Guanxinkang inhibits inflammation and apoptosis and prevents atherosclerosis by regula-ting m6A methylation of FTO
Objective To observe the effects of Guanxinkang on lipopolysaccharide-induced inflammation and apoptosis of hu-man umbilical vein endothelial cells(HUVEC)and the methylation level of demethylase FTO.Methods The cultured HUVEC cells were randomly divided into control group(NC),lipopolysaccharide(LPS)and Guanxinkang(GXK).The effects of LPS and Guanxinkang on the proliferation activity of HUVEC cells were detected by CCK8 method,and the optimal concentration of Guanxinkang and Guanxinkang were selected.After molding and Guanxinkang treatment for24h,m6A RNA methylation level was detected by m6A RNA quantitative kit(colorimetric method),and FTO mRNA and protein expression levels were detected by real-time fluorescence quantitative PCR(RT-qPCR)and Western blotting.The changes of IL-1β,IL-6 and TNF-α inflam-matory factors were detected by ELISA,and apoptosis levels of various cells were detected by flow cytometry.Results(1)The optimal concentrations of LPS and GXK were determined to be 1 μg/mL and 6 μg/mL.(2)Compared with the NC group,the methylation level of m6A RNA increased in the LPS group(P＜0.05),while it decreased in the GXK group compared to the LPS group(P＜0.05).(3)The expression levels of FTO mRNA and protein decreased in the LPS group compared with the NC group(P＜0.05),whereas after GXK intervention,these levels increased compared to the LPS group(P＜0.05).(4)The levels of inflammatory factors IL-1β,IL-6,and TNF-α in the cell supernatant were elevated in the LPS group compared to the NC group(P＜0.05),while they decreased in the GXK group compared to the LPS group(P＜0.05).(5)The apoptotic rate of cells increased in the LPS group compared to the NC group(P＜0.05),whereas it decreased in the GXK group compared to the LPS group(P＜0.05).Conclusion Guanxinkang may inhibit LPS-induced inflammation and apoptosis by regulating demethylase FTO to prevent atherosclerosis.
NETL
NSTL
万方数据
