Inhibitory Effects Of Castrodia Elata Blume On The Inflammatory Process Of Cerebral Ischemic

Objective: Neuroinflammation caused by cerebral ischemia is one of the critical factors leading to aggravation of brain injury. Inhibition of the process of neuroinflammation is expected to be an effective strategy to the relief of the cerebral ischemic injury. Previous studies have shown that a group of anti-inflammation phenolic compounds were extracted from Gastrodia elata Blume and Inhibited the progress of cerebral ischemic injury. However, the relationship between the these compounds-induced anti-cerebral ischemia effects and anti-inflammation effects is still unknown. Therefore, the present study was designed to illustrate the mechanisms of the anti-cerebral ischemic and anti-inflammatory effects induced by the compounds extracted from Gastrodia elata Blume.Methods:1 The effects of anti-inflammatory of phenolic compounds B, C, and D.To investigate the anti-inflammatory effects of compounds B, C and D based on mouse auricle edema model induced by xylene. By using rat planta pedis model induced by injection with carrageenan or AA, the mechanisms of the anti-inflammatory effects of these compounds were identified.2 The effects of anti-cerebral ischemia treated by phenolic compounds B, C and D.The acute cerebral ischemic injury model was built by injecting arachidonic acid via intracarotid artery. Then the anti-inflammatory effects of compounds B, C and D were evaluated based on the neurological deficit scores and the concentration of TNF-α and IL-1β.3 The mechanism involved in the anti-inflammatory of the phenolic compounds B, C and D.3.1 The cell signaling pathways involved.Based on rat acute cerebral ischemic injury model, ELISA kits were used to detect the concentrations of TXA2, 6-keto-PGF1α, LTB4 and Cys LTs in brain which belongs to the metabolites of COX.3.2 Inhibitory effect on the abnormal activation of BV2.MTT methods were usd to compare the difference in BV-2 cells betweeb the control group and the compounds C and D treated group. The concentration of NO in supernatants was measured by Nitrate reductase method. Menwhile, ELISA methods was used to detect the amounts of TNF-α, L-1β, LTB4 and Cys LTs in supernatants. Western blotting analysis was performed to determine the protein expression of 5-LO in BV-2 cells.Results:1 The anti-inflammatory effects of phenolic compounds B, C, and D.All 3 compounds inhibited the mouse auricle edema at the concentration of 20 or30mg/kg. Compound B( 20mg/kg) inhibited rat metatarsal swelling induced by carrageenan which lasted 4 h with average 42.56% inhibitory rate. However,compound B(20mg/kg) only inhibited the rat metatarsal swelling induced by AA within 1 to 2 h. Although compound C and D did not show inhibitory effect on the rat metatarsal swelling induced by carrageenan, these two compounds inhibited rat metatarsal swelling induced by AA with 33.5% and 31.7% average inhibitory rate.2 The anti-cerebral ischemic effects of phenolic compounds B, C and D.Phenolic compound B( 20mg/kg) reduced neurological deficit scores. The tendency of neurological deficit scores was improved by treated with compound C.All the 3 compounds resulted in the reductions of TNF-αand IL-1β concentrations in brain tissue with acute cerebral ischemia injury.3 Mechanism involved in the anti-inflammatory effect of phenolic compounds B, C and D.3.1 The cell signaling pathways involved.Phenolic compounds B significantly reduced the concentration of TXA2 and6-keto-PGF1α but showed no effects on LTB4 and Cys LTs concentrations in the injuredbrain tissue. In contrast, phenolic compounds C and D had no significant effect on the concentrations of TXA2 and 6-keto-PGF1α but markedly reduced the concentration of LTB4 and Cys LTs.3.2 Inhibitory effect on the abnormal activation of BV2.Phenolic compounds C and D(50μg/m L) significantly reduced the NO concentration in BV-2 cells. In addition, compound C depressed the expression of TNF-α and IL-1β. Compound D reduced the release of TNF-α, and reduced IL-1β and50μg/m L. Compounds C and D depressed the expression of LTB4 and Cys LTs of5-LO pathway but had no effect on the protein expression of 5-LO in BV-2cells.Conclusions:1 Phenolic compounds B, C and D inhibit inflammatory reaction in different degree. Compound B acts on the COX pathway, while compounds C and D affect5-LO pathway.2 Phenolic compounds B, C and D attenuate the acute cerebral ischemic injury in brain through reducing the brain inflammatory reaction.3 Phenolic compound B inhibits the brain inflammation by reducing the concentrations of TXA2 and 6-keto-PGF1α of COX pathway. Phenolic compounds C and D inhibit the brain inflammation by depressing the expression of LTB4 and Cys LTs of 5-LO pathway.4 Phenolic compounds C and D control the neuroinflammation by inhibiting BV-2 cell abnormal activation and inhibit the expressions of LTB4 and Cys LTs of5-LO pathway.