冬凌草甲素通过NF-kapppaB信号通路对LPS刺激树突状细胞的影响  

Effect of Oridonin on LPS-Stimulated Dendritic Cells via NF-κB Pathway

作　　者：贺守第 王健雄 马港圆 王妍洁 李魏明[2] 李一凡 谭宁 黎德育[1] HE Shou-di;WANG Jian-xiong;MA Gang-yuan;WANG Yan-jie;LI Wei-ming;LI Yi-fan;TAN Ning;LI De-yu(Traditional Chinese Medicine Department of Rheumatism,Shenzhen 518052,China;Department of Spine Surgery,Shenzhen 518052,China;Central Laboratory,Huazhong University of Science and Techology Union Shenzhen Hospital,Shenzhen 518052,China;College of Integrated Chinese and Western Medicine,Hunan University of Chinese Medicine,Changsha 410208,China)

机构地区：[1]华中科技大学协和深圳医院中医风湿科,广东深圳518052 [2]华中科技大学协和深圳医院脊柱外科,广东深圳518052 [3]华中科技大学协和深圳医院中心实验室,广东深圳518052 [4]湖南中医药大学中西医结合学院,湖南长沙410208

出　　处：《南昌大学学报（医学版）》2021年第2期13-17,共5页Journal of Nanchang University：Medical Sciences

基　　金：深圳市南山区科技项目(2018033、2019016、2020161)。

摘　　要：目的探讨冬凌草甲素(ORI)通过核因子NF-kappaB(NF-κB)炎症信号通路对脂多糖(LPS)刺激树突状细胞(DC)的影响。方法CCK8法检测ORI对DC活性的影响,并根据CCK8检测法结果,筛选出12.50、25.00μM ORI为干预浓度,实验分为空白组、LPS组、12.50μM ORI+LPS组、25.00μM ORI+LPS组。采用ELISA法检测通路相关炎症因子肿瘤坏死因子-α(TNF-α)及白介素-12(IL-12)浓度,RT-PCR法检测细胞因子白介素-6(IL-6)mRNA的表达,免疫荧光法检测磷酸化(p)-NF-κB p65蛋白(p-NF-κB p65)的表达水平。结果40.00、50.00μM ORI干预DC 24 h后,DC活性降低(P<0.05或P<0.01),50.00μM ORI干预DC 48 h后,DC活性降低(P<0.01);ORI及LPS干预DC 24 h后,与LPS组对比,12.50μM ORI+LPS组、25.00μM ORI+LPS组的TNF-α质量浓度减少(均P<0.01),IL-12的质量浓度差异无统计学意义(P>0.05),IL-6 mRNA的表达下降(P<0.01),显微镜下观察发现12.50μM ORI+LPS组、25.00μM ORI+LPS组细胞内荧光标记p-NF-κB p65的表达减少。结论ORI可能通过抑制NF-κB p65炎症通路的活化从而对LPS刺激DC炎症反应起到抑制作用。Objective To explore the effect of oridonin(ORI)on lipopolysaccharide(LPS)-stimulated dendritic cells(DC)through NF-κB inflammatory pathway.Methods The viability of DC was detected by CCK8 assay.According to the results of CCK8 assay,12.50 and 25.00μM ORI were selected as the intervention concentrations.Cells were divided into four groups:blank group,LPS group,12.50μM ORI+LPS group,and 25.00μM ORI+LPS group.The concentrations of tumor necrosis factor-α(TNF-α)and interleukin-12(IL-12)were measured by ELISA.The expression of interleukin-6(IL-6)mRNA was detected by RT-PCR.The expression of phosphorylated NF-κB p65(p-NF-κB p65)was determined by immunofluorescence assay.Results The viability of DC was significantly decreased by treatment with 40.00 and 50.00μM ORI for 24 h,as well as by 50.00μM ORI intervention for 48 h(P<0.05 or P<0.01).Compared with LPS group,TNF-αconcentration and IL-6 expression de creased in both 12.50μM ORI+LPS group and 25.00μM ORI+LPS group(P<0.01).No obvious change was observed in the level of IL-12(P>0.05).Microscopy showed that the expression of fluorescently labeled p-NF-κB p65 decreased in both 12.50μM ORI+LPS group and 25.00μM ORI+LPS group.Conclusion ORI may suppress the activation of NF-κB p65 pathway,thereby inhibiting the inflammatory response in LPS-stimulated DC.

关 键 词：冬凌草甲素 树突状细胞 NF-kappaB信号通路 

分 类 号：R285.5[医药卫生—中药学]