摘要
目的 研究中药防风中5-O-甲基维斯阿米醇苷对脂多糖(LPS)诱导的小鼠RAW264.7细胞炎症的保护作用及其作用机制。方法 建立LPS诱导小鼠RAW264.7细胞体外炎症模型,采用MTT比色法测定不同浓度(160、80、40、20、10μmol/L)5-O-甲基维斯阿米醇苷对小鼠RAW264.7细胞活性的影响;实验分空白组,LPS组,阳性对照组(吲哚美辛,10μmol/L),5-O-甲基维斯阿米醇苷低(20μmol/L)、中(40μmol/L)、高(80μmol/L)剂量组,空白组、LPS组加入空白培养基,其余给药组加入相应药物,2h后加入LPS(10μg/mL),采用格里斯试剂法检测小鼠RAW264.7细胞上清液的一氧化氮(NO)的分泌,酶联免疫吸附试验法检测小鼠RAW264.7细胞中的前列腺素E_(2)(PGE_(2))、肿瘤坏死因子α(TNF-α)含量,RT-qPCR检测小鼠RAW264.7细胞核因子-κB(NF-κB)p65、白细胞介素1β(IL-1β)、诱导型一氧化氮合酶(iNOS)、环氧合酶2(COX-2)的mRNA表达,Western blot检测Toll受体4(TLR4)、NF-κB p65蛋白表达。结果5-O-甲基维斯阿米醇苷(10~80μmol/L)对小鼠RAW264.7细胞活性无明显影响。与空白组比较,LPS组小鼠RAW264.7细胞NO、PGE_(2)、TNF-α含量升高(P<0.01),NF-kB p65、IL-1β、INOS、COX-2的mRNA表达及TLR4、NF-κB p65蛋白表达增加(P<0.01);与LPS组比较,5-O甲基维斯阿米醇苷低、中、高剂量组R小鼠AW264.7细胞中NO、PGE_(2)、TNF-α含量均降低(P<0.01),NF-κB p65、IL-1β、INOS、COX-2的mRNA表达和TLR4、NF-κB p65蛋白表达均降低(P<0.01)。结论 防风中5-O甲基维斯阿米醇苷能抑制LPS诱导小鼠RAW264.7细胞炎症反应,具有一定的抗炎作用。
Objective To study the protective effect and mechanism of 4’-O-β-D-glucosyl-5-O-methylvisamminol in Radix Saposhnikoviae on inflammation of mice RAW264.7 cells induced by lipopolysaccharide(LPS).Methods RAW264.7mononuclear macrophages were induced cellular inflammatory model by LPS.The proliferation effect of 4’-O-β-D-glucosyl-5-O-methylvisamminol with different concentrations(160,80,40,20,10 μmol/L) on mice RAW264.7cells was detected by MTT assay.The experiment was divided into blank group,LPS group,positive control group(Indomethacin,10 μmol/L),low(20 μmol/L),medium(40 μmol/L) and high(80 μmol/L) dose groups of 4’-O-β-D-glucosyl-5-O-methylvisamminol.Blank medium was added to blank group and LPS group,and corresponding drugs were added to other dosing groups.LPS(10 μg/ml) was added after 2 h,the secretion of nitric oxide(NO) in supernatant of mice RAW264.7 was detected by Griess reagent method,the contents of prostaglandin E_(2)(PGE_(2)) and tumor necrosis factor α(TNF-α)in the supernatant of mice RAW264.7 cells were determined by enzyme linked immunosorbent assay.The mRNA expressions of nuclear factor κB(NF-κB p65),interleukin-1β(IL-1β),inducible nitric oxide synthase(iNOS) and cyclooxygenase-2(COX-2) were detected by RT-PCR.Toll like receptor 4(TLR4) and NF-κB p65 protein expressions were detected by Western blot Results 4’-O-β-Dglucosyl-5-O-methylvisamminol(10-80 μmol/L) had no effect on RAW264.7 cell viability.Compared with LPS group,the contents of NO,PGE2 and TNF-α in mice RAW264.7 cells were decreased in low,medium and high doses of5-O-methylvisamminol side groups(P<0.01).The mRNA expressions of NF-kB p65,IL-1β,INOS and COX-2 and the protein expressions of TLR4 and NF-kB p65 were decreased(P <0.01).Conclusion 4’-O-β-D-glucosyl-5-Omethylvisammind in Radix Saposhnikoviae inhibit the inflammation of RAW264.7 cells induced by LPS has antiinflammatory effect.
作者
孟莉
郑威
刘蓬蓬
贾天柱
MENG Li;ZHENG Wei;LIU Pengpeng;JIA Tianzhu(Liaoning Traditional Chinese Medicine Institute,Liaoning Province,Shenyang110034,China;College of Pharmacy,Liaoning University of Traditional Chinese medicine,Liaoning Province,Shenyang110032,China)
出处
《中国医药导报》
CAS
2023年第2期16-20,共5页
China Medical Herald
基金
辽宁省教育厅科学研究项目(L201922)。
关键词
防风
5-O-甲基维斯阿米醇苷
RAW264.7
抗炎
Saposhnikovia divaricata
4’-O-β-D-glucosyl-5-O-methylvisamminol
RAW264.7
Anti-inflammatory
