Bu Yang Huan Wu decoction prevents reperfusion injury following ischemic stroke in rats via inhibition of HIF-1 α, VEGF and promotion ß-ENaC expression.

ETHNOPHARMACOLOGICAL RELEVANCE: Bu Yang Huan Wu Decoction (BYHW) is a famous traditional Chinese medicine (TCM) formula used in China for the treatment of cerebral ischemic stroke. But the protective effects and underlining mechanisms of BYHW remain unclear. AIM OF THE STUDY: This study was designed to investigate the protective effects and underlining signaling mechanisms of BYHW on brain tissues in a rat model of cerebral ischemic reperfusion (I/R) injury. MATERIALS AND METHODS: Liquid chromatography was used to verify the composition of BYHW. The cerebral edema and infarct volume were measured by magnetic resonance imaging (MRI). The morphology and ultrastructure of ischemic penumbra brain tissues were observed by hematoxylin-eosin (HE) and transmission electron microscopy (TEM). The expression levels of HIF-1 α, VEGF and ß-ENaC were tested using immunohistochemistry technique, western blot and quantitative PCR analysis, respectively. RESULTS: Administration of BYHW significantly decreased cerebral edema, rat neurological function scores, reduced brain infarct volume. At the same time, BYHW had protective effect on the blood-brain barrier (BBB), which improved the morphology and ultrastructure of ischemic penumbra brain tissues. BYHW treatment significantly decreased the protein and mRNA levels of HIF-1 α and VEGF compared with the model treatment. In addition, BYHW treatment significantly up-regulated the protein and mRNA levels of ß-ENaC. CONCLUSIONS: BYHW protected against cerebral I/R injury in MCAO rats through inhibiting the activation of the HIF-1 α /VEGF pathway and stabilizing ion channel of ß-ENaC in brain, indicating that BYHW shows potential for stroke treatment in acute stage.

Effect of Hua Yu Xiao Zheng decoction on the expression levels of vascular endothelial growth factor and angiopoietin-2 in rats with endometriosis.

The aims of the present study were to investigate the effects of a traditional Chinese medicine, Hua Yu Xiao Zheng (HYXZ) decoction, on surgically induced endometriosis in a rat model and to determine the possible underlying regulatory mechanisms. A total of 108 female Sprague-Dawley rats were divided into the control group (n=12) and endometriosis group (EM group; n=96), in which endometriosis was surgically induced in model rats by autotransplantation of endometrial tissues and 72 rats survived. After 3 weeks, the EM model rats were randomly divided into four subgroups (n=18), including the untreated model group, and three groups administered 7, 14 or 21 g/kg HYXZ decoction. Following 28 days of treatment, the associated proteins and genes of ectopic endometrial tissues were analyzed using immunohistochemistry, western blotting and quantitative polymerase chain reaction to investigate the underlying mechanisms. Compared with the model group, the size of the endometriotic implants decreased significantly in the HYXZ-treated groups. Furthermore, the expression levels of vascular endothelial growth factor (VEGF) and angiopoietin-2 (Ang-2) were significantly decreased in HYXZ-treated groups compared with the model group. These results indicate that HYXZ affected the inhibition of angiogenesis and decreased the endometriotic implant volumes and histopathological scores. The effectiveness of HYXZ may be partially attributed to the decrease of VEGF and Ang-2 expression levels in the ectopic endometrium.

Pure total flavonoids from Citrus paradisi Macfadyen act synergistically with arsenic trioxide in inducing apoptosis of Kasumi-1 leukemia cells in vitro.

To investigate the potential effects of pure total flavonoid compounds (PTFCs) from Citrus paradisi Macfadyen separately or combined with arsenic trioxide on the proliferation of human myeloid leukemia cells and the mechanisms underlying the action of PTFCs. The effects of PTFCs separately or combined with arsenic trioxide on the proliferation and apoptosis of leukemia cells were determined by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT), fluorescence microscopy, and flow cytometry. Their effects on the expression levels of apoptosis-related regulators were determined by Western blot assay. PTFCs combined with arsenic trioxide significantly inhibited the growth of Kasumi-1 cells, and apoptosis was confirmed by flow cytometry analysis. Hoechst 33258 staining showed more significant morphological changes and more apoptosis following the combined treatment. Western blots showed changes in the expression of genes for poly ADP-ribose polymerase (PARP), caspase 3/9, and P65. The results indicated that PTFCs separately or combined with arsenic trioxide inhibited proliferation of leukemia cells in vitro and induced their apoptosis by modulating the expression of apoptosis-related regulator genes.

