In vitro and in vivo evaluation of the antiangiogenic activities of Trigonella foenum-graecum extracts

Objective: To assess the antiangiogcnic activity of fenugreek.Methods: Different fractions of fenugreek crude extracts were prepared and their antiangiogenic properties were assessed using the ex vivo rat aortic ring assay and in vivo chicken embryo chorioallantoic membrane(CAM) assay. They were investigated for their direct cytotoxic activity in the MCF7 cells using the MTT assay.Results: The ethanol extract showed 100% inhibition of blood vessel outgrowth from primary tissue explants in the rat aortic ring assay at a concentration of 100μg/mL while the other extracts did not show significant antiangiogenic activity. The ethanol extract was therefore investigated at varying concentrations and exhibited a significant dose dependent effect. The CAM assay coincided with the results of the aortic ring assay as ethanol extract showed a significant inhibition of formation of new blood vessels. The extracts only showed anti-proliferative activity at the highest concentration of 400μg/mL towards MCF7 breast cancer cell lines in the MTT assay.Conclusions: Findings of the both assays confirmed that the ethanol extract inhibited vascularization significantly. Further studies on the ethanol extract would be beneficial in isolating the active ingredient responsible for the inhibition.

Anti-proliferative and anti-angiogenic activities of ion-channel modulators:In-ovo,in-vitro and in-vivo study

Objective:Angiogenesis is the development of new blood vessels.The ion channels on endothelium play a vital action in cell proliferation and so in the related angiogenesis.We aimed to investigate the anti-angiogenic effects of Mefloquine(Cl-channel blocker) and4-Aminopyridine(K+ channel blocker).Methods:The anti-angiogenic activities of Mefloquine and 4-Aminopyridine(4-AP)were investigated by in-vivo(sponge implantation method),in-vitro(aortic ring assay)and in-ovo(CAM,Chick Chorioallantoic membrane) methods.The standard antiangiogenic drug used was Bevacizumab.Results:In the CAM assay,both the ion channel blockers exhibited noticeable antiangiogenic activity at the concentrations of 10-5M and 10-4M where they significantly exhibited ant proliferative activity by inhibiting the new blood vessel formation.For the further confirmation anti-angiogenic activity was evaluated in vitro and in vivo.In Rat aortic ring assay reduction in the area of sprouts were observed with 40 m M of 4-AP and7 m M of Mefloquine.A significant reduction in weight of sponges,number of blood vessels formed and hemoglobin content were observed at 4.2 mg/kg of 4-AP and 20 mg/kg and 30 mg/kg of Mefloquine.Conclusions:These scientific findings indicate the use of Mefloquine and 4-Aminopyridine in pathological situations involving excessive angiogenesis.Negative regulation of cell volume,cell migration and proliferation of blood vessels may be the underlying molecular mechanisms.

Molecular mechanism of Chuanxiong Rhizoma in treating coronary artery diseases

Objective:Most of the studies on the herb Chuanxiong Rhizoma(CR)have focused on the L-arginine-nitric oxide(NO)pathway,but the nitrate-nitrite-NO(NO_(3)^(-)–NO_(2)^(-)–NO)pathway was rarely investigated.Therefore,the aim of this study was to evaluate the effects and mechanisms of action of CR in coronary artery disease(CAD).Methods:The NO_(3)^(-),NO_(2)^(-)and NO levels were examined in the NO_(3)^(-)–NO_(2)^(-)–NO pathway.High-performance ion chromatography was used to quantify NO_(3)^(-)and NO_(2)^(-)levels.Then,NO was quantified using a multifunctional enzyme marker with a fluorescent probe.The tension of aortic rings was measured using a multi myograph system.Results:High content of NO_(3)^(-)and low content of NO_(2)^(-)was found in CR,and which could potently convert NO_(3)^(-)to NO_(2)^(-)in the presence of endogenous reductase enzyme.Incubating human coronary artery endothelial cells(HCAECs)with CR-containing serum showed that CR significantly decreased the NO_(3)^(-)content and increased the levels of NO_(2)^(-)and NO in the cells under hypoxic conditions.In addition,CR significantly relaxed isolated aortic rings when the L-arginine–NO pathway was blocked.The optimal concentration of CR for relaxation was 200 mg/m L.Conclusion:CR supplements large amounts of NO in cells and vessels to achieve relaxation via the NO_(3)^(-)–NO_(2)^(-)–NO pathway,thereby making up for the deficiency caused by the lack of NO after the L-arginine-NO pathway is suppressed.This study also supports the potential use of a traditional Chinese herb for future drug development.