Study on Effect of Hyacinth Bletilla / Micron Pseudo-ginseng Salve Drug-containing Serum on LPS-induced Inflammatory Injury Model of Macrophages by NF-κB Pathway
XIE Yu,ZHANG Kang,ZHANG Jin-mei
Abstract:
Objective To explore the effect of Hyacinth Bletilla / Micron Pseudo-ginseng salve drug-containing serum on LPS-induced inflammatory injury model of macrophages by NF-κB pathway. Methods The RAW264.7 cells at logarithmic growth stage are processed with lipopolysaccharide (LPS) at different concentrations (5, 10, 15, 20, 25 μg/mL). Its proliferation rate is detected by CCK-8 method. The optimal LPS concentration is determined and the RAW264.7 model of inflammatory injury is established. The cells are divided into normal control group, LPS group and Bletilla / Micron Pseudo-ginseng drug-containing serum + LPS group with different concentration gradient (50, 100, 150, 200, 250, 300 μL/mL). The OD value of each group is detected by CCK-8 method to determine the optimal concentration of drug-containing serum. The cells are divided into normal control group, blank serum group, drug-containing serum group, LPS group, LPS + blank serum group and LPS + drug-containing serum group. The protein expression levels of NF-κB p65 / p-IκBαin cytoplasm and nuclear are determined by Western blotting. Results The optimal concentration of LPS is 10 μg/mL. The optimum concentration of drug-containing serum is 250μL/mL. Compared with the normal control group, the protein expression levels of NF-κB p65 in cytoplasm are significantly decreased in LPS group. The protein expression levels of NFκB p 65 in nuclear and p-IκBα in cytoplasm are significantly increased in LPS group (P<0.01).Compared with the LPS group, the protein levels of NF-κB p65 in nuclear are significantly increased, NF-κB p65 in nuclear and p-IκBα in cytoplasm are significantly decreased in LPS + containing serum group (P<0.01). Conclusion Bletilla / Micron Pseudo-ginseng drug-containing serum inhibits NF-κB p65 entry into the nucleus and IκBα phosphorylation and reduce macrophage secretion of inflammatory cytokines, thereby improving LPS-induced inflammatory reaction by NF-κB signaling pathway.
