Effects of volatile oil from Acori Graminei Rhizoma on glial fibrillary acidic protein and immediate early genes expressions in the basal lateral amygdala of the inflammatory pain rats / 解剖学报

Objective To construct a rat model of inflammatory pain by injecting complete Freund' adjuvant (CFA) to study effects of volatile oil of Acori Graminei Rhizoma on the expression of glial fibrillary acidic protein (GFAP) and immediate early gene c-fos in the basal lateral amygdale (BLA) of the inflammatory pain rats. Methods Thirty-six adult male SD rats were randomly divided into 6 groups; control group, sham group, CFA group, CFA+ 5 g/( kg · day) volatile oil of Acori Graminei Rhizoma group, CFA+10 g/(kg · day) volatile oil of Acori Graminei Rhizoma group, CFA+20 g/(kg · day) volatile oil of Acori Graminei Rhizoma group, six rats in each group were taken gavage for 21 days. Immunofluorescence and Western blotting methods were used to detect the expressions of GFAP and c-fos in the BLA of all rats. Results Immunofluorescence and Western blotting results showed that compared with the control group, the positive expression of GFAP and c-fos in the BLA of the CFA rats were significantly increased (P<0.01). After treatment of the volatile oil from Acori Graminei Rhizoma, the positive expressions of GFAP and c-fos were reduced compared to the CFA group, as well as the expression levels were decreased in the dose-dependent manner (P<0.01). Compared with the low dose group, the positive expression of GFAP and c-fos of high dose group were decreased significantly (P<0.01). Conclusion The volatile oil fraction from Acori Graminei Rhizoma could reduce the expressions of GFAP and c-fos the BLA of CFA-induced chronic inflammatory pain model rats.

Illuminating the Activated Brain: Emerging Activity-Dependent Tools to Capture and Control Functional Neural Circuits / 神经科学通报·英文版

Immediate-early genes (IEGs) have long been used to visualize neural activations induced by sensory and behavioral stimuli. Recent advances in imaging techniques have made it possible to use endogenous IEG signals to visualize and discriminate neural ensembles activated by multiple stimuli, and to map whole-brain-scale neural activation at single-neuron resolution. In addition, a collection of IEG-dependent molecular tools has been developed that can be used to complement the labeling of endogenous IEG genes and, especially, to manipulate activated neural ensembles in order to reveal the circuits and mechanisms underlying different behaviors. Here, we review these techniques and tools in terms of their utility in studying functional neural circuits. In addition, we provide an experimental strategy to measure the signal-to-noise ratio of IEG-dependent molecular tools, for evaluating their suitability for investigating relevant circuits and behaviors.

Expression of Activating Transcription Factor 3 in Ischemic Penumbra Region Following Focal Cerebral Ischemia / 대한해부학회지

It has been demonstrated that some of immediate early genes (IEGs) such as c-Jun or fos are induced immediately following neuronal injury and they play an important role in determining the fate of the injured neurons. Of IEGs, the activating transcription factor 3 (ATF3) is focused by many investigators, because they are expressed in various types of neural insults and have been known to serve a diverse function in both neuronal survival and death. However, little is known about the functional role of ATF3 in ischemic brain injury. So in this study, the authors examined the expression pattern of the activating transcription factor 3 (ATF3) following middle cerebral artery (MCA) occlusion-reperfusion injury. According to the findings obtained by triphenyltetrazolium chloride (TTC) stains, the authors have classified the infarcted area into two regions, the ischemic core region and the ischemic penumbra region. In both regions, many neurons underwent neuronal degeneration, characterized by the shrunken nuclei with eosinophilic perikaryon. The H & E stain also demonstrated the increased number of probable activated astrocytes and microglia in the ischemic brain regions and this was confirmed by GFAP- and OX42-immunohistochemistry. Immunohistochemical study for ATF3 also demonstrated the specific upregulation of ATF3 in the nuclei of neurons under ischemic injury, but not in those of the contralateral regions. Interestingly, the number of the ATF3 positive neurons in the ischemic penumbra regions outnumbered that of the ischemic core regions. Based on many reports that the neuronal death in ischemic penumbra region is caused by programed cell death rather than by necrosis which is main cause of neuronal death in ischemic core region, our results could suggest that the ATF3 is an important IEGs which determine the fate of the ischemic neurons.

