摘要
目的评价冬凌草甲素对脂多糖(LPS)诱导的小鼠肺泡巨噬细胞(MH-S)的抗炎作用,并探讨其机制。方法台盼蓝法检测不同浓度(0、0.25、0.5、1.0、2.0μg·mL^(-1))冬凌草甲素对MH-S细胞的毒性影响;用荧光定量PCR和酶联免疫吸附试验分别检测不同浓度冬凌草甲素对脂多糖诱导MH-S细胞一氧化氮(NO)、诱导型一氧化氮合酶(iNOS)、环氧合酶-2(COX-2)和前列腺素(PGE2)的mRNA和蛋白质水平的影响;Western blot检测不同浓度冬凌草甲素对脂多糖刺激的MH-S细胞中mitogen-activated protein kinases(MAPK)磷酸化水平的影响。结果台盼蓝实验结果显示不同浓度的冬凌草甲素对MH-S细胞无明显毒性;荧光定量PCR和酶联免疫吸附试验结果显示冬凌草甲素可以剂量依赖性地抑制脂多糖诱导的MH-S细胞中NO、iNOS、COX-2和PGE2 mRNA和蛋白质的表达;Western blot结果显示冬凌草甲素剂量依赖性地抑制p38、ERK1/2和JNK的磷酸化。结论冬凌草甲素能有效抑制脂多糖诱导的MH-S细胞炎症因子的产生,其机制可能与MAPK信号通路的抑制有关。
Objective To investigate the anti-inflammatory roles of oridonin on lipopolysaccharide(LPS)-induced MH-S cells and uncover the underlying mechanism.Methods Trypan blue assay was used to assess the cytotoxicity of different dosage of oridonin(0、0.25、0.5、1.0 and 2.0μg·mL^(-1))on MH-S cells.Cellular inflammation was induced using LPS treatment.The mRNA and protein levels of nitric oxide(NO),inducible nitric oxide synthase(iNOS),cyclooxygenase-2(COX-2)and prostaglandin(PGE2)induced by LPS or LPS and oridonin treatment were measured using enzyme-linked immunosorbent assay(ELISA)and quantitative reverse transcription polymerase chain reaction(qRT-PCR),respectively.Western blot analysis was performed to test the phosphorylation of mitogen activated protein kinases(MAPKs).Results Trypan blue assay showed that oridonin had no cytotoxic effects on MH-S cells.ELISA and qRT-PCR analysis showed that oridonin inhibited the levels of NO,iNOS,COX-2 and PGE2 in LPS-induced MH-S cells with a concentration-dependent manner.Furthermore,oridonin also restrained the LPS-induced the phosphorylation of p38,ERK and JNK in a dose-dependent manner.Conclusion Our data demonstrated that oridonin played protective roles on LPS-induced MH-S cells by reducing proinflammatory mediators via the MAPK pathways.
作者
郭民
张瑞虎
景志杰
庞文彪
陈朝阳
GUO Min;ZHANG Ruihu;JING Zhijie;PANG Wenbiao;CHEN Zhaoyang(Laboratory of Animal Center,Shanxi Medical University,Taiyuan,Shanxi 030001,China)
出处
《石河子大学学报（自然科学版）》
CAS
北大核心
2023年第2期252-257,共6页
Journal of Shihezi University(Natural Science)
基金
山西省科技基础条件平台项目(201805D141008-3)
山西省实验动物专项资金项目(2013-410)。
