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1.
Acta Physiologica Sinica ; (6): 733-739, 2016.
Article in Chinese | WPRIM | ID: wpr-331609

ABSTRACT

The purpose of the present study was to investigate the effect of advanced glycated albumin (AGE-alb) on the activation of caspase-12, a key molecule in endoplasmic reticulum stress (ERS)-associated apoptotic pathway, and to elucidate the underlying molecular mechanisms of macrophage apoptosis. RAW264.7 macrophages were treated with AGE-alb (2, 4 and 6 g/L), control albumin (C-alb, 4 g/L), tunicamycin (TM, 4 mg/L), or pretreated with 4-phenylbutyric acid (PBA, 5 mmol/L) for 1 h and then treated with AGE-alb (4 g/L). After incubation for 24 h, the cell viability and apoptosis were determined by using MTT assay and TUNEL detection kit, respectively. Lactate dehydrogenase (LDH) activity in media was determined by using an assay kit. The protein levels of caspase-12 were examined by Western blot analysis. The results showed that like TM (an ERS inducer), incubation with AGE-alb led to significant decrease in viability and increase in LDH activity in media and apoptotic rate in a dose-dependent manner. In addition, AGE-alb induced activation of caspase-12 especially at the concentration of 4 and 6 g/L (P < 0.01), which was similar to TM. However, PBA (an ERS inhibitor) protected RAW264.7 macrophages from AGE-alb-induced decrease in viability and increases in LDH activity and apoptosis. Moreover, PBA also inhibited the caspase-12 activation induced by AGE-alb (P < 0.05). These results suggest that AGE-alb may induce apoptosis in RAW 264.7 macrophages, and the mechanism may be related to the activation of ERS-associated apoptotic pathway mediated by caspase-12.


Subject(s)
Animals , Mice , Apoptosis , Caspase 12 , Cell Line, Tumor , Cell Survival , Endoplasmic Reticulum Stress , Macrophages , Phenylbutyrates , Serum Albumin , Tunicamycin
2.
Acta Physiologica Sinica ; (6): 47-54, 2013.
Article in Chinese | WPRIM | ID: wpr-333136

ABSTRACT

The purposes of the present study were to investigate the inhibitory effect of quercetin (QUE) preconditioning on endoplasmic reticulum stress (ERS) inducer tunicamycin (TM)-induced apoptosis in RAW264.7 macrophages and the underlying molecular mechanisms. RAW264.7 cells were pretreated with different concentrations (20, 40, and 80 μmol/L) of QUE for 30 min and then treated with TM (5 mg/L) for 12 h. Cell viability and apoptosis were determined using MTT assay and Annexin V-FITC apoptosis detection kit, respectively. The nuclear translocation of activating transcription factor 6 (ATF6) in cells was detected by immunofluorescence analysis and Western blot. Protein and mRNA expressions of C/EBP homologous protein (CHOP) and Bcl-2 were examined by Western blot and real-time PCR, respectively. The results showed that TM reduced cell viability and induced apoptosis in RAW264.7 macrophages. The cytotoxic effects of TM were significantly inhibited by QUE pretreatment at the concentrations of 40 and 80 μmol/L. Interestingly, we found that QUE also significantly suppressed the TM-induced translocation of ATF6, an ERS sensor, from the cytoplasm to the nucleus. In addition, exposure of RAW264.7 macrophages to TM resulted in a significant increase of the expression of CHOP, a transcription factor regulated by ATF6 under conditions of ERS, as well as a decrease of Bcl-2 at transcript and protein levels. QUE blocked these effects in a dose-dependent manner. These data indicate that QUE can protect RAW264.7 cells from TM-induced apoptosis and that the mechanism at least partially involves its ability to inhibit the ATF6-CHOP signaling pathway.


Subject(s)
Animals , Mice , Activating Transcription Factor 6 , Metabolism , Apoptosis , Cell Survival , Endoplasmic Reticulum Stress , Macrophages , Cell Biology , Quercetin , Pharmacology , Transcription Factor CHOP , Metabolism , Tunicamycin , Pharmacology
3.
National Journal of Andrology ; (12): 953-956, 2011.
Article in Chinese | WPRIM | ID: wpr-239051

