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Effects of 3,4-oxo-isopropylidene Shikimic Acid (ISA) on the Inflammatory Reaction in Rats Subjected to Middle Cerebral Artery Ischemia and Reperfusion and Its Probable Mechanisms

Author: SunWenYan
Tutor: SunJianNing
School: Beijing University of Traditional Chinese Medicine
Course: Chinese and Western medicine combined with the basis
Keywords: Isopropylidene shikimic acid Cerebral ischemia and reperfusion Inflammation Adhesion molecule Cytokines Cyclooxygenase Transcription factor
CLC: R285.5
Type: PhD thesis
Year: 2005
Downloads: 276
Quote: 0
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Abstract


Ischemic cerebrovascular disease is the first cause of death in China before the six major diseases killer depth to explore the pathogenesis of important significance. In recent years, studies have shown that inflammatory mechanisms in cerebral ischemia / reperfusion injury occurred plays an important role in the development process, the anti-inflammatory treatment of ischemic cerebrovascular disease has become one of the means of intervention. The histological features of cerebral ischemia inflammation leukocyte infiltration, cytokines, adhesion molecules, inflammatory mediators and transcription factors in the pathological process plays a key role. Cytokines, adhesion molecules, transcription factors, and the leukocyte infiltration displayed between the chain reaction, namely, what is called the inflammatory cascade. Synthetic derivative of the active ingredients plant star anise extract shikimic acid (shikimic acid, SA) isopropylidene shikimic acid (3,4-oxo-isopropylidene shikimic acid, ISA) is from Chinese medicine octagonal case, the study showed that ISA can significantly inhibit a variety of inducers induced platelet aggregation and experimental thrombus formation, inhibition of vascular endothelial cell activation, with good protection against focal cerebral ischemia. In this paper, the enzyme-linked immunoassay, radioimmunoassay, immunohistochemistry, reverse transcriptase polymerase chain reaction and other means, from the destruction of the blood-brain barrier, the generation of cerebral edema, leukocyte infiltration, cytokine expression of adhesion molecules, epoxy aggregate enzyme activation and activation of transcription factors and other inflammatory aspects of ISA intervention rat transient middle cerebral artery occlusion (tMCAO) model of inflammatory response and the possible mechanism of action. The protective effects Objective: To study 1 isopropylidene shikimic acid on brain ischemia-reperfusion in rat neurons ISA cerebral ischemia reperfusion neurological symptoms in rats, infarct size, serum NSE levels and brain pathomorphology . Method: Refer to Koizumi and Nagasawa method produced tMCAO rat model, according to Zea Longa 5 level score after 3 h, and neurological symptoms score in 6 h and 24 h, 24 h after serum were measured by radioimmunoassay neural elements specific enolase (NSE) content, infarct size measured with TTC staining, HE staining pathological morphology. Results: tMCAO rats had obvious symptoms of nerve defects and massive cerebral infarction, serum NSE levels significantly increased neuron degeneration and necrosis or demyelination, interstitial edema, leukocyte infiltration. ISA 200 mg / kg, ISA 100 mg / kg can significantly reduce the rat model of neurological symptoms score and serum NSE levels, reduce infarct relieve pathological damage brain tissue. Conclusion: ISA can be reduced to the neurons in ischemic injury. Isopropylidene shikimic acid on brain ischemia-reperfusion intervention role of the inflammatory response in rats Objective: To investigate the ISA tMCAO blood-brain barrier, cerebral edema, leukocyte infiltration. : Evans Blue, IgG permeability experimental determination of cerebral microvascular permeability change reflects the destruction of brain microvascular matrix metalloproteinase -9 (MMP-9) were measured by immunohistochemistry, wet and dry weight was measured brain changes in tissue water content reflects the infiltration of leukocytes to the brain tissue myeloperoxidase (MPO) activity was measured. Results: tMCAO rat brain microvascular Iraq, Evans Blue and IgG showed significant permeability of MMP-9 expression of; significantly increased brain water content, MPO activity was significantly enhanced. ISA 200 mg / kg, ISA 100 mg / kg can significantly reduce the tMCAO rat brain microvascular permeability, content of Evans blue and IgG expression was significantly reduced; ISA 200 mg / kg can significantly reduce the 2 isopropylidene shikimic acid resistant inflammatory mechanism of cerebral ischemia reperfusion injury in cerebral microvascular MMP-9 positive area and integral optical density; the ISA 200 mg / kg, the ISA 100 mg / kg can significantly reduce the tMCAO rat brain water content and MPO activity, improve cerebral edema and reduce leukocyte infiltration. Conclusion I: SA can significantly reduce cerebral ischemia-reperfusion-induced inflammatory response. Objective: To observe the isopropylidene shikimic acid on cerebral ischemia reperfusion rat adhesion molecule expression tMCAO rat brain tissue intercellular adhesion molecule -1 (ICAM-1), lymphocyte function-associated antigen-1 ( LFA-1), macrophage antigen complex -1 (Mac-1) expression of the changes, and the ISA. Method: cerebral ischemia and reperfusion 21 h 3 h tMCAO rat with paraformaldehyde heart perfusion brain, paraffin-embedded sections of ICAM-1 of LFA-1 and Mac-1 immunohistochemical staining, observed under the microscope and image analysis. Results: cerebral ischemia reperfusion ICAM-1 is expressed in a large number of brain microvascular endothelial cells, LFA-1 and Mac-1 expression of leukocyte infiltration in the brain tissue. ISA 200 mg / kg can significantly reduce the number of rat model of ICAM-1 positive expression of vascular positive area and integral optical density and significantly reduce the LFA-1 positive area, integrated optical density and Mac-1 integrated optical density. Conclusion: ISA significant expression of adhesion molecules down after cerebral ischemia and reperfusion. Objective: Research ISA isopropylidene shikimic acid on cerebral ischemia reperfusion rat inflammatory cytokine expression tMCAO rat brain tissue interleukin-1β (IL-1β), interleukin-8 (IL-8), tumor necrosis factor-α (TNF-α), the content and the expression of IL-8 mRNA. Method: cerebral ischemia and reperfusion 21 h 3 h, making brain homogenates, which of IL-1β, IL-8, TNF-α content of the supernatant was determined by ELISA; take the ischemic area of ??brain tissue by RT- PCR was used to study the expression of IL-8 mRNA. Results: tMCAO rat brain IL-1β, IL-8, the content of TNF-α, and IL-8 mRNA expression was significantly increased, the ISA 200 mg / kg and 100 mg / kg can significantly reduce rat brain IL-1β , IL-8, TNF-α level of ISA 200 mg / kg were significantly reduced the expression of IL-8mRNA. Conclusions: ISA can significantly reduce the high expression of inflammatory cytokines after cerebral ischemia and reperfusion. 5 isopropylidene shikimic acid on cerebral ischemia and reperfusion in the rat cyclooxygenase expression Objective: To study the content of the ISA on the tMCAO rat plasma prostaglandin E2 (PGE2), and brain tissue cyclooxygenase -2 (COX -2) protein and mRNA expression. Method: cerebral ischemia and reperfusion 21 h 3 h, rat carotid artery blood, separated plasma PGE2 content was measured by radioimmunoassay; paraffin sections of brain tissue, COX-2 protein expression observed by immunohistochemical methods; take ischemic area of ??brain tissue to study COX-2 mRNA expression by RT-PCR. Results: three in the sham group was expressed at low levels tMCAO in rat plasma PGE2 content was significantly higher brain tissue COX-2 protein

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