Donkey milk and donkey milk kefir exhibit antiproliferative, antimutagenic and antibacterial effects. We investigated the effects of donkey milk and donkey milk kefir on oxidative stress, apoptosis and proliferation in Ehrlich ascites carcinoma (EAC) in mice. Thirty-four adult male Swiss albino mice were divided into four groups as follows: group 1, administered 0.5 ml water; group 2, administered 0.5 ml water + EAC cells; group 3, administered 0.5 ml donkey milk + EAC cells; group 4, administered 0.5 ml donkey milk kefir + EAC cells. We introduced 2.5 x 10⁶ EAC cells into each animal by subcutaneous injection. Tap water, donkey milk and donkey milk kefir were administered by gavage for 10 days. Animals were sacrificed on day 11. After measuring the short and long diameters of the tumors, tissues were processed for histology. To determine oxidative stress, cell death and proliferation iNOS and eNOS, active caspase-3 and proliferating cell nuclear antigen were assessed using immunohistochemistry. A TUNEL assay also was used to detect apoptosis. Tumor volume decreased in the donkey milk kefir group compared to the control and donkey milk groups. Tumor volume increased in the donkey milk group compared to the control group. Proliferating cell nuclear antigen levels were higher in the donkey milk kefir group compared to the control and donkey milk groups. The number of apoptotic cells was less in the donkey milk group, compared to the control, whereas it was highest in the donkey milk kefir group. Donkey milk administration increased eNOS levels and decreased iNOS levels, compared to the control group. In the donkey milk kefir group, iNOS levels were significantly lower than those of the control and donkey milk groups, while eNOS levels were similar to the control group. Donkey milk kefir induced apoptosis, suppressed proliferation and decreased co-expression of iNOS and eNOS. Donkey milk promoted development of the tumors. Therefore, donkey milk kefir appears to be more beneficial for treating breast cancer than donkey milk.

Background: The inhibition of autophagy using pharmacological inhibitors such as chloroquine may be an effective strategy to overcome chemotherapy or resistance to anti-angiogenic therapy. Materials and Methods: The cytotoxic effect of doxorubicin (0.1-1 μM), chloroquine (0.25-32 μM) and their combination were investigated by employing ATP assay in human umbilical vein endothelial cells (HUVECs). The effect of doxorubicin and chloroquine combination was also measured using tube formation assay on Matrigel. The anti-angiogenic activities of doxorubicin (2.5 μg/pellet) and chloroquine (15 μg/pellet), their combination, and standards (50 μg/pellet) were tested in vivo using the chick embryo chorioallantoic membrane (CAM) assay. Results: The combination of doxorubicin and chloroquine significantly had a stronger anti-angiogenic effect than the positive control (±)-thalidomide and doxorubicin alone in the CAM assay and in vitro tube-formation assay. Conclusion: Chloroquine enhanced the anti-angiogenic effect of doxorubicin on CAM at the tested concentrations.

Rhus coriaria (sumac) naturally grows worldwide and contains many phytochemicals. Tannic acid is frequently used for treating medical conditions. The aim of this study is to determine the synergistic effects of methanolic and acetonic extracts of Rhus coriaria’s from unriped fruits in combination with tannic acid on angiogenesis by using the Chick Chorioallantoic Membrane (CAM) assay. The effects of methanol and acetone extracts of Sumac (MES and AES, respectively, at a final concentration of 5 mg/mL for both) and tannic acid (Tan 1.25 and 5 mg/mL) alone or in combination were examined for their effect on angiogenesis by applying the CAM assay. CAM assay results showed that AES is more effective than MES on suppressing angiogenesis. Tannic acid started to show antiangiogenic effects at the concentration of 5 mg/mL. When AES and tannic acid were used in combination at a dose of 5 mg/mL each, a very strong antiangiogenic effect was observed (score; 1.1±0.131, p<0.001). The combinatorial usage of AES and tannic acid may result in strong suppression of angiogenesis, an effect that is shown in this study for the first time. Based on this observation, combinatorial usage of sumac extracts and tannic acid could be promising as a new antiangiogenic therapy. Cite this article as: Üvez, A., Kılıç, Ş., Esener, O.B.B., Ünder, D., Saltan, F.Z., Dimas, K., Armutak, E.İ., 2021. Combinatorial Usage of Sumac Unriped Fruit Extract (Rhus Coriaria) and Tannic Acid Enhanced Synergistic Anti-Angiogenic Effect on Chick Chorioallantoic Membrane Assay. Acta Vet Eurasia 47, 2-9.