ABSTRACT

<p><b>OBJECTIVE</b>To investigate the effects of Compound Xuanju Capsule on the levels of sex hormones and the weight of sexual organs in castrated male rats.</p><p><b>METHODS</b>A randomized model control trail was performed in 60 young male SD rats of SPF grade, of which 12 were included in the normal control group, and the others were castrated and randomly divided into a model control group and a high-dose, a median-dose and a low-dose Xuanju group. The control groups received intragastric administration of normal saline, and the model groups solution of Compound Xuanju Capsule, all for 20 days. Then we determined by radioimmunoassay the levels of testosterone (T), luteinizing hormone (LH) and follicle stimulating hormone (FSH) in the peripheral blood of the rats, and measured the weights of the epididymis, preputial gland, seminal vesicle, prostate and levator ani muscle.</p><p><b>RESULTS</b>The T levels were remarkably lower in the castrated groups than in the normal controls, and significantly higher in the three Xuanju groups than in the model controls (P < 0.01). Both LH and FSH levels were increased in the model control and Xuanju groups as compared with the normal control group, the former with statistically significant difference (P < 0.05) and the latter without. In comparison with the normal controls, the model control rats showed a marked reduction in the indexes of the preputial gland, seminal vesicle, prostate and levator ani muscle, and the high-dose Xuanju group exhibited a significant increase in the seminal vesicle index as compared with the model controls (P < 0.05). There were no statistically significant differences in the indexes of preputial gland, prostate and levator ani muscle among different dose groups (P > 0.05).</p><p><b>CONCLUSION</b>Compound Xuanju Capsule can elevate T and LH levels in the peripheral blood of male SD rats and improve the indexes of their sex organs, which may be an important mechanism behind its effect on ED.</p>


Subject(s)
Animals , Male , Rats , Body Weight , Drugs, Chinese Herbal , Pharmacology , Follicle Stimulating Hormone , Metabolism , Gonadal Steroid Hormones , Metabolism , Luteinizing Hormone , Metabolism , Orchiectomy , Organ Size , Random Allocation , Rats, Sprague-Dawley , Seminal Vesicles , Testosterone , Metabolism
4.
China Journal of Chinese Materia Medica ; (24): 534-536, 2008.
Article in Chinese | WPRIM | ID: wpr-284449

ABSTRACT

<p><b>OBJECTIVE</b>To study the effects of beta-asarone on expression of immediately early gene c-fos in kindling epilepsy rat brain.</p><p><b>METHOD</b>The rats were randomly divided in to beta-asarone groups (200, 100, 50 mg x kg(-1) x d(-1)), difetoin control group (36 mg x kg(-1)) and model group. The remedy was administered orally. The effects were observed in kindling epilepsy model induced by penicillin, then the expression of c-fos were determined by western blot (hippocampus) and immunohistochemical techniques (cortex).</p><p><b>RESULT</b>Beta-asarone could significantly increase the expression of c-fos in kindling epilepsy rat brain, and show its quantity-effect relation. The expression of c-fos in hippocampus was (1139.45 +/- 155.56), (1109.56 +/- 134.03), (1103.73 +/- 235.82) CNT x mm2 in beta-asarone groups, 920.54 +/- 203.20 in model control group, and 1106.26 +/- 186.24 in difetoin group, respectively. The number of c-fos positive cell was 87.1 +/- 2.2, 76.3 +/- 1.3 and 59.9 +/- 1.3 in beta-asarone groups, 39.3 +/- 2.6 in model control group, and 95.2 +/- 1.1 in difetoin group, respectively.</p><p><b>CONCLUSION</b>Beta-asarone can obviously increase the expression of c-fos in epilepsy rat brain. It is one of important response to epilepsy.</p>


Subject(s)
Animals , Female , Male , Rats , Anisoles , Pharmacology , Blotting, Western , Brain , Metabolism , Epilepsy , Drug Therapy , Metabolism , Gene Expression , Immunohistochemistry , Proto-Oncogene Proteins c-fos , Metabolism , Random Allocation , Rats, Sprague-Dawley
5.
Journal of Zhejiang University. Science. B ; (12): 88-97, 2007.
Article in English | WPRIM | ID: wpr-309031

ABSTRACT

Appressorium is an infection structure of the phytopathogenic fungus Magnaporthe grisea. Analysis of gene expression profiles of appressorium development provides insight into the molecular basis of pathogenicity and control of this fungal plant disease. A cDNA array representing 2927 unique genes based on a large EST (expressed sequence tag) database of M. grisea strain Y34 was constructed and used to profile the gene expression patterns at mycelium and appressorium maturation stages. Compared with mycelia, 55 up-regulated and 22 down-regulated genes were identified in mature appressoria. Among 77 genes, 16 genes showed no similarity to the genome sequences of M. grisea. A novel homologue of peptidyl-prolyl cis-trans isomerase was found to be expressed at low-level in mature appressoria of M. grisea. The results indicated that the genes such as pyruvate carboxylase, phospholipid metabolism-related protein and glyceraldehyde 3-phosphate dehydrogenase involved in gluconeogenesis, lipid metabolism and glycolysis, showed differential expression in mature appressoria. Furthermore, genes such as PTH11, beta subunit of G protein and SGT1 involved in cell signalling, were expressed differentially in mature appressoria. Northern blot analysis was used to confirm the cDNA array results.


