Background: Plants rich in essential oils belong to more than 50 families, including Asteraceae, Rutaceae, and Lamiaceae. These plants are widely used in food, traditional and modern medicine, perfumery, cosmetics, and pharmaceutical industries. Approximately 3,000 types of essential oils have been identified, of which more than 300 are utilized for industrial and commercial purposes. Essential oils exhibit antimicrobial, anti-inflammatory, anticancer, cytotoxic, and antioxidant activities and are therefore widely applied in medical research. In Mongolia, the extreme climate, distinct ecological regions, and the prevalence of pastoral livestock husbandry provide a scientific basis for investigating essential oil–containing plants, their chemical composition, and biological activities. Among them, Juniperus Sabina L. is of particular theoretical and practical interest. Aim: To determine the non-toxic dose of essential oil extracted from Juniper Sabina L. on neural cells. Materials and Methods: This study was conducted at the Institute of Bio-Medicine, MNUMS. Ethical approval was obtained from the Research Ethics Committee of MNUMS (Meeting date: November 21, 2025; Protocol No. 2025-26/0301). BV2 microglial and N2a neuronal cell lines were cultured under standard conditions at 37°C in a 5% CO₂ incubator. Cells were maintained in appropriate culture media, and their growth and morphology were monitored microscopically before experimentation. Cell viability was assessed using the MTT assay. BV2 microglial cells were seeded into 96-well plates at a density of 2×10⁴ cells/well and incubated for 24 hours. Essential oil was added at concentrations ranging from 50–500 µg/mL and incubated for an additional 24 hours. After treatment, MTT reagent was added and incubated for 3 hours. The resulting formazan crystals were dissolved in dimethyl sulfoxide (DMSO), and absorbance was measured at 630 nm. Cell viability was calculated based on absorbance values to determine the non-toxic concentration of the essential oil. Result: The cytotoxic effects of the essential oil were evaluated in BV2 microglial and N2a neuronal cells using the MTT assay. In N2a neuronal cells, concentrations of 50, 100, 200, and 300 µg/mL did not significantly affect cell viability, whereas 400 and 500 µg/mL reduced cell viability, indicating cytotoxic effects. Conclusion According to MTT assay results, the essential oil of Juniperus sabina showed no cytotoxic effect on N2a neuronal cells at concentrations of 50, 100, 200, and 300 µg/mL, while cytotoxic effects were observed at 400 and 500 µg/mL. In BV2 microglial cells, concentrations of 50, 100, and 200 µg/mL were non-toxic, whereas concentrations above 300 µg/mL exhibited cytotoxic effects.

Background: Dysregulation of inflammation underpins chronic cardiovascular, metabolic, neurodegenerative, and neoplastic conditions. Traditional medicine has long utilized medicinal plants to alleviate and treat systemic inflammation. Specifically, Rhodiola rosea L. and Rhodiola quadrifida (Pall.) Fisch. & C.A.Mey. possess anti-inflammatory, antitumor, antioxidant, and anti-angiogenic properties. Saposhnikovia divaricata (Turcz.) Schischk. has been shown to exert anti-inflammatory effects, support musculoskeletal function, and enhance tissue regeneration. Furthermore, Salsola laricifolia Litv. has been studied for its antioxidant, immunomodulatory, and anti-inflammatory activities. Although the individual anti-inflammatory, antioxidant, and antitumor activities of these plants are established, research evaluating the combined effect of their combined extract remains scarce, providing the rationale for this study. Aim: To evaluate the in vitro effects of a combined extract derived from these medicinal plants on the expression of selected pro-inflammatory cytokines. Materials and Methods: A dry combined extract consisting of Rhodiola rosea, Rhodiola quadrifida, Salsola laricifolia, and Saposhnikovia divaricata was prepared at the Chemistry and Technology Sector of the Institute of Pharmaceutical Research, Monos Group. An in vitro inflammation model was established using murine RAW264.7 macrophage-like cells stimulated with lipopolysaccharide (LPS, 100 ng/mL). Cell viability was assessed using the CCK-8 assay, while the output of pro-inflammatory cytokines TNF-α and IL-6 was measured via Enzyme-Linked Immunosorbent Assay (ELISA). Statistical analysis was performed using one-way ANOVA. Result: The combined plant extract demonstrated no cytotoxic effects on RAW264.7 cells across the tested concentration range. In LPS-stimulated RAW264.7 macrophages, the composite extract dose-dependently and significantly suppressed the production of the pro-inflammatory cytokines TNF-α (p<0.001) and IL-6 (p<0.0001). Conclusion 1. The combined extract of Saposhnikovia divaricata, Rhodiola rosea, Rhodiola quadrifida, and Salsola laricifolia exhibited no cytotoxicity in vitro within the range of 50–400 µg/mL. 2. The combined herbal extract significantly suppressed the production of pro-inflammatory cytokines TNF-α (p<0.01) and IL-6 (p<0.001) in LPS-induced RAW264.7 macrophages in a concentration-dependent manner.

Background: Rhododendron adamsii (Sagaan Dalya) is a medical plant widely distributed in the Altai region, Mongolia, and Russia. It is traditionally used for its calming, restorative, and immune-boosting properties. Inflammation is a complex immune response against pathogens such as bacteria, viruses, and fungi, involving macrophages, fibroblasts, mast cells, and neutrophils. These cells release inflammatory mediators such as nitric oxide (NO), TNF-α, and IL-1β. In collaboration with the Russian Foundation for Basic Research, a project was initiated to investigate the bioactive fractions of Rh. rosea (L.) and Rh. adamsii and their effects on the production of TLR4 signaling end products and associated protein activation. Previous studies within this project have shown that certain bioactive fractions exhibit anti-inflammatory activity. Specifically, fraction 7.11 suppressed NO production and inflammatory signaling molecules in LPS-stimulated RAW 264.7 macrophages, while fractions 7.46 and 7.52 influenced the phosphorylation of proteins such as ERK1/2, JNK, and IRF3. These findings suggest the need for further investigation into the effects of Rh. adamsii bioactive fractions on inflammatory signaling pathways. Aim: This study aims to evaluate the effects of bioactive fractions derived from Rhododendron adamsii on the production of TLR4 signaling end products in RAW264.7 macrophage cell cultures. Materials and Methods: The study was conducted using RAW264.7 macrophage cell cultures and bioactive fractions extracted from Rh. adamsii, dividing the experiments into three groups. After stabilization, RAW264.7 cells were stimulated with 100 ng/mL LPS. Based on previous studies, non-toxic concentrations of bioactive fractions (10 µg/mL) were applied. NO production was measured using the Griess assay, while TNF-α and IFN-β cytokine levels were evaluated using ELISA. Each experiment were repeated three times, and data were statistically analyzed using SPSS 25.0, applying One-way ANOVA and Independent Samples T-test. Results: The NO production in the positive control group was significantly higher compared to the negative control. In contrast, the experimental groups showed a statistically significant reduction in NO production, suggesting a potential inhibitory effect on TLR4 signaling in macrophages. However, fraction 7.48 reduced TNF-α levels while increasing IFN-β production, but these changes were not statistically significant. Similarly, fraction 7.55 slightly reduced TNF-α and IFN-β levels, but the effect was also statistically unsignificant. Conclusion The bioactive fractions 7.48 and 7.55 of Rh. Adamsii reduced nitric oxide (NO) production in LPS-stimulated macrophage cell line cultures, suggesting that they may inhibit TLR4 signaling. However, their lack of effect on TNF-α and IFN-β production indicates that they do not influence TLR4 signaling mediated by these cytokines. Instead, they may affect the final product output through other pathways or different stages of signal transduction.