OBJECTIVEThe study was aimed to investigate the inhibitory effect and mechanism of astragaloside IV (ASI) on the activation of microglial cells.

METHODAfter pre-incubated with ASI for 2 h, microglial cells BV-2 were stimulated with interferon-γ (IFN-γ) for 1. 5 h and 24 h, respectively. Secretion of nitric oxide (NO) in the medium was measured by Griess method. Production of tumor necrosis factor alpha (TNF-α) was detected by ELISA approach. Cellular gene expressions of CD11b, TNF-α, interleukin 1β (IL-1β) and induced nitric oxide synthase (iNOS) were examined by quantitative-PCR analysis. Total and phosphorylation of STAT1, IκB and NF-κB was analyzed by Western blot method.

RESULTASI could significantly inhibit the increased secretion of TNF-α and NO from BV-2 cells upon IFN-γ stimulation (P < 0.001). Further study showed that ASI significantly down-regulated gene expression of IL-1β and TNF-α (P < 0.01, P < 0.05) and exhibited a trend to reduce that of iNOS. IFN-γ and ASI have no obvious effect on gene expression of CD11b. Moreover, ASI inhibited the phosphorylation of STAT1, IκB and NF-κB elicited by IFN-γ stimulation.

CONCLUSIONASI could restrain microglial activation through interfering STAT1/IκB/NF-κB signaling pathway, reducing gene expres- sion of IL-1β and TNF-α, and thus inhibiting the production of proinflammatory mediators such as NO and TNF-α.

Animals ; Astragalus Plant ; chemistry ; Drugs, Chinese Herbal ; pharmacology ; I-kappa B Proteins ; genetics ; metabolism ; Interferon-gamma ; genetics ; metabolism ; Mice ; NF-kappa B ; genetics ; metabolism ; Nitric Oxide ; metabolism ; Nitric Oxide Synthase Type II ; genetics ; metabolism ; STAT1 Transcription Factor ; genetics ; metabolism ; Saponins ; pharmacology ; Signal Transduction ; drug effects ; Triterpenes ; pharmacology

OBJECTIVETo observe the effect of pure total flavonoids from Citrus (PTFC) on the hepatic fatty degeneration, inflammation, oxidative stress and SIRT1/PGC-1alpha expressions in mice with non-alcohol steatohepatitis (NASH), and discuss the action mechanism of PTFC on NASH.

METHODMice were given high-fat diet for 16 weeks to induce the NASH model. Since the seventh week after the model establishment, the mice were intervened with 100, 50 and 25 mg x kg(-1) x d(-1) PTFC for 10 weeks. The pathologic changes in hepatic tissues were observed with HE staining. The contents of TG, CHOL in hepatic tissue, as well as the levels of AST, ALT in serum were detected by using the biochemical process. The expression of SIRT1, PGC-1alpha and MnSOD mRNA in hepatic tissues were detected with Real-time PCR assay. SIRT1, PGC-1alpha protein and 8-OHdG expressions were determined with the immunohistochemical method. The SOD level in hepatic tissues was tested by the xanthine oxidase method. The MDA content in hepatic tissues was examined by the thiobarbituric acid method.

RESULTThe contents of TG, CHOL, NAFLD activity scores and ALT level in serum in hepatic tissues of mice in the model induced by fat-rich diet were obviously higher than that of the normal group (P < 0.010. The SIRT1, PGC-1alpha, MnSOD mRNA and protein expression in hepatic tissues were significantly lower than that of the normal group (P < 0.01). The expression of 8-OHdG and the content of MDA in hepatic tissues were obviously higher than that of the normal group (P < 0.01). After the intervention with different doses of PTFC, the NAFLD activity scores, the content of TG and the level of AST in serum were notably lower than that of the normal group (P < 0.01, P < 0.05); whereas the SIRT1, PGC-1alpha, MnSOD mRNA and protein expression were obviously higher than that of the normal group (P < 0.01, P < 0.05), with the significant decrease in the expression of 8-OHdG and the content of MDA (P < 0.01).

