Astragalus has antibacterial and anti-inflammatory properties. Sometimes, people use it on their skin for wound care. In addition, studies have shown that astragalus has antiviral properties and boosts the immune system, suggesting that it may help prevent colds. The extraction of dry roots or stems from plants was described in Figure 1a.
Briefly, dried roots or stems were extracted repeatedly with methanol and dichloromethane. The crude extracts (EC) were partitioned between water (H2O) and n-butanol, and the latter fraction was partitioned into 15% aqueous methanol (MeOH) and n-hexane. Based on bioactivity tests, we isolated eight fractions of the aqueous MeOH fraction from Astragalus sinicus L. To identify biologically active compounds that inhibit inflammatory responses, we fractioned the polar and non-polar layers of nine types of leguminous plants. From the fractionated samples, we identified that the aqueous methanol layer of Astragalus sinicus L.
exhibited the strongest biological activity measured by the inflammatory response in human keratinocytes induced by cytokines or UVB (data not shown). Therefore, we further purified eight fractions of this plant's aqueous methanol layer (Figure 1a). Test the biological activity of the eight purified fractions of the aqueous methanol layer of Astragalus sinicus L. Although ROS production was not detected in quiescent keratinocytes treated with vehicles, production increased markedly by stimulation with UVB or cytokines.
Among the eight fractions, rf3 and rf4 showed the highest ROS removal activity in keratinocytes induced by UVB or cytokines (Figure 1b and Supplementary Figure S. To determine if Astragalus sinicus L. extracts can regulate certain immune responses, particularly with respect to the differentiation of CD4+ T cells), we examined the expression and production of specific master regulatory factors of T cells and their distinctive cytokines in CD4+ cells. More than 4 T cells in the appropriate polarization conditions.
The mRNA levels of the master regulatory factors of Th2 and Th17 cells, GATA-3 and RORî³t, respectively, were effectively suppressed by treatment with rf3 and rf4, while the mRNA levels of the master regulatory factor of Th1 cells, T-bet, were not affected. In contrast, treatment with rf3 and rf4 significantly increased the mRNA levels of the Treg cell master regulatory factor, Foxp3 (Figure 3a). Next, we examined the effects of Astragalus sinicus L. fractions on the expression of distinctive T-cell-specific cytokines of CD4+ T-cell subsets, 15, 16, 17. The mRNA levels of Th2 and Th17 cytokines, such as IL-4, IL-17A and IL-22, were effectively decreased by treatment with rf3 and rf4, while the mRNA levels of the Th1 cytokine TNF-α, but not IFN-γ, decreased marginally with treatment with rf3 alone.
In contrast, mRNA levels of the anti-inflammatory cytokine IL-10, the characteristic cytokine of Th2 and Treg cells, increased significantly with treatment with rf3, but not with rf4 (Figure 3b). Based on the results of mRNA levels in T-cell-specific master regulatory factors and their distinctive cytokines, the production of the cytokine IL-17A, characteristic of Th17 cells, was effectively inhibited by treatment with rf3 and rf4 under conditions of Th17 polarization. In addition, IL-10 production in Th2 and Treg cells increased significantly with treatment with rf3 and rf4 (Figure 3g). These results suggest that Astragalus sinicus L.The extracts can regulate inflammatory diseases, such as allergy, asthma and systemic autoimmune diseases, induced by an excess of activated Th2 and Th17 cells.
To identify the location of NF-κB p65, one of the proteins of the NF-κB subunit, we performed an immunofluorescence and immunofluorescence analysis. NF-κB p65 was located in the cytosolic region of inactive keratinocytes, but almost all of the protein translocates to the nuclear region after cytokine stimulation. Rf4 effectively inhibited the nuclear translocation of NF-îºB p65, and this inhibitory effect was similar to or much stronger than that of CE or rf3 (figures 5b). and c).
In addition, we examined the NF-κB-dependent promoter activity and discovered that rf4 showed a slightly stronger inhibitory effect than that observed with rf3 or CE on keratinocytes stimulated by cytokines containing a construct of the NF-κB-luciferase luciferase system (Figure 5d). Therefore, rf3 and rf4, present in the aqueous methanol extracts of Astragalus sinicus L. To examine the mRNA levels of proinflammatory mediators in the ears of mice, we performed a quantitative PCR analysis in real time. In accordance with histological changes, the mRNA levels of the main inflammatory cytokines and chemokines known in psoriasis, such as IL-17A, IL-22, CXCL1, CXCL10, CCL17, CCL20, CCL27 and CCR6, increased markedly with the injection of IL-23, as well as in human keratinocytes stimulated by cytokines, while the levels of these mRNAs decreased effectively with rf3 and rf4 (Figure 6b and Supplementary Figure S3c).Along with these cytokines, the mRNA levels of several pro-inflammatory cytokines, including IL-1α, IL-1β, and IL-6, were increased in mouse ears injected with IL-23, while these levels were also effectively decreased with rf3 and rf4 (Supplementary Figure S3b).
In contrast, mRNA levels of the anti-inflammatory cytokine IL-10 increased significantly with treatment with rf3 or rf4 (Figure 6b). The extracts may be able to suppress inflammatory skin diseases, such as psoriasis. To identify biologically active ingredients against inflammatory diseases, we fractioned layers of water, aqueous methanol and n-hexane from nine types of leguminous plants, and examined their antioxidant activities and anti-inflammatory in human keratinocytes. Among the fractions, rf3 and rf4, isolated from the aqueous methanol layer of Astragalus sinicus L.
Benefits were observed to counteract memory disorders in the astragalus due to its activity as a factor promoting nerve growth (1) and its ability to increase the density of the M-cholinergic receptor in senile rats (1). Astragalus polysaccharide (APS) is a compound extracted from the traditional Chinese herb Astragalus membrananum oily, which has several pharmacological properties, improves brain function and immune regulation, and has anti-aging effects. The health benefits of astragalus root may include boosting the immune system and improving kidney or heart function.
