Hyaluronidases play very important roles in a number of physiological and pathological processes such as embryogenesis, angiogenesis, inflammation, disease progression and wound healing [1 ]. Human hyaluronidase-1 (Hyal-1) is an enzyme strongly involved in the regulation of extracellular matrix by balancing the deposition and potential degradation of hyaluronic acid (HA) in the tissue [2 ]. The inhibition of Hyal-1 by specific inhibitors might be a promising target for improved wound healing, tissue regeneration, and looking at renal function also for induction of diuresis. By using surface-displayed human Hyal-1 on Escherichia coli F470 cells, HA as substrate and stains-all method for quantification of undegraded, high molecular polymer, the enzyme activity can be determined easily [3 ]. From a selection of herbal materials, traditionally used for wound healing and for urinary tract infections IC50 values were determined within a screening. Based on these results, Ononis spinosa L. roots were chosen for bioactivity guided fractionation. Two non-polar fractions of Ononis spinosa L. roots (ODB and ODG) were found to be most active following bioassay-guided fractionation with 86±3 % and 96±13 % inhibition respectively for 1mg/ml concentration. Further separation of ODB revealed 3 main components which were determined to be onogenin, sativanone and medicarpin. Relative inhibitions for 250µM concentrations were 25.3±18, 61.20 ± 20.6 and 22.4±16 respectively. The IC50 of sativanone was determined to be 140.4 µM. Rationalization of the diuretic properties of roots of Ononis spinosa L. is possible based on the results.
References
1.Heldin, P., Braz J Med Biol Res, 2003. 36(8): p. 967-73.
2.Stern, R., Pathol Biol (Paris), 2005. 53(7): p. 372-82.
3.Orlando, Z., et al., Molecules, 2015. 20(9): p. 15449-68.
Figure 1: Sativanone