Type 2 diabetes mellitus is on the increase worldwide. Current treatments possess undesirable side-effects and therefore investigations into alternative remedies, which may be cost-effective and devoid of such side-effects, are ongoing. Aqueous and methanol extracts of the bark of Sclerocarya birrea (A. Rich.) Hochst. subsp. caffra (Sond.) Kokwaro (Anacardiaceae) and Ziziphus mucronata Willd. subsp. mucronata (Rhamnaceae) were subjected to testing in order to assess their in vitro anti-diabetic activity. Total polyphenolic content and antioxidant activity, as well as the effect on α-amylase, α-glucosidase and cytotoxicity, were measured. Furthermore, glucose uptake was evaluated in C2C12 myotubes, 3T3-L1 adipocytes and HepG2 hepatocarcinoma cells, and insulin secretion in RIN-m5F rat pancreatic β-cells. The extracts of both plants inhibited the activities of α-amylase and α-glucosidase in a concentration dependent manner, with results being comparable with the positive control, acarbose. All the extracts contained antioxidant activity, with the methanol extracts of S. birrea displaying the strongest free radical scavenging capacity (IC 50 = 2.16 µg/mL). Most of the extracts were not cytotoxic to the cell lines with IC 50 values > 100 µg/mL. Extracts of both plants significantly increased glucose uptake in C2C12, 3T3-L1 and HepG2 cells. However, insulin secretion from RIN-m5F cells was not altered. This study provides evidence that the plants possess in vitro anti-diabetic properties.