Spatholobus suberectus column extract inhibit estrogen receptor positive breast cancer

OBJECTIVE To investigate the inhibitory effects of Spatholobus Suberectus Column Extract(SSCE)on estrogen receptor positive(ER+)breast cancer cel MCF-7and its possible molecular mechanism.METHODS MCF-7cells were cultured without estrogen and with 17-β-estrogen(10-8mol·L-1),respectively,then treated with SSCE(0,40,80,160,320μg·m L-1).MTT assay was employed to evaluate cell viability.Flow cytometry assays were performed to underlying apoptosis and detecting cel cycle of MCF-7 cells treated with SSCE(0,80,160,320μg·mL-1).Wound healing assays was conducted to detect the migration ability.Dual luciferase reporter system was used to detect the activity of p-ERα,p-ERβpresented in intra-nuclear estrogen response element(ERE).Western blotting assay was employed to identify the expression of protein such as Bax,Bcl-2,p-ERα,p-ERβ,ERK1/2,p-ERK1/2,AKT,p-AKT,m TOR,p-m TOR,PI3K,p-PI3K.RESULTS It showed that SSCE(80,160and 320μg·mL-1)significantly decreased the viability of MCF-7.SSCE also triggered apoptosis,arrested cell cycle at G0/G1phase,inhibited migration.Dual luciferase reporter system showed that SSCE suppressed intra-nuclear p-ER activity,Western blotting analysis confirmed that SSCE did repress the expression of phosphorylated-ER alpha(p-ERα),ERK1/2,p-ERK1/2,AKT,p-AKT,pmT OR,PI3K,p-PI3K,which indicate that SSCE suppress MAPK PI3K/AKT signal pathway.CONCLUSION Our result showed that SSCE cause ER+MCF-7 cells apoptosis,G0/G1phase arresting,migration decreasing,via hypo-active of ER,suppress MAPK PI3K/AKT pathway.

Investigation on the Mode of Action of the Traditional Chinese Medical Prescription-Yiqihuoxue Formula, an Effective Extravasation Treatment for Cerebral Vascular Microemboli in ApoE-/- mice

Objective: The objective of this study was to investigate the mechanisms underlying anti-embolism and extravasational effects of traditional Chinese medical prescription YiqiHuoxue(YQHX) formula in ApoE-/-mice with cerebral vascular microemboli. Materials and Methods: An ApoE-/-mice model with microemboli was developed by infusing fluorescently labeled heterologous fibrin-rich microparticles into the internal carotid artery of ApoE -/-gene knockout male mice through the common carotid artery. Before microemboli injection, the animals were randomly divided into four groups of 10 animals, treated daily for 6 weeks by intragastric administration: The ApoE-/-control group(physiological saline, 0.2 mL/10 g/d), YQHX group(0.2 ml/10 g/d), clopidogrel group(3 mg/kg/d), and atorvastatin group(3 mg/kg/d);a further group was constituted of normal male C57 BL/6 J mice(with the same genetic background as ApoE-/-mice;normal control group;no treatment;microemboli injection). The mice in each microemboli group were divided into three subgroups, the 2-h, 24-h, and 72-h subgroups, corresponding to the time after microemboli injection. Two hours(or 24 h or 72 h) after microemboli injection, the changes in aortic intima and brain tissue were analyzed by histopathology, the amounts of fluorescent emboli being measured by fluorescence microscopy image analysis. Comparison points included the microemboli induced loss of aorta functions and pathological changes, atherosclerotic plaque, brain ultrastructure and functions, and embolus extravasation. Results: Loss of aorta functions and adverse pathological changes, atherosclerotic plaque, serious damage in brain ultrastructure and functions, and reduced thrombus elimination were obviously serious in microemboli injected ApoE-/-mice. These symptoms were significantly relieved by the YQHX pretreatment:(i) the ratio of thrombus accumulation was increased with a significant decrease in thrombus extravasation in ApoE-/-mice, while YQHX induced an increased thrombus extravasation;(ii) the degree of aortic intimal thickening and brain tissue structural disorders were significantly increased in ApoE-/-mice, but overtly inhibited in the YQHX group;(iii) YQHX restored cell viability and homeostasis in the brain;(iv) YQHX regulated the expression of pro-and anti-inflammatory cytokines in the aorta;and(v) YQHX reduced cortical nerve nuclei pyknosis, edema, liquefaction, and necrosis induced by brain hypoxia, especially in the 24 h and 72 h groups. Conclusions: These findings indicate that the protective effects of YQHX on the brain against microemboli-induced injury may be attributed to the activation of extravasation mechanisms, which are involved in the cerebrovascular injury pathway and constitutively important in the progression of ischemic stroke.

Effect of Aqueous Extracts of Several Kinds of Herbs on Human Platelet Aggregation and Expression of P-Selectin in Vitro

Objective： To study the effect of aqueous extract of several kinds of herbs on human platelet aggregation and expression of P-selectin in vitro. Methods： Blood was collected from volunteers. Effects of the prepared water extracts of herbs on platelet aggregation were monitored on a Packs-4 aggregometer. The fluorescence intensity of water extracts of Caulis Spatholobi, Flos Carthami and Rhizoma Curcumae on the expression of P-selectin in human platelets of healthy persons was measured with flow cytometry. Results： Out of several herbs investigated, Flos Carthami and Rhizoma Curcumae potently inhibited platelet aggregation after incubation with platelet-rich plasma （PRP） for 15 min. Caulis Spatholobi, Flos Carthami and Rhizoma Curcumae inhibited adenosine-5＇-diphosphate （ADP） or platelet activating factor （PAF）-induced platelet aggregation in PRP in a dose-dependent manner. In contrast to Flos Carthami and Rhizoma Curcumae, Caulis Spatholobi could not inhibit thrombin-induced platelet aggregation. Despite its inability to inhibit thrombin-induced platelet aggregation in PRP, Caulis Spatholobi had a greater anti-aggregating activity in PRP induced by ADP or PAF. Caulis Spatholobi and Flos Carthami showed significant inhibitory effects on the expression of P-selectin. Conclusions： Caulis Spatholobi, Flos Carthami and Rhizoma Curcumae have potent anti-platelet properties, and their inhibitory actions are mediated via different mechanisms. Caulis Spatholobi inhibited ADP-induced platelet aggregation but not by thrombin, indicating that its mechanism of action might be independent of the thromboxane pathway. The effect of Caulis Spatholobi and Flos Carthami were associated with suppressing the expression of P-selectin.