhance ABCG2 expression, potentially through its component, 11β,13-dihydrolactucopicrin, thereby revealing novel therapeutic insights for renal urate deposition. Full article

Hyperuricemia (HUA) is a metabolic disorder characterized by abnormal purine metabolism and/or reduced uric acid (UA) excretion. Chicory (Cichorium intybus L.), recognized in Traditional Chinese Medicine, is noted for its anti-HUA effects, particularly in enhancing intestinal UA excretion, though the underlying mechanisms remain unclear. Studies indicate that disruptions in gut microbiota and its metabolites are associated with HUA, and chicory has been demonstrated to ameliorate gut microbiota dysbiosis. Among gut microbiota-derived metabolites, butyrate, a short-chain fatty acid, plays a crucial role in gut functions and is linked to HUA. Therefore, butyrate may be pivotal in elucidating the mechanism by which chicory promotes intestinal UA excretion. This study aims to investigate whether chicory facilitates intestinal UA excretion through gut microbiota-derived butyrate and to elucidate the underlying mechanism. We employed an integrated methodology combining network biology with the NHANES database analysis to explore the pathological relationship between butyrate and HUA. Our findings were subsequently validated through animal experiments. We administered chicory to rats with HUA to ascertain whether butyrate serves as the key gut microbiota metabolite through which chicory promotes intestinal UA excretion. Furthermore, we utilized western blotting to assess the expression of core targets within the PPARγ-ABCG2 pathway associated with butyrate under conditions where animals received butyrate supplements and PPARγ agonists separately. The network biology indicates that butyrate is a crucial short-chain fatty acid influencing HUA. Analyses of NHANES data and animal experiments further confirm a significant negative correlation between butyrate and serum uric acid (SUA) levels. HUA rats exhibited intestinal barrier damage, impaired intestinal UA excretion, reduced butyrate levels, and decreased expression of PPARγ and ABCG2 proteins. Intervention with chicory in HUA rats repaired intestinal barrier damage, enhanced intestinal UA excretion, and increased both butyrate levels and the expression of PPARγ and ABCG2 proteins. Similarly, interventions with butyrate supplements or PPARγ agonists in HUA rats effectively promoted intestinal UA excretion and increased the expression of PPARγ and ABCG2 proteins. This study demonstrates that butyrate is a key metabolite produced by gut microbiota, through which chicory regulates gut microbiota to enhance intestinal UA excretion. The underlying mechanism involves the activation of the PPARγ-ABCG2 pathway, which is facilitated by elevated butyrate levels in the intestine. Full article

Cichorium intybus L. (common chicory) is a medicinal plant valued for health-promoting effects. Although analgesic properties are known for chicory sesquiterpenes, the effects of extracts need yet to be explored. This study aimed to evaluate for the first time the analgesic effect (against nociceptive pain) of the root extract from C. intybus var. foliosum. The target evaluation was preceded by toxicity tests in vivo and phytochemical standardization of root extracts prepared with different extraction methods—pectinase-assisted, pressure-assisted, and a combination of both—to choose the most effective one. The phytochemical profiling involved UHPLC-PDA-ESI-MS/MS and UHPLC-PDA analyses. The toxicity and the analgesic effects were tested in mice following the OECD 423 guideline and the hot-plate test, respectively. The highest recovery of bioactive compounds was achieved for the pressure-assisted extract: 642.5 mg sesquiterpene lactones, 187.1 mg phenolic acids, and 47.3 g inulin/100 g of dry matter. The extract showed no toxic effects at the oral dose of 2000 mg/kg body weight, including no histopathologic changes, in mice within two weeks (GHS Category 5/Uncategorized). The maximum analgesic effect (MAE) of the extract at 600 mg/kg was 6.75% for rearing and 13.7% for jumping, with the impact on the nocifensive reactions not differing significantly from those of paracetamol at 60 mg/kg. Despite the relatively low effects at 600 mg/kg, the verified safety and abundance of active compounds encourage further studies on the extract and its active fractions as potential approaches to complementary pain therapy, with special concern for their mechanisms of action. Full article

