摘要
目的观察丹皮酚是否能够影响巨噬细胞M1极化。方法用Toll样受体7/8激动药雷西莫特(R848)刺激RAW264.7细胞建立M1型巨噬细胞模型。将细胞分为对照组、模型组、高剂量实验组。对照组细胞正常培养;模型组用2μg·mL^(-1)的R848处理24 h,高剂量实验组用2μg·mL^(-1)R848和30μmol·L^(-1)丹皮酚处理24 h。以酶联免疫吸附(ELISA)法检测肿瘤坏死因子-α(TNF-α)分泌水平;以流式细胞术检测CD80阳性表达的巨噬细胞比例;以实时荧光定量多聚核苷酸链反应(Q-PCR)检测TNF-α、白细胞介素-1β(IL-1β)mRNA的表达水平;以蛋白质印迹法检测iNOS蛋白表达情况。结果高剂量丹皮酚干预R848诱导的M1极化24 h后,对照组、模型组和高剂量实验组上清中TNF-α水平分别为(355.55±16.45)、(552.69±18.09)和(447.71±22.71)ng·L^(-1);CD80阳性细胞比例分别为(16.29±0.50)%、(43.57±3.78)%和(36.08±1.40)%;IL-1βmRNA表达水平分别为1.04±0.15、60.02±12.92和34.69±1.75;TNF-αmRNA表达水平分别为1.00±0.04、13.69±0.25和12.20±0.46;iNOS蛋白表达水平分别为1.00±0.00、1.11±0.05和0.91±0.03。模型组上述指标与对照组比较,差异均有统计学意义(均P<0.05);高剂量实验组上述指标与模型组比较,差异均有统计学意义(均P<0.05)。结论丹皮酚能够抑制R848诱导的巨噬细胞M1极化。
Objective To observe whether paeonol could impact the macrophage M1 polarization.Methods The M1 type macrophage model was established by stimulating RAW264.7 cells with Toll-like receptor 7/8 agonist resiquimod(R848).The cells were divided into control group,model group and experimental-H group.The control cells were cultured normally;the model group was treated with 2μg·mL^(-1)R848 for 24 h,the experimental-H group was treated with 2μg·mL^(-1)R848 and 30μmol·L^(-1)paeonol for 24 h.The level of tumor necrosis factor-α(TNF-α)secretion was detected by enzyme linked immunosorbent assay(ELISA);the proportion of macrophages expressing positive CD80 expression was detected by flow cytometry;the expression levels of TNF-αand interleukin-1β(IL-1β)mRNA were detected by real-time fluorescence quantitative polynucleotide chain reaction(Q-PCR);the protein blotting method was used to detect iNOS protein expression was detected by Western blot.Results After 24 h of R848 induced M1 polarization by high dose paeonol intervention,the TNF-αlevels in the supernatants of the control,model and experimental-H groups were(355.55±16.45),(552.69±18.09)and(447.71±22.71)ng·L^(-1),respectively;the proportions of CD80 positive cells were(16.29±0.50)%,(43.57±3.78)%and(36.08±1.40)%,respectively;the expression of IL-1βmRNA were 1.04±0.15,60.02±12.92 and 34.69±1.75,respectively;the expression of TNF-αmRNA were 1.00±0.04,13.69±0.25 and 12.20±0.46,respectively;the expression of iN OS protein were1.00±0.00,1.11±0.05 and 0.91±0.03,respectively.The differences between the model and control groups were statistically significant(all P<0.05);the differences between the high dose experimental group and the model group were statistically significant(all P<0.05).Conclusion Paeonol can inhibit R848 induced M1 polarization of macrophages.
作者
牛玉珍
秦冬儿
金莹莹
唐璠
时乐
梁涛
NIU Yu-zhen;QIN Dong-er;JIN Ying-ying;TANG Fan;SHI Le;LIANG Tao(School of Pharmacy,Nanjing University of Traditional Chinese Medicine,Nanjing 210023,Jiangsu Province,China)
出处
《中国临床药理学杂志》
CAS
CSCD
北大核心
2023年第5期649-653,共5页
The Chinese Journal of Clinical Pharmacology
基金
江苏省研究生实践创新计划基金资助项目(SJCX21-0681)。