Propofol-induced Immediate Early Gene Expression in Human Neuroblastoma Cell Lines / 대한마취과학회지

BACKGROUND: General anesthetics were known to induce expression of immediate early genes (IEGs), including c-fos and c-jun. However, mechanisms of IEG induction by general anesthetics were not fully understood. METHODS: IEG induction by propofol, a kind of intravenous anesthetics, and signal transduction pathways for propofol-induced IEG expression were investigated in human neuroblastoma cell line IMR32 and CHP134 with Northern and Western blot analysis. RESULTS: Cell viability was significantly decreased in IMR32 and CHP134 treated with increasing concentrations of propofol. IMR32 was more sensitive to propofol-induced cytotoxicity than CHP134. Propofol did not affect the cell cycle profile of IMR32. Expression of cyclin A, cyclin B1, CDK4 and CDK6 was increased in IMR32 by propofol treatment in a time-dependent manner. However, expression of cyclin A and CDK4 was decreased in CHP134. Proliferating cell nuclear antigen (PCNA) was increased in both IMR32 and CHP134 treated with propofol from 6 h to 24 h. c-fos and c-jun were induced by propofol treatment in both cells. Propofol also induced extracellular signal-regulated kinase (ERK) phosphorylation in both cells. Pretreatment of PD98059, an MEK inhibitor, blocked propofol-induced c-fos and c-jun expression.Propofol treatment was decreased nuclear transcription factor-kappa B (NF-kappa B) expression in IMR32, but not in CHP134. CONCLUSIONS: Propofol-induced c-fos expression might be mediated through ERK phosphorylation in both IMR32 and CHP134. Propofol-induced cytotoxicity, changes in expressions of cell cycle regulatory proteins, expression of IEGs, ERK phosphorylation, and NF-kappa B expression were different between IMR32 and CHP134.

Expression of immediate early gene following N-methyl-N-nitrosourea-induced retinal degeneration in SD rats / 中国药理学通报

Aim To study the molecular mechanism of retinal toxicity induced by N-methyl-N-nitrosourea (MNU) in SD rats. Methods A single intraperitoneal injection of 60 mg?kg~(-1) MNU was given to 50-day-old female SD rats. After MNU treatment for different times, the rats were sacrificed and both eyes were enucleated immediately and processed for histological examination. The apoptotic index of photoreceptor cells was calculated by TUNEL labeling and the expression of c-jun and c-fos genes was detected by RT-PCR technique. Results After the application of MNU for 24 h,the disorientation of photoreceptor outer segments was seen. The outer nuclear layer and photoreceptor layer were almost completely lost at 7 d. Apoptosis had already started at 12 h post-MNU and peaked at 24 h. MNU time-dependently up-regulated the expression of c-jun and c-fos genes. Conclusion MNU has a toxic effect on retina by up-modulating expression of c-jun and c-fos genes to promote photorecptor cells apoptosis.

Changes of AP-1 Binding Activities in Rat Brain by Electroconvulsive Shock / 신경정신의학

OBJECTIVES: ECS could have therapeutic effects on psychiatric illnesses by inducing IEGs, which in turn regulates expression of their target genes. We observed AP-1 binding activity and identified AP-1 binding proteins in NMDAR1, late response gene of IEGs, which considered as the candidate gene for schizophrenia. METHODS: By gel shift assay and supershift assay, we observed binding activities and AP-1 binding proteins in NMDAR1. Because IEGs are induced rapidly but transiently by external stimuli, there is a possibility that the expression of IEGs is negatively feedbacked by their own products via their AP-1 binding sites. For that purpose, we also observed AP-1 binding activity of c-fos and c-jun via gel shift and supershift assay. RESULTS: ECS increased AP-1 binding activities of NMDAR1 gene, contributed by c-Fos and its related proteins. Peak of the increased binding was 60 minutes in both hippocampus and cerebellum. Though expression of c-Fos and c-Jun were increased by ECS, there were no changes in AP-1 binding activities after ECS. AP-1 sites of IEGs were binded by CREB, regardless of ECS. CONCLUSION: There is a possibility that ECS induced IEG expression, and then incresed expression of NMDR1 by binding of expressed IEGs to the AP-1 site of NMDAR1. ECS did not increase AP-1 binding activities of IEGs. This suggests that the regulation of IEGs' expression can not be influenced mainly by AP-1 site.

Rapid induction of mRNAs for liver regeneration genes by hepatopoitin and partial hepatectomy / 中华消化杂志

Objective After partial hepatectomy, hepatopoietin(HPO) increase rapidly. The present study was designed to investigate how rhHPO and partial hepatectomy induce the expression of immediate early gene. Methods The Wister rats were divided into hepatectomy group and control group. The different gene expression within 1 h after 2/3 partial hepatectomy and in primary cultured hepatocytes system were examined by using representational difference analysis. Results In the expressed sequence tag(EST) library, most of these genes were identified as immediate early genes, and one new gene Tec that might be associated with liver regeneration in the EST library was found. Moreover, the Tec gene was rapidly induced by 2/3 partial hepatectomy. The expression peak achieved within 1 2 h after operation. HPO can rapidly induce the expression of these genes(c fos, LRF 1 and Tec, etc.) in primarily cultured rat hepatocytes. Conclusion rhHPO and partial hepatectomy can rapidly induce the expression of immediate early gene. This is the first report that Tec gene is an immediate early gene related to liver regeneration.