CONCLUSIONOxidative stress/lipid peroxidation enhancement in in NASH mice induced by high-fat diet may be related to the changes in SIRT1/PGC-1alpha signal transduction pathway. PTFC could enhance the anti-oxidant capacity in liver, relieve the damage of reactive oxygen during the fatty acid metabolic process, and prevent NASH from the occurrence and development by regulating the SIRT1/PGC-1alpha signal pathway.

Animals ; Citrus ; chemistry ; Fatty Liver ; drug therapy ; genetics ; metabolism ; Flavonoids ; chemistry ; pharmacology ; Inflammation ; drug therapy ; genetics ; metabolism ; Liver ; drug effects ; metabolism ; Male ; Mice ; Mice, Inbred C57BL ; Non-alcoholic Fatty Liver Disease ; Oxidative Stress ; drug effects ; genetics ; Peroxisome Proliferator-Activated Receptor Gamma Coactivator 1-alpha ; Sirtuin 1 ; genetics ; metabolism ; Transcription Factors ; genetics ; metabolism

This study aimed to investigate the involved immunologic mechanism of pure total flavonoids from Citrus (PTFC) on the development of non-alcoholic steatohepatitis (NASH). C57BL/6 mice were fed with high .fat diet for 16 weeks to induce the NASH model, and from the 7th week three dosages (25, 50 and 100 mg x kg(-1) x d(-1)) of PTFC were administrated intragastric for 10 weeks respectively. Serum TG, CHOL, ALT, AST were determined by biochemical assay, histopathological changes of the liver tissue were observed by HE staining, expression of RORyt and Foxp3 mRNA of the liver tissue was detected by Real-time PCR, and serum IL-17, IL-6, IL-10 and IL-4 were determined by.Cytometric Beads Array. As a result, we find that after the administration of PTFC, the in- flammation of the liver tissue of NASH mice was attenuated, liver function was improved, and the expression of RORgammat mRNA was higher in the liver tissue while which was lower of Foxp3 mRNA, the level of proinflammatory cytokines IL-17 and IL-6 decreased and the level of suppressive cytokines IL-10 and IL-4 increased. These data show that PTFC protects the development of NASH through regulating the Th17/Treg balance and attenuating inflammation.
Animals ; Citrus ; chemistry ; Cytokines ; blood ; Flavonoids ; pharmacology ; Male ; Mice ; Mice, Inbred C57BL ; Non-alcoholic Fatty Liver Disease ; immunology ; prevention & control ; T-Lymphocytes, Regulatory ; drug effects ; Th17 Cells ; drug effects

To study the preventive effect of Grifola frondosa on nonalcoholic steatohepatitis (NASH). The rat model of NASH was established by feeding high-fat diets for 12 weeks and intervened with 0.5 g · kg(-1) · d(-1) and 1.0 g · kg(-1) · d(-1) of C. frondosa powder suspensions. The degrees of hepatocyte fatty degeneration and inflammation were observed under the optical microscope with routine HE staining. The NAFLD activity scores (NAS) were calculated. Serum ALT, AST and hepatic TG and CHOL were tested by the biochemical method. The hepatic MDA was examined by thiobarbituric acid method. The hepatic SOD was tested by the xanthine oxidase test. The hepatic GSH-PX activity was determined by the dithio-nitrobenzoic acid method. Hepatic TNF-α and IL-6 were detected by the enzyme-linked immunosorbent assay (ELISA). The NASH model group induced by high-fat diets showed higher hepatic NAS, ser- um ALT, AST, CHOL and hepatic TG, CHOL, MDA, TNF-α, IL-6 (P < 0.01 or P < 0.05) and lower serum TG and hepatic SOD, GSH-PX (P < 0.01, P < 0.05) than the normal control group. After being intervened with different doses of G. frondosa, the NASH group revealed significantly lower hepatic NAS, serum ALT and hepatic TG, CHOL, MDA, TNF-α and IL-6 (P < 0.05) and higher hepatic SOD, GSH-PX (P < 0.05) than the model group. G. frondosa may prevent the further development of NASH by improving the disorder of lipid metabolism in rats with NASH induced by high-fat diets, relieving the level of oxidative stress and reducing the generation of inflammatory cytokines.
Animals ; Drugs, Chinese Herbal ; administration & dosage ; Grifola ; chemistry ; Humans ; Interleukin-6 ; metabolism ; Liver ; drug effects ; metabolism ; Male ; Non-alcoholic Fatty Liver Disease ; drug therapy ; metabolism ; Oxidative Stress ; drug effects ; Rats ; Rats, Sprague-Dawley ; Tumor Necrosis Factor-alpha ; metabolism