Subject(s)
Cell Proliferation , Fungal Proteins , Metabolism , Fungal Structures , Metabolism , Gene Expression Profiling , Methods , Magnaporthe , Metabolism , Oligonucleotide Array Sequence Analysis , Methods , Proteome , Metabolism
6.
China Journal of Chinese Materia Medica ; (24): 1719-1721, 2006.
Article in Chinese | WPRIM | ID: wpr-315971

ABSTRACT

<p><b>OBJECTIVE</b>To study the effects of Annao tablet (main component is beta-asarone) on S100B and NPY of cortex in chronic epilepsy rats.</p><p><b>METHOD</b>The remedy was administered orally. The effects were observed in convulsion model induced by PG, then S100B protein and NPY of cortex were determined.</p><p><b>RESULT</b>Annao tablet could depress the epileptic degree, postpone spasm latent period and reduce the wet dog sample (WDS) times. The remedy could decline S100B and NPY of cortex in chronic epilepsy rats.</p><p><b>CONCLUSION</b>Annao tablet has obvious antiepileptic effects and can reduce the nerve cell damage induced by epilepsy.</p>


Subject(s)
Animals , Female , Male , Rats , Acorus , Chemistry , Anisoles , Pharmacology , Anticonvulsants , Pharmacology , Cerebral Cortex , Metabolism , Drug Carriers , Epilepsy , Metabolism , Neuropeptide Y , Metabolism , Plants, Medicinal , Chemistry , Rats, Sprague-Dawley , S100 Proteins , Metabolism , Tablets , beta-Cyclodextrins
7.
China Journal of Chinese Materia Medica ; (24): 764-768, 2004.
Article in Chinese | WPRIM | ID: wpr-272805

ABSTRACT

<p><b>OBJECTIVE</b>To establish the method of fingerprint analysis on volatile oil in rhizome of Acorus tatarinowii by GC-MS, and to study the main characteristic components.</p><p><b>METHOD</b>The main components of 10 samples were determined by GC-MS.</p><p><b>RESULT</b>The injector temperature was 250 degrees C. The interface temperature was 230 degrees C. The column flow was 1.3 mL x min(-1). The column pressure was 80 kPa. The detector volt was 1.4 kV. The temperature rate was 3 degrees C x min(-1). And the main characteristic components were composed of the methyleugenol (2.13%), cis-methylisoeugenol (4.48%), trans-methylisoeugenol (0.82%), gamma-asarone (4.51%), beta-asarone (66.15%), alpha-asarone (6.35%). And the RSD of precision and reproducibility and stability was almost in the range of 5%.</p><p><b>CONCLUSION</b>The method is reliable, accurate and can be used for fingerprint analysis of volatile oil of Acorus tatarinowii.</p>


Subject(s)
Acorus , Chemistry , Anisoles , Eugenol , Gas Chromatography-Mass Spectrometry , Oils, Volatile , Chemistry , Plants, Medicinal , Chemistry , Rhizome , Chemistry
8.
Acta Pharmaceutica Sinica ; (12): 836-838, 2004.
Article in Chinese | WPRIM | ID: wpr-241388

ABSTRACT

<p><b>AIM</b>To study the pharmacokinetics of beta-asarone in rats.</p><p><b>METHODS</b>The concentration of beta-asarone in serum and organs were measured by HPLC after i.g. administration, the pharmacokinetics was analyzed with DAS software regarding the organs as independent system.</p><p><b>RESULTS</b>The pharmacokinetics of beta-asarone can be described as first order process of one-compartment model. In the serum, T(1/2), Tpeak and Cmax were 54 min, 12 min and 3.19 mg x L(-1), respectively. The procedure in the organs was similar to that in serum.</p><p><b>CONCLUSION</b>The absorption, distribution and elimination of beta-asarone are very rapid, and it is easy to pass through blood brain barrier. Brain is an important organ of distributing of beta-asarone.</p>


Subject(s)
Animals , Female , Male , Rats , Acorus , Chemistry , Administration, Oral , Anisoles , Pharmacokinetics , Area Under Curve , Blood-Brain Barrier , Brain , Metabolism , Chromatography, High Pressure Liquid , Methods , Half-Life , Plant Roots , Chemistry , Plants, Medicinal , Chemistry , Rats, Sprague-Dawley , Tissue Distribution
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