[UPLC-MS/MS determination of content of three iridoids of xingnaojing oral preparation in rat brains and study on their brain pharmacokinetics].

To establish a UPLC-MS/MS method for the simultaneous determination of geniposide, genipin 1-O-beta-D-gentiobioside and geniposidic acid in rat brains and study the brain pharmacokinetics of the three iridoid glycosides in stroke rat after the oral administration of Xingnaojing. In this experiment, brain samples were precipitated with protein for twice. Acquity BEH C18 column was adopted, with acetonitrile-0.1% formic acid-water as the mobile phase for gradient elution. ESI source was adopted for mass spectra; multiple reaction monitoring (MRM) was conducted to detect negative ions. The time for sample analysis was 3.5 min. the results showed good linear relations among the three iridoid glycosides, with the extraction recovery between 99.6% and 114.3%, good intra- and inter-day precisions and accuracies and stability in line with the requirements. The t1/2 and MRT in the three components were similar in brains of stroke rats. Geniposide and genipin 1-O-beta-D-gentiobioside showed double peaks; where as geniposidic acid showed a single peak. In conclusion, the method is so specific, sensitive, accurate and reliable that it can be used to study the brain pharmacokinetics of Xingnaojing oral preparation.

Delisheng Injection (), a Chinese medicinal compound, enhanced the effect of cis-platinum on lung carcinoma cell line PGCL3.

OBJECTIVE: To investigate the effect of Delisheng Injection (, DLS), a Chinese medicinal compound, DLS combined with cis-platinum (DDP), an active agent used in lung cancer chemotherapy, on a human highly metastatic giant lung carcinoma cell line PGCL3. METHODS: The suspended PGCL3 cells at 10(5) /mL cultured in 96-well tissue culture plates were divided into 4 groups: DLS treatment group (2 µL/mL, 5 µL/mL, 10 µL/mL, 25 µL/mL), DDP treatment group (1 µg/mL, 2 µg/mL, 5 µg/mL, 15 µg/mL), combined DLS with DDP treatment group (DLS:DDP 2 µL/mL:1 µg/mL, 5 µL/mL:2 µg/mL, 10 µL/mL:5 µg/mL, 25 µL/mL:15 µg/mL) and a control group. The cytotoxicity of DLS with different concentrations (2 µL/mL, 5 µL/mL, 10 µL/mL, 25 µL/mL) on PGCL3 cells was determined by 3-(4,5)-dimethylthiahiazo(-z-y1)-3,5-di-phenytetrazoliumromide (MTT) assay. Effect of DLS on adhesion of PGCL-3 cells was tested by cell-matrigel adhesion assay. Chemotactic movement model of transwell camerula was used to determine the effect of DLS on invasion and migration of PGCL-3 cells. RESULTS: Compared with the control group, DLS (2 µL/mL, 5 µL/mL, 10 µL/mL, 25 µL/mL) could significantly decrease cell proliferation, adhesion, invasion and migration abilities (P <0.05). Cell adhesion, invasion and migration abilities were significantly decreased after combination treatment of DLS:DDP (2 µL/mL:1 µg/mL, 5 µL/mL:2 µg/mL, 10 µL/mL:5 µg/mL, 25 µL/mL:15 µg/mL) compared with DDP single-agent treatment (1 µg/mL, 2 µg/mL, 5 µg/mL, 15 µg/mL, P<0.05), respectively. CONCLUSIONS: DLS single-agent has a satisfying inhibition effect in PGCL3 cell line and DLS might enhance the inhibition effect of DDP on cancer metastasis. Our research provided a experimental basis about the treatment on highly metastatic lung caner.

Catalytic spectrophotometric determination of ascorbic acid in tea drink with 1,5-bis(p-hydroxybenzaldene)thiocarbohydrazone as the substrate for horseradish peroxidase.

1,5-bis(p-Hydroxybenzaldene)thiocarbohydrazone (BHBTZ) had been synthesized and used as a new kind of substrate for horseradish peroxidase (HRP). The experiment results showed that ascorbic acid (AsA) had strongly inhibiting effect on the BHBTZ-H(2)O(2)-HRP reaction system, so AsA had been spectrophotomericly determined as the inhibitor by enzymatic inhibition method. Under experimental conditions, the linear relationship between DeltaA(386) and AsA concentration was in the range of 0.82-18.0 microg ml(-1) with a detection limit of 0.245 microg ml(-1). The effect of interferences on AsA determination was investigated. The proposed method was successfully applied to the determination of AsA in tea drink.