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Article in Chinese | WPRIM | ID: wpr-905924


Objective:To observe and compare the protective effects of Tongqiao Huoxue decoction (TQHX) prepared by three methods against cerebral ischemia-reperfusion injury (CIRI), and to explore its mechanism through the glutamate (Glu) metabolic pathway in astrocytes. Method:The male SD rats of SPF grade were subjected to CIRI model induction by the modified middle cerebral artery occlusion method. The model rats were randomly divided into a model group, a sham operation group, and water-decocted, wine-decocted, and alcohol-extracted TQHX (6.3 g·kg<sup>-1</sup>·d<sup>-1</sup>) groups. The rats were treated correspondingly for 7 days. Those in the sham operation group and the model group were treated with an equal volume of normal saline by gavage. After the final treatment, the neurological function of rats was assessed by the modified neurological severity score (mNSS). Hematoxylin-eosin (HE) staining was used to observe the morphological changes of ischemic brain tissues in rats. High-performance liquid chromatography (HPLC) was used to detect glutamate (Glu) in ischemic brain tissues. The expression of glutamate transporter-1 (GLT-1) and glial fibrillary acidic protein (GFAP) and co-expression of glutamine synthetase (GS) and GFAP in ischemic brain tissues were detected by immunofluorescence assay. Western blot was used to detect the protein expression of GFAP, GLT-1, and GS. Result:Compared with the sham operation group, the model group showed increased mNSS (<italic>P</italic><0.01), large necrosis of cerebral cortex in ischemic brain tissues with disordered cell arrangement, obscure boundary, intracellular edema, and inflammatory infiltration, elevated Glu in ischemic brain tissues (<italic>P</italic><0.01), declining GLT-1-GFAP co-expression and GS-GFAP co-expression (<italic>P</italic><0.01), up-regulated expression of GFAP protein, and reduced protein expression of GLT-1 and GS(<italic>P<</italic>0.05,<italic>P<</italic>0.01). Compared with the model group, the TQHX groups showed decreased mNSS (<italic>P<</italic>0.01), relieved injury in the cerebral cortex and hippocampal nerve cells in ischemic brain tissues, reduced Glu expression(<italic>P<</italic>0.05,<italic>P<</italic>0.01), elevated co-expression of GLT-1 and GFAP (<italic>P<</italic>0.05,<italic>P<</italic>0.01), and up-regulated protein expression of GFAP and GLT-1(<italic>P<</italic>0.05,<italic>P<</italic>0.01). The co-expression of GS and GFAP (<italic>P<</italic>0.05,<italic>P<</italic>0.01)and the expression of GS (<italic>P<</italic>0.01)were increased in the wine-decocted and alcohol-extracted TQHX groups. Compared with the water-decocted TQHX group, the alcohol-extracted group showed increased GLT-1-GFAP and GS-GFAP co-expression(<italic>P<</italic>0.05); the wine-decocted and alcohol-extracted TQHX groups exhibited elevated GS protein expression (<italic>P<</italic>0.05); the alcohol-extracted TQHX group displayed declining Glu content (<italic>P</italic><0.01) and increased protein expression of GFAP and GLT-1 (<italic>P<</italic>0.05, <italic>P<</italic>0.01). Compared with the wine-decocted TQHX group, the alcohol-extracted TQHX group showed increased protein expression of GFAP and GLT-1(<italic>P<</italic>0.05,<italic>P<</italic>0.01). Conclusion:TQHX prepared by three methods can improve neurological deficits in CIRI rats. The effect is presumedly achieved by promoting the further activation of astrocytes, increasing the expression of GLT-1 and GS, promoting the clearance of Glu accumulated in the synaptic cleft by astrocytes through the Glu-glutamine (Gln) circulation, and reducing the excitotoxicity of Glu. The alcohol-extracted TQHX group was superior to the water-decocted and wine-decocted TQHX groups in reducing the content of Glu in ischemic brain tissues, promoting the activation of astrocytes, and enhancing the protein expression of GLT-1 and GS.