Chicory (Cichorium intybus L.) roots are valued in medicine for their potential health benefits. Producing callus from chicory roots through tissue culture technology can streamline bioactive metabolites production and ensure a sustainable supply chain. The current study explored the impact of plant growth regulators (PGRs) and light conditions on the characteristics of callus induced from C. intybus root explants. The effect of fungal elicitors [yeast extract (YE), Fusarium oxysporum, and Aspergillus niger] on bioactive metabolite production from root-derived callus was investigated. Callus color varied notably between a 16/8 h light/dark cycle and complete dark, with differences in texture based on PGR concentrations and light conditions. High weights of callus formed were generally recorded under the 16/8 h light/dark cycle. Low concentrations of YE (1 g/L) and F. oxysporum (0.25 g/L) enhanced callus biomass fresh weight, while high concentrations of A. niger (1 g/L) improved callus dry matter significantly. The content and productivity of total phenolic were maximized at 1 g/L of YE and 1 g/L of F. oxysporum. Callus cultures elicited with a higher level of A. niger recorded the higher values of total flavonoid production. High-performance liquid chromatography (HPLC) analysis revealed significant variations in chlorogenic acid, catechin, and caffeic acid levels among the different elicited cultures. A. niger at 1 g/L notably increased chlorogenic acid content, while catechin levels were enhanced by specific concentrations of YE. Catalase (CAT) activity was significantly affected by different elicitors, while only the higher level of F. oxysporum and A. niger showed a significant increase in peroxidase (POD) activity. DPPH scavenging activity was elevated by all fungal elicitors. Principal Component Analysis delineated distinct variations in callus traits in response to different elicitors, with specific treatments showcasing enhanced biomass production, bioactive compound accumulation, and antioxidant activities. Through meticulous experimentation, this study paves the way for enhancing chicory root-derived products, ensuring sustainable production and potent bioactivity. Full article

Prolonged sleep deprivation impairs brain function and increases the risk of mental health disorders. Cichorium intybus L. Oligo-polysaccharides (JSO), bioactive compounds derived from chicory, belong to the category of food-medicine homologous substances, possess gut microbiota-modulating and anti-inflammatory properties, and serve as a natural prebiotic, having significant research value in food science. This research examined the anxiolytic properties of JSO in a murine model subjected to chronic sleep deprivation (CSD) stress and explored the mechanisms behind this effect, providing experimental evidence for the development of Cichorium intybus L. as a functional food. Specific pathogen-free (SPF) KM male mice were allocated at random into six experimental groups: the control group, the CSD model group, the diazepam (10 mg/kg) group, and the JSO treatment groups at low (50 mg/kg), medium (100 mg/kg), and high (200 mg/kg) doses. Following 3 weeks of CSD, anxiety-like behaviors were assessed using the open field test, elevated plus maze test, light–dark box test, forced swim test, and marble-burying test. To analyze the composition of gut microbiota, 16S rRNA sequencing was employed, while protein expression in the BDNF, PI3K/AKT/mTOR, and NLRP3 inflammasome pathways was detected by Western blot. Behavioral analysis indicated that JSO (at doses of 100 and 200 mg/kg) markedly enhanced both the time allocated to open arms and the number of entries into open arms in the elevated plus maze test (p < 0.05). JSO (at doses of 50 and 200 mg/kg) significantly elevated transitions in the light–dark box test (p < 0.05), all JSO doses drastically cut down marble-burying behavior (p < 0.001, p < 0.01, p < 0.01). The 16S rRNA sequencing indicated that JSO intervention increased Bacteroidetes abundance while reducing Actinobacteria. Western blot analysis demonstrated that JSO significantly downregulated the ratios of p-mTOR/mTOR, p-PI3K/PI3K, p-AKT/AKT, BAX/BCL-2, as well as the expression levels of NLRP3, ASC, Caspase-1, and IL-6 proteins (p < 0.05), while upregulating hippocampal BDNF (p < 0.05). These results indicate that JSO ameliorates CSD-induced anxiety-like behaviors by restoring gut microbiota homeostasis, regulating the BDNF-PI3K/AKT/mTOR and BAX/BCL-2 apoptosis pathways, and suppressing NLRP3 inflammasome-mediated neuroinflammation. Full article

