Z-ligustilide attenuates lipopolysaccharide-induced proinflammatory response via inhibiting NF-KB pathway in primary rat microglia  

作　　者：Jing WANG Jun-rong DU Yu WANG Xi KUANG Cheng-yuan WANG 

机构地区：[1]Department of Pharmacology and Biopharmaceutics, Key Laboratory of Drug Targeting and Drug Delivery Systems Ministry of Education, West China School of Pharmacy, Sichuan University, Chengdu 610041, China [2]Research Center of Medical Chemistry & Chemical Biology, Chongqing Technology and Business University, Chongqing 400067, China

出　　处：《Acta Pharmacologica Sinica》2010年第7期791-797,共7页中国药理学报（英文版）

摘　　要：Aim： To investigate the anti-inflammatory effect of Z-ligustilide （LIG） on lipopolysaccharide （LPS）-activated primary rat microglia. Methods： Microglia were pretreated with LIG I h prior to stimulation with LPS （1 pg/mL）. After 24 h, cell viability was tested with MTT, nitric oxide （NO） production was assayed with Griess reagent, and the content of tumor necrosis factor-α （TNF-α）, interleukin-1β （IL-1β）, and monocyte chemoattractant protein （MCP-1） was measured with ELISA. Protein expression of the nuclear factor-KB （NF-KB） p65 subunit, cyclooxygenase-2 （COX-2）, and inducible nitric oxide synthase （iNOS） was detected with immunocytochemistry I h or 24 h after LPS treatment. Results： LIG showed a concentration-dependent anti-inflammatory effect in LPS-activated microglia, without causing cytotoxicity. Pretreatment with LIG at 2.5, 5, 10, and 20 pmol/L decreased LPS-induced NO production to 75.9%, 54.4%, 43.1%, and 47.6% （P〈0.05 or P〈0.01）, TNF-α content to 86.2%, 68.3%, 40.1%, and 39.9% （P〈0.01, with the exception of 86.2% for 2.5 pmol/L LIG）, LI-1β content to 31.5%, 27.7%, 0.6%, and 0% （P〈0.01）, and MCP-1 content to 84.4%, 50.3%, 45.1%, and 42.2% （P〈0.05 or P〈0.01）, respectively, compared with LPS treatment alone. LIG （10 μmol/L） significantly inhibited LPS-stimulated immunoreactivity of activated NF-KB, COX- 2, and iNOS （P〈0.01 vs LPS group）. Conclusion： LIG exerted a potent anti-inflammatory effect on microglia through inhibition of NF-κB pathway. The data provide direct evidence of the neuroprotective effects of LIG and the potential application of LIG for the treatment of the neuroinflammatory diseases characterized by excessive microglial activation.Aim： To investigate the anti-inflammatory effect of Z-ligustilide （LIG） on lipopolysaccharide （LPS）-activated primary rat microglia. Methods： Microglia were pretreated with LIG I h prior to stimulation with LPS （1 pg/mL）. After 24 h, cell viability was tested with MTT, nitric oxide （NO） production was assayed with Griess reagent, and the content of tumor necrosis factor-α （TNF-α）, interleukin-1β （IL-1β）, and monocyte chemoattractant protein （MCP-1） was measured with ELISA. Protein expression of the nuclear factor-KB （NF-KB） p65 subunit, cyclooxygenase-2 （COX-2）, and inducible nitric oxide synthase （iNOS） was detected with immunocytochemistry I h or 24 h after LPS treatment. Results： LIG showed a concentration-dependent anti-inflammatory effect in LPS-activated microglia, without causing cytotoxicity. Pretreatment with LIG at 2.5, 5, 10, and 20 pmol/L decreased LPS-induced NO production to 75.9%, 54.4%, 43.1%, and 47.6% （P〈0.05 or P〈0.01）, TNF-α content to 86.2%, 68.3%, 40.1%, and 39.9% （P〈0.01, with the exception of 86.2% for 2.5 pmol/L LIG）, LI-1β content to 31.5%, 27.7%, 0.6%, and 0% （P〈0.01）, and MCP-1 content to 84.4%, 50.3%, 45.1%, and 42.2% （P〈0.05 or P〈0.01）, respectively, compared with LPS treatment alone. LIG （10 μmol/L） significantly inhibited LPS-stimulated immunoreactivity of activated NF-KB, COX- 2, and iNOS （P〈0.01 vs LPS group）. Conclusion： LIG exerted a potent anti-inflammatory effect on microglia through inhibition of NF-κB pathway. The data provide direct evidence of the neuroprotective effects of LIG and the potential application of LIG for the treatment of the neuroinflammatory diseases characterized by excessive microglial activation.

关 键 词：Z-ligustilide （LIG） MICROGLIA neuroinflammation LIPOPOLYSACCHARIDE NF-κB tumor necrosis factor-o INTERLEUKIN-1Β iNOS CYCLOOXYGENASE-2 

分 类 号：Q813.11[生物学—生物工程] S942.5[农业科学—水产养殖]