Objective To investigate the variation of the preparation time of Yuhong Ointment under conditions of accelerated test and long-term test;To provide the necessary data for the production and new medicine application, and establish the period of validity. Methods Referring to the current guiding principles of TCM stability test, the conditions of affecting factors in trial test and long-term test were decided:β,β'-dimethylacrylshikonin content was set as quantitative indicators, combined with the key items of stability test to evaluate the stability;its validity predictive value was deduced by using statistical methods. Results In the conditions of high temperature (30 ± 2 ℃, RH 45% ± 5%), high humidity (25 ± 2 ℃, RH 75% ± 5%), and hard light (4500 ± 500 Lx, 25 ± 2 ℃, RH 45%± 5%) for 10 days, the traits, appearance and content were in line with requirements. The validity of Yuhong Ointment under 25 ℃ conditions was 41.216 months. Conclusion Under current production conditions, the stability of Yuhong Ointment is good.

OBJECTIVETo investigate the effect of monocyte chemotactic protein-3 (MCP-3) on the expressions of intercellular adhesion molecule-1 (ICAM-1), vascular cell adhesion molecule-1 (VCAM-1), tissue factor (TF, and tissue factor pathway inhibitor (TFPI) and cell apoptosis in human umbilical vein endothelial cells (HUVECs).

METHODSCultured HUVECs were treated with MCP-3 at the optimal concentration determined previously 1 h after treatments with or without MCP-3 antibody (20 ng/ml), PI3K inhibitor, or LY-294002 (5 mmol/ml). The expressions of ICAM-1, VCAM-1, TF and TFPI were analyzed using RT-PCR and Western blot after the treatments. MCP-3 mRNA and protein expressions were detected in HUVECs exposed to 50 µg/ml ox-LDL for 24 h. The cell apoptosis and caspase-3 protein production in HUVECs treated with MCP-3 or with MCP-3 plus CCR2 antagonist for 24 h and 48 h were evaluated by flow cytometry and Western blotting.

RESULTSAt the optimal concentration of 0.3 ng/ml, MCP-3 treatment for 24 h caused significantly increased ICAM-1, VCAM-1, and TF expressions with lowered expression of TFPI in HUVECs (P<0.05), and such effects were significantly inhibited by the application of MCP-3 antibody, PI3K inhibitor, or LY-294002 (P<0.05). Ox-LDL exposure significantly increased the expression of MCP-3 in HUVECs (P<0.05). HUVECs showed a significantly increased apoptosis rate after treatment with MCP-3 or with MCP-3 plus CCR2 antagonist (P<0.05), and the apoptosis rate increased significantly as the treatment time prolonged (P<0.05); caspase-3 protein expression in the cells showed a similar pattern of alterations following the treatments.

CONCLUSIONox-LDL can induce MCP-3 expression in HUVECs. MCP-3 induces apoptosis of HUVECs and significantly affects the cellular function partially through the PI3K signaling pathway.

Apoptosis ; drug effects ; Caspase 3 ; metabolism ; Cell Adhesion ; Cells, Cultured ; Chemokine CCL7 ; pharmacology ; Chromones ; pharmacology ; Human Umbilical Vein Endothelial Cells ; cytology ; drug effects ; metabolism ; Humans ; Intercellular Adhesion Molecule-1 ; metabolism ; Lipoproteins ; metabolism ; Lipoproteins, LDL ; pharmacology ; Morpholines ; pharmacology ; Phosphatidylinositol 3-Kinases ; antagonists & inhibitors ; Receptors, CCR2 ; antagonists & inhibitors ; Signal Transduction ; Thromboplastin ; metabolism ; Vascular Cell Adhesion Molecule-1 ; metabolism

OBJECTIVETo investigate the effects of Bushen Huoxue Fang on the proliferation of rat cardiac fibroblasts and collagen production in the cells.

METHODSRat cardiac fibroblasts were isolated and cultured in DMEM containing 10% (group A) or 20% (group B) or no (group C) serum from rats treated with Bushen Huoxue Fang, with cells cultured in DMEM containing 10% FBS as the control (group D). After 72 h of cell culture, the proliferation of the fibroblasts was detected using CCK-8 kit, and collagen mRNA and protein expressions were examined using RT-PCR and Western blotting, respectively.

RESULTSCompared with that in groups C and D, the cell proliferation decreased significantly in groups A and B, and especially in the latter (P<0.05). RT-PCR demonstrated significant reductions of the mRNAs of type 1 and 3 collagens in groups A and B (P<0.05), and their protein levels were also significantly lowered (P<0.05).

CONCLUSIONBushen Huoxue Fang can effectively inhibit the proliferation of rat cardiac fibroblasts and reduced collagen type 1 and 3 productions in the cells in vitro.

Animals ; Animals, Newborn ; Cell Proliferation ; drug effects ; Cells, Cultured ; Collagen Type I ; biosynthesis ; Collagen Type III ; biosynthesis ; Drugs, Chinese Herbal ; pharmacology ; Fibroblasts ; cytology ; metabolism ; Fibrosis ; prevention & control ; Myocardium ; cytology ; metabolism ; pathology ; Rats ; Rats, Sprague-Dawley