| 1 | Effects of Flower Buds Extract of Tussilago farfara on Focal Cerebral Ischemia in Rats and Inflammatory Response in BV2 Microglia显示文摘Objective: To investigate the effects of the flower buds extract of Tussilago farfara Linné(Farfarae Flos; FF) on focal cerebral ischemia through regulation of inflammatory responses in activated microglia. Methods: Brain ischemia was induced in Sprague-Dawley rats by a transient middle cerebral artery occlusion(t MCAO) for 90 min and reperfusion for 24 h. Twenty rats were randomly divided into 4 groups(n=5 per group): normal, t MCAO-induced ischemic control, t MCAO plus FF extract 300 mg/kg-treated, and t MCAO plus MK-801 1 mg/kg-treated as reference drug. FF extract(300 mg/kg, p.o.) or MK-801(1 mg/kg, i.p.) was administered after reperfusion. Brain infarction was measured by 2,3,5,-triphenyltetrazolium chloride staining. Neuronal damage was observed by haematoxylin eosin, Nissl staining and immunohistochemistry using anti-neuronal nuclei(Neu N), anti-glial fibril ary acidic protein(GFAP), and antiCD11 b/c(OX42) antibodies in ischemic brain. The expressions of inducible nitric oxide synthase(i NOS), tumor necrosis factor(TNF-α), and hypoxia-inducible factor-1 a(HIF-1α) were determined by Western blot. BV2 microglial cel s were treated with FF extract or its main bioactive compound, tussilagone with or without lipopolysaccharide(LPS). Nitric oxide(NO) production was measured in culture medium by Griess assay. The expressions of i NOS, COX-2 and pro-inflammatory cytokines mRNA were analyzed by reverse transcription-polymerase chain reaction. The expression of i NOS, and COX-2 proteins, the phosphorylation of ERK1/2, JNK, and p38 MAPK and the nuclear expression of NF-κB p65 in BV2 cells were determined by Western blot. Results: FF extract significantly decreased brain infarctions in ischemic rats(P<0.01). The neuronal death and the microglia/astrocytes activation in ischemic brains were inhibited by FF extract. FF extract also suppressed i NOS, TNF-α, and HIF-1α expression in ischemic brains. FF extract(0.2 and 0.5 mg/m L, P<0.01) and tussilagone 20 and 50 μmol/L, P<0.01) significantly decreased LPS-induced NO production in BV2 microglia through downregulation of i NOS m RNA and protein expression. FF extract and tussilagone significantly inhibited LPS-induced expression of TNF-α, IL-1β, and IL-6 m RNA, and also suppressed the phosphorylation of ERK1/2, JNK and p38 MAPK and the nuclear expression of NF-κB in a dosedependent manner. Conclusion: FF extract has a neuroprotective effect in ischemic stroke by the decrease of brain infarction, and the inhibition of neuronal death and microglial activation-mediated inflammatory responses. | Ji Hye Hwang Vinoth Kumar R Seok Yong Kang Hyo Won Jung Yong-Ki Park | 2018 | Chinese Journal of Integrative Medicine2018，24，11： | 13 |
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