Basic & Clinical Medicine ; (12): 1712-1719, 2017.
Article in Chinese | WPRIM | ID: wpr-669128


Objective To investigate the inducing effects and related pathways of selenium dioxide ( SeO2 ) on the apoptosis in 2 human cervical carcinoma cell lines of high risk HPV subtypes .Methods HeLa (HPV-18-positive) and Caski (HPV-16-positive) cells were incubated with different concentrations of SeO 2 for 24 h respectively.Mor-phological changes of HeLa and Caski cells were observed under inverted optical microscope ;cell proliferation and activity were examined by MTT assay;flow cytometry was employed to detect the cell apoptosis;the expressions of apoptosis-related proteins caspase-3 and p53 in cervical carcinoma cell lines were determined by Western blot analysis;the effects of SeO 2 on apoptosis-related miRNA LET-7a expression was detected by stem-loop reverse tran-scription-polymerase chain reaction (RT-PCR).Results Cell morphology was obviously changed in vitro.Cells be-came rounded and shrunken .SeO2 markedly inhibited cell proliferation and viability in a dose-dependent manner in both cell lines; In HeLa cells the inhibitory effects induced by 7.5-30 μmol/L of SeO 2 were significant ( P<0.05);The inhibitory effects on Caski were statistical significant (P<0.05) even with low concentrations of SeO 2. The apoptosis induced by SeO 2 increased dose-dependently in cervical carcinoma cell lines , which were higher in Caski.SeO2 up-regulated the apoptosis-related proteins in cervical carcinoma cell lines .The expressions of caspase-3 and p53 in HeLa cells both significantly increased as compared with control group (P<0.05), and peaked at the concentration of 7.5 μmol/L.Treated with higher concentrations ( 7.5-30 μmol/L ) of SeO 2 , the expression on Caski cells increased significantly ( P<0.05 ) .SeO 2 induced of expression of apoptosis-related miRNA LET-7a both in HeLa cells and Caski cells with statistical meanings ( P<0.05 );the effects reached its peak at the concentration of 7.5 μmol/L bothly.Conclusions SeO2 shows anti-tumor properties via apoptosis pathway by up-regulating the expressions of apoptosis-related proteins caspase-3, the mechanisms of can be potentially explained by p 53 and LET-7a in cervical cancer cell lines.The apoptosis-inducing effect of SeO2 is more sensitive in HPV16+cell line compared with HPV18+cell line.

Chinese Pharmaceutical Journal ; (24): 1370-1373, 2012.
Article in Chinese | WPRIM | ID: wpr-860629


OBJECTIVE: To study the effect of BSO administration on the multidrug resistance (MDR) transformation of human breast cancer cell line in vitro. METHODS: The intracellular GSH content was measured by glutathione reductase recycling assay; the 50% inhibitory concentration (IC50) of ADM was measured by MTT assay, then the multidrug resistance property and multidrug resistance reversal times were determined; the apoptosis rate and cell cycle of MCF-7/ADM cell both before and after BSO administration were measured by flow cytometry. RESULTS: The level of GSH in MCF-7/ADM cells was significantly higher than that in MCF-7 cells; the BSO treatment exerted significant inhibitory effect on the synthesis of GSH in MCF-7/ADM while slight effect on MCF-7 cell; The IC50 of ADM in MCF-7/ADM cells was decreased by BSO of certain doses, the multidrug resistance reversal times by BSO at concentrations of 50, 100, 200, 400, and 800 μmol · L-1 were 1.25, 2.02, 8.42, 12.65, and 9.71 respectively, but BSO had no such effect on MCF-7 cell. The value of apoptosis rate and the cells accumulated in Go/GI phase increased significantly when BSO and ADM were simultaneously used compared with using ADM merely. CONCLUSION: BSO can potentially reverse the MDR of MCF-7/ADM cells by inhibition of GSH synthesis in vitro, and the effect may be achieved by inducing apoptotic cell death. Copyright 2012 by the Chinese Pharmaceutical Association.