Medicinal Plant Research 2024, Vol.14, No.3, 137-150 http://hortherbpublisher.com/index.php/mpr 142 Figure 2 Effects of compounds 1–3 on the expression of LPS-induced iNOS/COX-2 protein and mRNA in RAW 264.7 macrophages (Adopted from Jeong et al., 2019) Image caption: (A) Cells were activated with LPS (1 μg/mL) in presence or absence of compounds 1–3 (10 μM) for 20 h. Cell lysates were prepared and the iNOS, COX-2, and β-actin protein levels were determined by Western blotting; (B) Cells were treated with compounds 1–3 (10 μM) and/or LPS (1 μg/mL) for 6 h. The mRNA levels for iNOS, COX-2, and β-actin were determined by RT-PCR. The relative intensity of iNOS/COX-2 to β-actin bands was measured by densitometry. Veh means vehicle. The values represented mean ± S.D. of three individual experiments. * p< 0.01 indicate significant difference (* iNOS, COX-2) from LPS alone (Adopted from Jeong et al., 2019) Jeong et al. (2019) analyzed protein samples using Western blotting, showing that LPS significantly increased the protein expression of iNOS and COX-2. However, treatment with compounds 1-3 resulted in a decrease in iNOS and COX-2 protein levels (Figure 2A). This indicates that these compounds can effectively inhibit the expression of key inflammatory mediators induced by LPS during inflammatory responses. Additionally, RT-PCR was used to measure the levels of iNOS and COX-2 mRNA in cells treated with LPS and compounds 1-3. Compared to LPS treatment alone, cells treated with compounds 1-3 showed a significant decrease in iNOS and COX-2 mRNA levels (Figure 2B). These results further confirm that these compounds inhibit the expression of iNOS and COX-2 at the transcriptional level, potentially reducing inflammation mediated by these enzymes. By validating both protein and gene expression, the study clearly demonstrates the effectiveness of compounds 1-3 in inhibiting key inflammatory pathways in macrophages. This reinforces the potential of these compounds as candidates for anti-inflammatory therapy. Additionally, these compounds suppress the expression of inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2) at both protein and mRNA levels, and inhibit the nuclear translocation of nuclear factor-kappa B (NF-κB), a key regulator of inflammation (Jeong et al., 2019; Yang et al., 2020). The anti-inflammatory effects are also linked to the modulation of cytokine production, including the reduction of tumor necrosis factor-alpha (TNF-α) and interleukins (IL-1β, IL-6) (Yang et al., 2020; Qian et al., 2022). 5.2 Immunomodulatory properties A. macrocephala has been shown to possess immunomodulatory properties, which are beneficial in enhancing the immune response. Polysaccharides from A. macrocephala can activate T lymphocytes and alleviate immunosuppression induced by cyclophosphamide through the novel_mir2/CD28/AP-1 signaling pathway (Li et
RkJQdWJsaXNoZXIy MjQ4ODYzNA==