When we evaluated changes of glial fibrillary acidic protein (GFAP) and two glutamate transporter (GTs) by immunohistochemistry, expression of GFAP showed a significant increase in complete Freund's adjuvant (CFA)-injected rats; however, this expression was strongly inhibited by electroacupuncture (EA) stimulation. Robust downregulation of glutamate-aspartate transporter (GLAST) and glutamate transporter-1 (GLT-1) was observed in CFA-injected rats; however, EA stimulation resulted in recovery of this expression. Double-labeling staining showed co-localization of a large proportion of GLAST or GLT-1 with GFAP. Using Western blot, we confirmed protein expression of two GTs, but no differences in the mRNA content of these GTs were observed. Because EA treatment resulted in strong inhibition of CFA-induced proteasome activities, we examined the question of whether thermal sensitivities and GTs expression could be regulated by proteasome inhibitor MG132. CFA-injected rats co-treated with EA and MG132 showed a significantly longer thermal sensitivity, compared with CFA-injected rats with or without MG132. Both EA and MG132 blocked CFA-induced GLAST and GLT-1 downregulation within the spinal cord. These results provide evidence for involvement of GLAST and GLT-1 in response to activation of spinal astrocytes in an EA antinociceptive effect. Antinociceptive effect of EA may be induced via proteasome-mediated regulation of spinal GTs.
Two mechanisms of radiation-induced cell death in K562 cells, mitotic catastrophe and apoptosis, are regulated through distinct pathways, mitochondria and caspase-independent and -dependent, respectively. The findings of this study may provide new insights into improving the efficiency of radiotherapy in CML patients.
We explored the neuroprotective effects of a hexane extract from Uncaria sinensis (HEUS) against glutamate-induced toxicity focusing on its anti-apoptotic mechanism in primary cultured cortical neurons. Pretreatment with HEUS resulted in significantly reduced glutamate-induced toxicity in a dose-dependent manner with a decrease in the release of lactate dehydrogenase. Morphological assay and flow cytometry were performed for determination of the type of cell death; according to the results, treatment with HEUS resulted in a significant reduction of glutamate-induced apoptotic cell death. We examined the anti-apoptotic mechanism of HEUS; treatment with HEUS resulted in markedly decreased expression of death receptor (DR)4, which was induced by glutamate stimulation. In contrast, treatment with HEUS resulted in significantly enhanced levels of expression of glutamate-attenuated XIAP and Bcl-2, as well as marked blockade of glutamate-induced Bid cleavage, which inhibits both extrinsic and intrinsic apoptosis pathways. In addition, pretreatment with HEUS resulted in almost complete blockade of glutamate-induced activation of caspases-8, -9 and -3, as well as cleavage of poly (ADP-ribose) polymerase. These findings suggest that the neuroprotective effects of HEUS against glutamate-induced toxicity occur via inhibition of DR4 and expression of anti-apoptotic proteins XIAP and Bcl-2 resulting in effective abrogation of the activation of caspase cascades and promotion of cell survival.
Abstract. The active fraction of the extract of Asiasari radix, the 60% methanol chromatographic fraction from the ethyl acetate layer (PPAR), was used to investigate the melanogenesis signal pathway in B16F10 melanoma cells. PPAR led to significantly decreased melanin synthesis and tyrosinase activity in a dose-dependent manner. PPAR also reduced the expression of melanogenesis-related proteins including microphthalmia-associated transcription factor (MITF), tyrosinase, and tyrosinase-related protein (TRP)s while down-regulating the expression of mRNA of MITF and tyrosinase. Melanogenesis-regulating signals, such as mitogen-activated protein kinase (MEK)/extracellular signalregulated kinase (ERK), phosphatidylinositol 3-kinase (PI3K)/Akt and p38 mitogen-activated protein kinase (MAPK) were evaluated, and ERK was activated by PPAR. The selective inhibitor of MEK/ERK, PD98059, abrogated all suppressive activities of PPAR on melanin biosynthesis, tyrosinase activation and expression of melanogenesis-related proteins. These results suggest that ERK activation by PPAR contributes to reduced melanin synthesis via ERK signal pathway-mediated suppression of MITF and its downstream signal pathway.
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