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Objective To determine whether four-and-a-half LIM-only protein 2(FHL2)can affect macrophage foa-ming by regulating nuclear factor kappa-B(NF-κB)signaling pathway.Methods FHL2 over-expression plasmids and siRNA of FHL2 were constructed and transfected into human monocyte/macrophages cell line THP-1.Western blot was used to detect the expression of FHL2.The cells were stimulated with oxidized low density lipoprotein(ox-LDL)and the expression of IL-6,IL-1β,TNF-α and other cytokines were detected by ELISA.Oil red O staining was used to detect the degree of cell foaming.The protein expression of NF-κB signaling pathway was detected by Western blot.Results The expression of FHL2 increased after transfected with FHL2 over-expression plasmids while decreased in si-FLH2 transfected cells.FHL2 down-regulated secretion of inflammatory cytokines.Down-regulation of FHL2 allevi-ated THP-1 macrophage foaming.The down-regulation of FHL2 inhibited activation of NF-κB signaling pathway,while the over-expression FHL2 showed an opposite trend.Conclusions FHL2 down-regulation inhibits the activation of NF-κB signaling pathway,reduces the secretion of inflammatory cytokines and alleviates foaming of macrophages.
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Objective To explore the localization of neural specific transcription factor LMO3 mRNA in the central nervous system of the mice.Methods Five male adult mice,weighing 60-80 g,were sacrificed,and the brain and spinal cord were fixed and sliced.In situ hybridization histochemistry staining method with digoxigen-lablled cRNA probe was used in detection of LMO3 mRNA.Results The expression of LMO3 mRNA could be observed in cerebrum,cerebellum,thalamus,brain stem and spine.They were localized in somatic and dendritic profiles at most regions of adult mouse central nervous system.The intense hybridization signal could be seen in Ⅱ-Ⅵ layers of cerebrum,piriform cortex,entorhinal cortex,hippocampus CA1 and dentate gyrus,substantia nigra,motor nucleus of trigeminus,nucleus of hypoglossal,red nucleus,paraventricular nucleus,gigantocellular reticular nucleus,lateral reticular nucleus.The moderate staining was detected in olfactory bulb,hippocampal CA2,amygdala nucleus,cingulated anterior cortex,piriform nucleus etc..The faint signal showed in globus pallidus,spinal nucleus of trigeminal,interposed cerebellar nucleus,ambiguous nucleus,lateral cerebellar nucleus,Purkinje cell of cerebellum,thalamus,caudate putamen,amygdaloid body,nuclei raphe.Conclusion The wide presence of LMO3 mRNA in adult mice central nervous system suggests a role of regulation for this gene in learning,memory and olfaction besides in dopamine neurotransmitter regulation.
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Objective To examine the localization of neuronal specific transcription factor DAT1 mRNA in the central nervous system of the rat. Methods In situ hybridization histochemistry staining method with digoxigen-labeled cRNA probe was used. Results The Transcripts of DAT1 mRNA were localized in somatic and dendritic profiles at most regions of adult rat central nervous system. It can be observed in cerebrum, cerebellum, thalamus, brain stem and spine. The intense hybridization signal can be seen in olfactory bulb, entorhinal cortex, prepirform cortex, striate cortex, hippocampus CA1 and dentate gyrus, lateral dorsal nucleus of thalamus, red nucleus, dorsal tegmental nucleus, central reticular nucleus of medulla oblongata,motor nucleus of trigemina, nucleus of hypoglossal, vagus nerve nucleus, external cuneate nucleus and ambiguous nucleus. The moderate staining was detected in Ⅱ-Ⅵ layer of cerebrum, hippocampus CA2 and CA3, amygdala nucleus, globus pallidus, medial geniculate body, lateral geniculate body, zona incerta, supraoptic nucleus,paraventricular nucleus, arcuate nucleus, substantia nigra, mesencephalon reticular nucleus, reticular nuclei of pons, gigantocellular reticular nucleus, lateral reticular nucleus, medial nucleus of the trapezoid body, vestibular nucleus, dorsal cochlear nucleus, locus ceruleus, cuneate nucleus, nucleus raphes dorsalis. The faint signal showed in septal nuclei, ventral lateral nucleus of thalamus, ventral posterior nucleus of thalamus, posterolateral nucleus of thalamus, dorsomedial nucleus of thalamus, anterodorsal nucleus of thalamus, premammillary nucleus, central gray, superficial gray layer of the superior colliculeus, central nucleus of the inferior colliculeus, lateral nucleus of the inferior colliculeus, trigeminal mesencephalic nucleus, spinal nucleus of trigeminal, nucleus of solitary tract, inferior olivary nucleus, superior central nucleus, cerebellar fastigial nucleus, interposed cerebellared nucleus, lateral cerebellellar nucleus and spinal cord gray matter. Conclusion The results demonstrated the widely presence of DAT1 mRNA in adult rat central nervous system ,close related to the dopaminergic nervous system, suggesting a role of regulation for this gene in various functions of rat central nervous system, especially in dopamine neurotransmitter regulation.