Background: Cichorium intybus L., a member of the Asteraceae family, has numerous health-promoting properties that categorize its preparations as functional foods and herbal medicines. Most previous research focused on the root of C. intybus var. sativum (industrial chicory) as a rich source of inulin, while the witloof variety (C. intybus var. foliosum) is less explored. Objectives: This study aimed to evaluate the cytotoxicity of C. intybus var. foliosum root extracts obtained with different extraction protocols and to analyze their polysaccharide and polyphenol content. Methods: Freeze-dried root extracts were prepared using water and three extraction methods: pectinase-assisted, pressure-assisted, and a combination of both. The contents of inulin, total polyphenols, and total caffeic acid derivatives in the extracts were measured by the Layne–Eynon, Folin–Ciocalteu, and UHPLC-PDA methods, respectively. Cytotoxicity of the extracts and inulin was tested in vitro using the L929 cell line, MTT method, and paracetamol as the reference standard. Results: Inulin levels in the extracts ranged from 43.88 to 50.95 g/100 g dry matter (dm), total polyphenols were between 816.7 and 906.4 mg/100 g dm, and total phenolic acids ranged from 11.50 to 187.1 mg/100 dm, with pressure-assisted extraction yielding the highest phytochemical recovery. The cytotoxicity tests showed IC₅₀ values from 4.72 to 7.31 mg/mL for the extracts, compared to 3.02 for paracetamol and 19.77 for inulin. Conclusions: Given the high content of active compounds and low cytotoxicity, the root extracts of C. intybus var. foliosum merit further research into their functional and medicinal properties. Pressure-assisted extraction is recommended for effective extraction of chicory. Full article

Chicory (Cichorium intybus L., 2n = 18), belonging to the Asteraceae family, exhibits significant edible, medicinal, and pasture values. Moderate research has been performed on identifying Chicory species’ chromosomes using fluorescence in situ hybridization (FISH) and C-banding. Detailed karyotype comparisons with chromosome nomenclature have not yet been performed for Chicory and similar species. In this study, the tandem repeats (TRs) were predicted and mapped to chromosomal regions based on released C. intybus L. ASM2352571 genome assembly v1, and then compared to the genome of Lettuce (Lactuca sativa L.). Nine new oligo probes were then developed and employed for karyotypic investigation of endive, Lettuce, and Chicory mitotic metaphase using non-denaturing FISH (ND-FISH). By combining the conserved oligo probes for 5S rDNA and 18S rDNA with the unique ND-FISH signals of new TR-oligo probes, we can develop a high-resolution standard karyotype for the cultivars of Lettuce and Chicory. The occurrence of chromosome structure variations from the natural population of Chicory and Lettuce was also revealed by ND-FISH with multiple oligo probes. The current observation of the karyotype differences and divergences of Lactuca and Cichorium and the genomic research offers crucial information about the Asteraceae family’s genetic diversity, chromosomal dynamics, and evolutionary routes. Full article

The aim of this review is to discuss the numerous health-promoting properties of Cichorium intybus L. and bring together a range of publications to broaden knowledge and encourage further research and consideration of the plant use as treatment for a range of conditions. A comprehensive search of articles in Polish and English from 1986–2022 years was carried out in PubMed, Google Scholar and ScienceDirect using the keywords chicory, Cichorium intybus L., sesquiterpene lactones and their synonyms. Articles were checked for titles, abstracts, and full-text reviews. The first part of the review article discusses chicory, the countries in which it is found, its life cycle or modern cultivation methods, as well as its many uses, which will be discussed in more detail later in the article. The increased interest in plants as medicines or supplements is also briefly mentioned, as well as some limits that are associated with the medical use of plants. In the Results and Discussion section, there is a discussion of the numerous health-promoting properties of Cichorium intybus L. as a whole plant, with its collection of all the components, and we then examine the structure and the individual constituents of Cichorium intybus L. Among these, this article discusses those that can be utilized for causal applications in medicine, including sesquiterpene lactones and polyphenols, mainly known for their anti-cancer properties, although, in this article, their other health-promoting properties are also discussed. The article also examines inulin, a major component of Cichorium intybus L. The Discussion and the Conclusions sections propose directions for more detailed research and the range of factors that may affect specific results, which may have safety implications when used as supplements or medications. Full article

Background: We recently reported that extract prepared from the aerial part of Cichorium intybus L. (CE) possesses hepatoprotective, hypolipidemic, and hypoglycemic properties. This paper focuses on the effects of CE on the male rat reproductive system and the effects of this treatment on pregnancy and offspring development. Methods: The experimental male rats received 100 mg/kg bw/day, 500 mg/kg bw/day, and 1000 mg/kg bw/day of CE orally for 60 consecutive days. Rats that received tap water were used as controls. After treatment, we evaluated the effects of CE on the male reproductive system, fertility, and offspring development. Results: For CE-treated male rats, there was a significant increase in the (1) diameter of seminiferous tubules, (2) spermatogenic index, (3) number of total and motile spermatozoa, and (4) testosterone levels. Additionally, there was a decrease in the pre- and post-implantation death of the embryos in the CE-treated group. All pups born from CE-treated males demonstrated normal development. Conclusions: CE treatment significantly improved male reproductive functions. No adverse effects on pregnancy and offspring development were observed when males were treated with CE. Further clinical evaluation of CE should lead to the development of a safe and effective phytodrug for treating male infertility. Full article