Our work focuses on the quality control and structural identification of Photocyanine as a cancer therapeutic photosensitizer. Photocyanine is a mixture which contains four ZnPcS2P2 type substituted Phthalocyanine isomers. In order to obtain the single component from Photocyanine, the mixture of four isomers possessing the similar structures and chemical property had been isolated and purified. An HPLC method with a mixture of methanol-acetonitrile-ion-pair buffer as the mobile phase was applied to isolate the four isomers by means of a semi-preparative C18 column. To remove the salts which were mixed in the preparative product, a SPE C18 column was used to separate the salts by elution with water and then the marker component was eluted by methanol. Subsequently, a column of Sephadex LH-20 gel was applied to elute the crudes with methanol to desalination. The purity of the isolated compound was measured by TLC and four different isomers of phthalocyanine were obtained. The chemical structures of them were elucidated by 1H NMR spectra, gCOSY and NOE1D. An HPLC-DAD method was developed for simultaneously determination of four major isomers in Photocyanine with a C18 column (Grace Smart, 150 mm x 4.6 mm ID, 5 microm). The separation was carried out with a gradient program at a flow rate of 1.0 mL x min(-1). The mobile phase was a mixture of acetonitrile and ion-pair buffer (0.01 mol x L(-1) hexadecyl trimethyl ammonium bromide and 0.01 mol x L(-1) potassium dihydrogen phosphate, adjusted the pH value to 6.8 with potassium hydroxide solution). The resolution values of four isomers were 2.5, 1.20, 1.33, and 1.8. Linear regression analysis for four compounds was performed by the external standard method. Four constituents were linear in the concentration range of 0.005 to 10 microg. The values of relative standard deviation (RSD) of intra-day were 0.12%, 0.66%, 0.99%, and 1.21%, respectively. The limits of detection for four compounds were 15 ng, 20 ng, 12 ng, and 25 ng, respectively. This method was simple, accurate and reproducible. The developed method can be successfully applied to analyze isomers in Photocyanine.
Antineoplastic Agents ; analysis ; chemistry ; Chromatography, High Pressure Liquid ; methods ; Indoles ; analysis ; chemistry ; Isomerism ; Molecular Structure ; Organometallic Compounds ; analysis ; chemistry ; Photochemotherapy ; Photosensitizing Agents ; analysis ; chemistry ; Quality Control

The DNA segment of the human acyl coenzyme A: cholesterol acyltransferasel (ACAT1) gene P7 promoter was amplified by PCR from human monocytic leukemia cell line (THP-1) and cloned to TA vector, then the positive clone was confirmed by restriction enzymes and sequencing. The targeted segment was subcloned to Firefly luciferase report vector pGL3-Enhancer. The recombinant plasmid pGL3E-P7 was transfected transiently into THP-1, then the expression of luciferase could be detected in THP-1 by pGL3E-P7 transfection. We successfully constructed luciferase reporter vector containing P7 promoter of the human ACAT1 gene, and established a new means to study the transcriptional regulation mechanisms of ACAT1 during atherosclerosis.
Cell Line, Tumor ; Chromosomes, Human, Pair 7 ; genetics ; Gene Expression Regulation ; Genes, Reporter ; genetics ; Genetic Vectors ; genetics ; Humans ; Leukemia, Monocytic, Acute ; pathology ; Luciferases, Firefly ; genetics ; metabolism ; RNA, Messenger ; genetics ; metabolism ; Recombinant Proteins ; genetics ; metabolism ; Sterol O-Acyltransferase ; genetics ; metabolism ; Transfection

