Anti-inflammatory Effect of Pueraria lobata (Willd.) Ohwi and P. thomsonii Benth. Polysaccharides on LPS-Induced RAW264.7 Cells
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Abstract:
The anti-inflammatory effects of Pueraria crude polysaccharide (FGC), Pueraria crude polysaccharide (GGC), and Pueraria homogenized polysaccharide (FGJ) were compared in an LPS-induced RAW264.7 cell model. The effects of FGC, GGC, and FGJ on cell viability were detected using a CCK8 kit. Secretion of nitric oxide, TNF-α, and IL-6 in cells was detected using their corresponding kits. The expression levels of NF-κB and Nrf2/HO-1 pathway related proteins were detected using western blot, mRNA expression levels of TNF-α and IL-6 were detected by qPCR, and ROS production was detected using flow cytometry. The results showed that FGC, GGC, and FGJ had no toxicity to cells at concentrations of 10, 20, and 40 μg/mL (p>0.05). FGC and FGJ did not inhibit LPS-induced nitric oxide, TNF-α, and IL-6 secretion (p>0.05) at any concentration. At 40 μg/mL, GGC inhibited nitric oxide, TNF-α, and IL-6 levels by 38.46%, 52.90%, and 55.87%, respectively, which was able to inhibit intracellular ROS release. Inhibition rates reached 37.28%, 41.05%, and 54.09% at concentrations of 10, 20, and 40 μg/mL. The inhibition rates of iNOS, COX-2, p-p65, and p-IκBα were 29.27%, 95.04%, 27.18%, and 28.45% at 40 μg/mL, which was able to promote the expression of Nrf2 and HO-1 proteins. At 40 μg/mL, mRNA expression levels of IL-6 and TNF-α were significantly inhibited, with inhibition rates reaching 57.70% and 81.68%, respectively. The results showed that GGC had a more distinct anti-inflammatory activity than FGC and FGJ, and its anti-inflammatory effect was realized through the NF-κB and Nrf2/HO-1 pathways.