The medicinal plants of the Asteraceae family are a valuable source of bioactive secondary metabolites, including polyphenols, phenolic acids, flavonoids, acetylenes, sesquiterpene lactones, triterpenes, etc. Under stressful conditions, the plants develop these secondary substances to carry out physiological tasks in plant cells. Secondary Asteraceae metabolites that are of the greatest interest to consumers are artemisinin (an anti-malarial drug from Artemisia annua L.—sweet wormwood), steviol glycosides (an intense sweetener from Stevia rebaudiana Bert.—stevia), caffeic acid derivatives (with a broad spectrum of biological activities synthesized from Echinacea purpurea (L.) Moench—echinacea and Cichorium intybus L.—chicory), helenalin and dihydrohelenalin (anti-inflammatory drug from Arnica montana L.—mountain arnica), parthenolide (“medieval aspirin” from Tanacetum parthenium (L.) Sch.Bip.—feverfew), and silymarin (liver-protective medicine from Silybum marianum (L.) Gaertn.—milk thistle). The necessity to enhance secondary metabolite synthesis has arisen due to the widespread use of these metabolites in numerous industrial sectors. Elicitation is an effective strategy to enhance the production of secondary metabolites in in vitro cultures. Suitable technological platforms for the production of phytochemicals are cell suspension, shoots, and hairy root cultures. Numerous reports describe an enhanced accumulation of desired metabolites after the application of various abiotic and biotic elicitors. Elicitors induce transcriptional changes in biosynthetic genes, leading to the metabolic reprogramming of secondary metabolism and clarifying the mechanism of the synthesis of bioactive compounds. This review summarizes biotechnological investigations concerning the biosynthesis of medicinally essential metabolites in plants of the Asteraceae family after various elicitor treatments. Full article

In traditional ryegrass/white clover (Lolium perenne L./Trifolium repens L.) pastoral systems, forage herbs such as chicory (Cichorium intybus L.) present an opportunity to fill feed deficits during late spring and summer. Although multiple research publications have evaluated the efficacy of chicory for enhancing milk production and milk fatty acid (FA) profile, no publication has quantitatively synthesised the body of research. This systematic review and meta-analysis examined the effect of chicory on milk production and composition, as well as on the milk fatty acid composition of dairy cattle. A total of 29 comparisons from 15 unique research publications involving 597 dairy cattle were used to develop a dataset for analysis. Three-level random-effect and robust variance estimator models were used to account for the hierarchical structure of the data and the dependency of effect sizes within publications. Chicory inclusion increased milk yield when compared to grass-based diets {weighted mean difference (WMD) = 1.07 (95% CI 0.54–1.60) kg/cow/d, p < 0.001}, but it provided a similar milk yield when compared to other forages such as legumes and herbs {dicots; WMD = −0.30, (95% CI −89–0.29) kg/cow/day, p = 0.312}. Increases in milk yield were congruent with differences in DM intake (p = 0.09) and ME intakes (p = 0.003), being similar in chicory-fed and dicot-fed cows but higher than grass-fed cows. Chicory feeding’s effect on milk solids was twice as high during mid lactation {154 days in milk; WMD = 0.13, (95% 0.081–0.175) kg/cow/day, p < 0.001} as during late lactation {219 days in milk; WMD = 0.06, (95% 0.003–0.13) kg/cow/day, p = 0.041}. In line with milk yield, greater and more significant effect sizes were found for alpha linolenic acid {ALA; WMD = 0.20 (95% CI 0.06–0.35) g/100 g FA, p = 0.011} when chicory was compared to grass species only. Comparing chicory with dicots suggests that chicory inclusion did not impact ALA concentrations {WMD = 0.001 (95% CI −0.02–0.2) g/100 g FA, p = 0.99}. There were no differences in conjugated linoleic acid concentration in the milk of cows fed chicory or control diets. The study provides empirical evidence of chicory’s efficacy for improved milk production and milk fatty acid composition. Full article