Adolescent ; Adult ; Aged ; Aged, 80 and over ; Carcinoma, Renal Cell ; genetics ; metabolism ; Female ; Gene Expression Regulation, Neoplastic ; Humans ; Kidney Neoplasms ; genetics ; metabolism ; Male ; Matrix Metalloproteinase 2 ; genetics ; metabolism ; Middle Aged ; Transforming Growth Factor beta1 ; genetics ; metabolism ; Tumor Cells, Cultured ; Young Adult

OBJECTIVE: To explore the effective intracranial surgical method for hemifacial spasm which can be operated more safely and easily. METHOD: Twenty-five cases of hemifacial spasm underwent intracranial facial nerve root combing scraping by improved retrosigmoid approach. The compressing vessels were found in 10 cases and could be removed easily. Both microvascular decompression and facial nerve root combing scraping were performed to reduce recurrence. The other cases with perforating artery or without any compressing vessels underwent the facial nerve root combing scraping and cutting off partially for about 1/6 to approximately 1/5 neural fibers. RESULT: Hemifacial spasms were obliterated in 22 cases (88%) after operation, obviously relieved in 2 cases (8%), invalid in 1 case (4%). Varied facial nerve paralysis was observed in 25 patients, but no hearing disturbance and other serious complication occurred. Follow-up for 2 to approximately 8 years showed there were recurrences in 2 cases (8%) and the facial nerve paralysis of 25 cases recovered in 1 to approximately 6 months after operation. CONCLUSION The facial nerve root combing scraping is an effective and safe microsurgical procedure which has wide-ranged indication and less complication. It is very important to select suitable treatment for the patient individually during the operation to improve the efficacy and lower the risk.
Adult ; Aged ; Decompression, Surgical ; Facial Nerve ; surgery ; Facial Paralysis ; surgery ; Female ; Hemifacial Spasm ; surgery ; Humans ; Male ; Microsurgery ; methods ; Middle Aged ; Young Adult

Objective To investigate the effects of monosodium tetrahexose ganglioside sodium (GM-1) combined with edaravone in the treatment of acute cerebral infarction (ACI) in patients with serum homocysteine (Hcy), high sensitivity C-reactive protein (Hs-CRP). Methods 66 patients with ACI were randomly divided into observation group and control group, 33 cases were treated with conventional neurology. The control group was treated with edaravone. The observation group was treated with edaravone and GM-1. (NIHSS) score, serum hs-CRP and Hcy levels were compared between the two groups. Results The total effective rate of the observation group was 90.91%, which was significantly higher than that of the control group (P<0.05). After 21 days of treatment, the NIHSS score, serum Hcy and hs-CRP in the observation group were significantly lower than those in the control group (P<0.05 ). Conclusion GM-1 combined with edaravone is effective in the treatment of ACI, which can promote the rehabilitation of neurological function and living activity. Its mechanism may be related to its anti-inflammatory response, reduced Hcy level and protection of brain cell injury.

OBJECTIVETo provide experimental data for the quality control of processed Paeonia lactiflora, a Chinese herbal medicine.

METHODTraditional processing of P. lactiflora was simulated, content of paeoniflorin and water extracts among different preparations were assayed by HPLC; The quantitative correlations among different processing conditions were analyzed, the effects of processing parameters on the contents of paeoniflorin and water extracts were assayed a nanalysed.

RESULT AND CONCLUSIONThe controlled processing parameters were correlated with covariables which showed that processing procedures was controllable, and the heating temperature was a factor impacting the content of paeoniflorin.

Benzoates ; analysis ; Bridged-Ring Compounds ; analysis ; Glucosides ; analysis ; Hot Temperature ; Monoterpenes ; Paeonia ; chemistry ; Plants, Medicinal ; chemistry ; Quality Control ; Technology, Pharmaceutical ; methods ; Temperature