Neuroprotection by Valproic Acid in Mouse Models of Permanent and Transient Focal Cerebral Ischemia

Valproic acid (VPA) is a well-known anti-epileptic and mood stabilizing drug. A growing number of reports demonstrate that VPA is neuroprotective against various insults. Despite intensive efforts to develop new therapeutics for stroke over the past two decades, all treatments have thus far failed to show clinical effect because of treatment-limiting side effects of the drugs. Therefore, a safety-validated drug like VPA would be an attractive candidate if it has neuroprotective effects against ischemic insults. The present study was undertaken to examine whether pre- and post-insult treatments with VPA protect against brain infarct and neurological deficits in mouse transient (tMCAO) and permanent middle cerebral artery occlusion (pMCAO) models. In the tMCAO (2 hr MCAO and 22 hr reperfusion) model, intraperitoneal injection of VPA (300 mg/kg, i.p.) 30 min prior to MCAO significantly reduced the infarct size and the neurological deficit. VPA treatment immediately after reperfusion significantly reduced the infarct size. The administration of VPA at 4 hr after reperfusion failed to reduce the infarct size and the neurological deficit. In the pMCAO model, treatment with VPA (300 mg/kg, i.p.) 30 min prior to MCAO significantly attenuated the infarct size, but did not affect the neurological deficit. Western blot analysis of acetylated H3 and H4 protein levels in extracts from the ischemic cortical area showed that treatment with VPA increased the expression of acetylated H3 and H4 at 2 hrs after MCAO. These results demonstrated that treatment with VPA prior to ischemia attenuated ischemic brain damage in both mice tMCAO and pMCAO models and treatment with VPA immediately after reperfusion reduced the infarct area in the tMCAO model. VPA could therefore be evaluated for clinical use in stroke patients.

Effects of (-)-Epigallocatechin-3-gallate on Brain Infarction and the Activity Change of Matrix Metalloproteinase-9 Induced by Middle Cerebral Artery Occlusion in Mice

Matrix metalloproteinases (MMPs) can degrade a wide range of extracellular matrix components. It has been reported that MMP-9 are activated after focal ischemia in experimental animals. (-)-Epigallocatechin-3-gallate (EGCG), a major constituent of green tea polyphenols, is a potent free radical scavenger and reduces the neuronal damage caused by oxygen free radicals. And it has been known that EGCG could reduce the infarction volume in focal brain ischemia and inhibit MMP-9 activity. To delineate the relationship between the anti-ischemic action and the MMP-9-inhibiting action of EGCG, we investigated the effect of EGCG on brain infarction and the activity of matrix metalloproteinase-9 induced by permanent middle cerebral artery occlusion (pMCAO) in ICR mice. EGCG (40 mg/kg, i.p. 15~30 min prior to MCAO) significantly decreased infarction volume at 24 hr after MCAO. GM 6001 (50 mg/kg, i.p. 15~30 min prior to MCAO), a MMP inhibitor, also significantly reduced infarction volume. In zymogram, MMP-9 activities began to increase at ipsilateral cortex at 2 hr after MCAO, and the increments of MMP-9 activities were attenuated by EGCG treatment. Western blot for MMP-9 also showed patterns similar to that of zymogram. These findings demonstrate that the anti-ischemic action of EGCG in mouse focal cerebral ischemia involves its inhibitory effect on MMP-9.

Effect of immobilization stress on the expression of TH, BDH and CRH gene in rat brain

PURPOSE: Catecholamines are the neuro-transmitters in the sympathetic nervous system (SNS) and are activated by stress stimulus. Tyrosine hydroxylase (TH) and Dopamine-beta-Hydroxylase (DBH) are very important enzymes in the catecholamine synthesis. Corticotropin releasing hormone (CRH) is released in the process of reacting to stresses. The aim of this study is to find out what effects immobilization stresses have on the expression of TH, BDH and CRH mRNA in a rat's brains. METHODS: We compare expression levels in rat's brains of TH, DBH and CRH mRNA induced by immobilization stresses between the test group and controled group. The expression levels of TH, DBH and CRH mRNA are measured by RT-PCR and the Western Blotting Analysis (WBA). RESULTS: In brains and adrenal glands of the immobilization stress group, the expression levels of TH and DBH mRNAs are significantly two to three times higher (P<0.01), and CRH mRNAs are approximately one and a half times higher (P<0.05) than those of controlled group. CONCLUSION: This study suggest that the expression levels of TH, DBH and CRH mRNAs are activated by stress stimulus in a rat's brains and adrenal glands.

Cloning and Expression Pattern of FSTL1 mRNA in Normal Myometrium and Uterine Leiomyoma / 대한산부인과학회잡지

OBJECTIVE: To study the influence and cloning of differentially expressed genes in human female normal myometrium and uterine leiomyoma tissue. METHODS: In this experiment, human uterus tissues (n=25) were taken for total RNA isolation by using Trizol reagent. Differential display was performed by using GeneFishingTM DEG Kit and processed to cDNA sequencing and gene cloning for Follistatin-like 1 (FSTL1). Data were analyzed with the image Master VDS software and statistical significance was defined as p<0.05 by paired t test results. RESULTS: FSTL1 mRNA expression level was significantly higher (p<0.05) in normal and adjacent normal myometrium tissues than uterine leiomyoma tissue of women in the reproductive age. Whereas in the menopausal age, FSTL1 mRNA expression level was significantly higher (p<0.05) in uterine leiomyoma than normal myometrium. There was no significant differences between uterine leiomyoma and adjacent normal myometrium. CONCLUSION: Although the mechanisms of FSTL1 gene were uncertain, FSTL1 seemed to play an important role in the growth of uterine leiomyoma, it also might be related to the regulation of uterine leiomyoma growth inhibiting factors by modulating Follistatin related protein gene (FLRG) system.