Using resilient, self-sustaining plants in urban green spaces enhances environmental and cultural benefits and reduces management costs. We assessed two spontaneous plant species, Linaria vulgaris Mill. and Cichorium intybus L., in four sites from the surrounding urban areas, ranging in altitude from 78 to 1040 m. Protection against UV-B radiation is crucial for plants at higher altitudes, guiding our focus on UV-visible absorption spectra, fluorometric emission spectra, secondary metabolite accumulation, and pigment dynamics in leaves. Our findings revealed a slight increase in UV-absorbing compounds with altitude and species-specific changes in visible spectra. The UV-emission of fluorochromes decreased, while red emission increased with altitude but only in chicory. Polyphenols and flavonoids showed a slight upward trend with altitude. Divergent trends were observed in condensed tannin accumulation, with L. vulgaris decreasing and C. intybus increasing with altitude. Additionally, chicory leaves from higher altitudes (792 and 1040 m) contained significantly lower triterpene concentrations. In L. vulgaris, chlorophyll pigments and carotenoids varied with sites, contrasting with UV absorbance variations. For C. intybus, pigment variation was similar to absorbance changes in the UV and VIS range, except at the highest altitude. These observations provide valuable insights into species-specific strategies for adapting to diverse environmental contexts. Full article

In Sicilian forage systems, the introduction of native self-seeding annual legumes can be beneficial in low-input farming. Intercropping would be a valuable strategy for implementing pasture resources in Sicilian forage systems during late spring. The aim of this study was to evaluate the effects on ewes’ milk production of continuous grazing with two different mixtures (i) sulla (Sulla coronarium L.), burr medic (Medicago polymorpha L.), and chicory (Cichorium intybus L.) (SuBuCh); and (ii) barrel medic (Medicago truncatula Gaertn.), snail medic (Medicago scutellata L.), and burr medic (BuSnBa). Twenty lactating ewes were homogeneously divided into four groups of five ewes. Each group was assigned to one of four 1500 m² grazing sectors consisting of two replicates of SuBuCh and BuSnBa. Ewes fed with the SuBuCh mixture showed higher milk yield, higher protein (5.17 vs. 4.85%, p < 0.001) and casein content (4.02 vs. 3.73%, p < 0.001), lower urea content (37.70 vs. 45.38 mg dL⁻¹, p < 0.001), and better clotting parameters compared to ewes grazing on the BuSnBa mixture. Finally, ewes in the SuBuCh group showed a smaller decrease in live weight at the end of the grazing period compared to BuSnBa ewes (−2.05 vs. −3.55 kg, respectively), although the difference did not reach a significant level. These preliminary one-year results seem to highlight the promising role of SuBuCh intercrop leading to a potential quantitative/qualitative improvement in grazing resources and the productive performance of grazing ewes in a semi-arid Mediterranean environment. However, it is of note that these outcomes might undergo variations when subjected to a prolonged trial extending beyond three years. Full article

The prolonged use of exogenous glucocorticoids, such as dexamethasone (Dex), is the most prevalent secondary cause of osteoporosis, known as glucocorticoid-induced osteoporosis (GIO). The current study examined the preventative and synergistic effect of aqueous chicory extract (ACE) and ethanolic purslane extract (EPE) on GIO compared with Alendronate (ALN). The phytochemical contents, elemental analysis, antioxidant scavenging activity, and ACE and EPE combination index were evaluated. Rats were randomly divided into control, ACE, EPE, and ACE/EPE MIX groups (100 mg/kg orally), Dex group (received 1.5 mg Dex/kg, Sc), and four treated groups received ACE, EPE, ACE/EPE MIX, and ALN with Dex. The bone mineral density and content, bone index, growth, turnover, and oxidative stress were measured. The molecular analysis of RANK/RANKL/OPG and Nrf2/HO-1 pathways were also evaluated. Dex causes osteoporosis by increasing oxidative stress, decreasing antioxidant markers, reducing bone growth markers (OPG and OCN), and increasing bone turnover and resorption markers (NFATc1, RANKL, ACP, ALP, IL-6, and TNF-α). In contrast, ACE, EPE, and ACE/EPE MIX showed a prophylactic effect against Dex-induced osteoporosis by modulating the measured parameters and the histopathological architecture. In conclusion, ACE/EPE MIX exerts a powerful synergistic effect against GIO by a mode of action different from ALN. Full article