The effects of Ankaferd Blood Stopper on transcription factors in HUVEC and erythrocyte protein profile

Abstract

Objective: Ankaferd ® Blood Stopper (ABS) is an herbal extract that has historically been used as a hemostatic agent in traditional Turkish medicine. ABS is comprised of a standardized herbal mixture of T. vulgaris, G. glabra, V. vinifera, A. officinarum, and U. dioica. ABS's basic mechanism of action is the formation of an encapsulated protein web, which represents the focal point for vital erythrocyte masses. The hemostatic effects of ABS have been observed in vitro and in vivo. ABS was registered as a hemostatic agent for external hemorrhages and dental bleeding following phase I randomized, double-blind crossover placebo-controlled clinical research, and safety and efficacy reports. In terms of the potential use of ABS, transcription factors may be novel factors that play a role in the hemostatic and other pleiotropic effects of ABS. Materials and Methods: Hence, the present study aimed to investigate the effects of ABS on endothelium, and possible transcription factor changes in HUVEC (human umbilical vein endothelial cells) and the erythrocyte membrane profile. ABS (5 μL and 50 μL) was administered to HUVEC (in 75 cm 2 ; ~75% fullness) for 5 min and 15 min. Results: ABS caused significant increases in the level of activation of the following transcription factors; AP2, AR, CRE/ATF1, CREB, E2F1-5, E2F6, EGR, GATA, HNF-1, ISRE, Myc-Max, NF-1, NFkB, p53, PPAR, SMAD 2/3, SP1, TRE/AP1, and YY1. Following erythrocyte membrane isolation, protein complexes were undissolved, but denatured. The protein complex formed was resistant to heat and detergent. Trypsin and sonication were used in order to break this complex; the complex dissolved and erythrocyte membrane proteins were released in SDS-PAGE. Conclusion: ABS established a very fast and solid protein web, and increased the level of transcription factor activation. Therefore the cellular effects of ABS could be related to different intracellular biological pathways. (Turk J Hematol 2011; 28: 276-85)

Histopathological Alterations in the Kidney Tissue Following Topical Ankaferd Hemostat Application in a Rat Renal Injury Model

“…ABS also up-regulates GA-TA/FOG transcription system affecting erythroid functions and urotensin II. [4][5][6] Therefore, our results indicating red blood cell accumulation and siderophages following the use of ABS are compatible with the suggested 'mechanism-of-action' of ABS that ABS-induced formation of the protein network with vital erythroid aggregation covers the entire physiological hemostatic process. Further experimental search is needed to find out the molecules inside the ABS protein library leading to the ABSinduced aggregation at the renal tissue level.…”

“…Therefore, there are molecular links underlying ABSpleiotropic cellular actions acting on anti-infective, wound healing, and apoptotic processes. 1,6,7,10,13,17,[24][25][26][27][28][29][30][31][32] Since renal system is located at the crossroads of cellular neoplastic development and protein secretions, the unique effects of ABS on distinct renal cell types shall be investigated in future in vivo animal models mimicking cancer models. Our present study represent true basis for the design of such experimental animal trials.…”

“…Those proteins and the required ATP bioenergy are included in the ABS protein library. [4][5][6] Ankaferd also upregulates GATA/FOG transcription system affecting erythroid functions. Urotensin II is also an essential component of Ankaferd and represents the link between injured vascular endothelium, adhesive proteins, and active erythroid cells.…”

“…Those concepts have been developed via MALDI-TOF proteomic molecular analyses, cytometric arrays, transciption analysis, and SEM ultrastructural examinations as well as numerous investigations interacting with in vitro and in vivo research settings. 1,[3][4][5][6][7][8] ABS has many cellular effects. ABS has been shown to affect renal tubular apoptosis based on the level of hemorrhage.…”

Functional proteomic analysis of Ankaferd® Blood Stopper

“…Ankaferd represents its unique local hemostatic effect by promoting the very rapid (<1 s) formation of a protein network, which acts as an anchor for vital physiological erythrocyte aggregation, covering the classical cascade model of the clotting system without independently acting on coagulation factors and platelets [1,12]. Unique effects of critical proteins inside the ABS on critical transcription factors [14] and in vitro anti-infectious [15] and anti-cancer [16,17] effects suggested that Ankaferd may also affect pathobiological courses of tissues in addition to its unique action on hemostasis [18].…”