The Induction of Immediate Early Genes and Phosphorylation of p42, p44 MAPK and Elk-1 by Kainic Acid in Developing Rat Hippocampus / 신경정신의학

OBJECTIVES: In order to investigate the maturational process of intracellular signal transduction system in rat brain, we studied the induction of the immediate early genes(IEGs)c-fos, junB, and TIS1 in each developmental stage after kainic acid(KA)induced seizure in young rat hippocampus and then compared these with the results after electroconvulsive shock(ECS) And to elucidate the induction mechanism of c-fos via mitogen-activated protein kinase(MAPK)by KA in each developmental stage, we investigated the phosphorylation of p42, p44 MAPK and Elk-1 after KA treatment in young rat hippocampus. METHODS: We examined the induction patterns of IEGs by northern blot analysis, and the phosphorylation of p42, p44 MAPK and Elk-1 by immunoblotting in rat hippocampus at post-natal day 7, 14, and 21(P7, P14 & P21) respectively after intraperitoneal injection of KA. RESULTS: Unlike ECS, KA did not induce c-fos, junB, and TIS1 in P7 hippocampus. But these genes were apparently induced at P14 and to an adult level at P21. These three IEGs showed similar temporal patterns of induction in each developmental stage. Although the basal level of phosphorylated 42p, 44p MAPK was considerable in P7 rat hippocampus, the increase of phosphorylation after KA treatment was observed at P14 . While the phosphorylation of Elk-1 was detected with high basal level in P7 rat, the amount of phosphorylated Elk-1 was not changed after KA treatment. CONCLUSION: Our results suggest that the differences in IEGs induction patterns between KA and ECS may be due to the differences in the activated signal transduction pathways. And our results also implicate that the signal transduction system involved in MAPK phosphorylation after KA treatment mature with aging and c-fos induction via MAPK activation may be regulated through some pathways other than Elk-1 in rat hippocampus.

Effect of quercetin combined with Praziquantel on expression of myocardial immediate early gene and transforming growth factor-?1 in mice infected with Schistosomiasis japanica / 中草药

Objective To study the effects and mechanisms of myocardial injury in schistosome-infected mice.To investigate the effects and mechanisms of quercetin on myocardial injury due to the development of hepatic fibrosis after being treated by Praziquantel.Methods Eighty mice were divided into four groups:group A,group B,group C,and group D.Group A,group B,and group C were infected with Schistosoma japonicum cercariae.After 8 weeks,group A was treated with Praziquantel 500 mg/kg for 2 d,group B was treated with quercetin 30 mg/(kg?d) for 8 weeks after being treated with Praziquantel 500 mg/kg for 2 d.Group C was taken as experimental control without any treatment.Group D was taken as normal control.At the week 16,all mice were sacrificed and a part of liver tissue and myocardium tissue were preserved.HE Staining,electric microscope,RT-PCR,and immunohistochemical technique were applied to observing the changes of hepatic and cardiac histopathology,myocardial ultramicrostructure,the expression of myocardial c-fos,c-jun mRNA,and the contents of myocardial transforming growth factor-?1 (TGF-?1),typeⅠand typeⅢcollagen in mice infected with S.japonicum before and after treatments. Results There was different degree of myocardial injury among three groups of experimental control, Praziquantel treatment,Praziquantel combined with quercetin treatment.Praziquantel treatment relieved the degree of hepatic fibrosis and myocardial injury.The content of myocardial c-fos mRNA,c-jun mRNA,TGF-?1,typeⅠand typeⅢcollagen were obviously reduced compared to the experimental control. When Praziquantel treatment combined with quercetin,the degree of hepatic fibrosis and myocardial injury were further relieved.Although the content of myocardial c-fos mRNA,c-jun mRNA,TGF-?1, typeⅠand thypeⅢcollagen were still higher than those in normal control,those were reduced significantly compared to the group treated with Praziquantel.Conclusion Hepatic cirrhosis due to advanced schistosomiasis may lead to cardiac remodeling by stimulating the expression of immediate early gene and promoting the overexpression of TGF-?in myocardium.Anti-fibrosis therapy can reduce the degree of cardiac remodeling.Quercetin may protect myocardium through reducing the degree of hepatic fibrosis and inhibiting the expression of immediate early gene,which could decrease the level of myocardial TGF-?1.

Inhibition of Quercetin on Liver Fibrosis due to Schistosoma japonicum Infection and on the Expression of Immediate Early Gene and Metalloproteinase 1 Inhibitor in Liver Tissue of Mice / 中国寄生虫学与寄生虫病杂志

Quercetin and praziquantel were used to treat mice with hepatic fibrosis due to Schistosoma japonicum infection.Quercetin treatment obviously relieved the degree of hepatic fibrosis,significantly reduced the expression of immediate early gene,tissue inhibitor of metalloproteinase 1(TIMP 1),typesⅠ and Ⅲ collagen compared to the control.The expression of c-jun mRNA,typeⅠ and type Ⅲ collagen were reduced significantly compared to the group treated with praziquantel,whereas no difference in the expression of c-fos mRNA and TIMP1 between the two groups,indicating that quercetin may have better effect on schistosomal liver fibrosis than praziquantel in the long term.