The present study aimed to investigate the effect of β-anhydroicaritin on the expression levels of tumor necrosis factor (TNF)-α and matrix metalloproteinase (MMP)-3, and the pathological changes in the periodontal tissue of diabetic rats. Male Wistar rats (n=40; three months old) were randomly divided into four groups: Normal control group, diabetes group, diabetes + β-anhydroicaritin group and diabetes + urate group, (n=10 in each group). Following an overnight fast, diabetes was induced by intraperitoneal injection of streptozocin. The rats were maintained for 12 weeks and the blood sugar, urine sugar and body weight were assessed in week 12. Histological changes of the periodontal tissues were observed by hematoxylin and eosin staining, and the expression levels of TNF-α and MMP-3 were observed by immunohistochemistry. Following 12 weeks, the TNF-α grey value in the diabetes group was significantly lower compared with that in the control group (P<0.05), while no significant difference was observed between TNF-α levels in the diabetes + β-anhydroicaritin group, diabetes + urate group and the control group (P>0.05). However, TNF-α levels in the diabetes + β-anhdroicaritin group and diabetes + urate group were significantly higher compared with those in the diabetes group (P<0.05), and those in the diabetes + β-anhydroicaritin group were lower compared with those in the diabetes + urate group (P<0.05). The MMP-3 grey value in the diabetes group was significantly lower compared with that in the control group (P<0.05), while no significant difference was observed between MMP-3 levels in the diabetes + β-anhydroicaritin group, diabetes + urate group and the control group (P>0.05). However, MMP-3 levels the diabetes + β-anhydroicaritin group and diabetes + urate group were significantly higher compared with those in the diabetes group (P<0.05), and those in the diabetes + β-anhydroicaritin group were lower compared with those in the diabetes + urate group (P<0.01). β-anhydroicaritin normalized the expression levels of TNF-α and MMP-3 in the periodontal tissue of diabetic rats and led to the recovery of the changes in the morphological structure of the periodontal tissue.