Fig 2: Inhibition of autophagy with 3-MA reverses the protective effect of naringin. (a) The protein levels of LC3, Beclin-1, and P62 were measured by Western blotting. (b) The variation of autophagic flux was detected by confocal microscope (yellow spots represent autophagosomes, red spots represent autolysosomes). (c) TUNEL staining results to detect the rate of apoptosis in human NP cells. () The protein expressions of C-caspase-3, Bax, and Beclin-2 were measured by Western blotting. (e, g) The distribution of Collagen II (green), MMP-3 (red), and DAPI (blue) were detected by the immunofluorescence (scale bar: 100 μm). (f, h) The mRNA expressions of Collagen II were measure by qRT-PCR. (i) The protein expressions of Collagen II and MMP-3 were measured by Western blotting. The data represent the means ± SD. Significant differences between groups are indicated as ∗p < 0.05, ∗∗p < 0.01, and ∗∗∗p < 0.001, n = 3.

Fig 3: Expression of MMP-3 in the periodontal tissue in the diabetes group. MMP-3 was positively expressed in (A) the gingival epithelial layer, (B) the granular layer and stratum spinosum, (C) fibroblasts, (D) alveolar bone cells, (E) the cytoplasm of bone marrow stromal cells and (F) the gingival epithelial basal layer. Magnification, ×400. MMP, matrix metalloproteinase.

Fig 4: Expression levels of TNF-α and MMP-3 in the periodontal tissue in the diabetes + β-anhydroicaritin group. TNF-α was marginally positively expressed in (A) the gingival epithelial layer and (B) fibroblasts. MMP-3 was marginally positively expressed in (C) the gingival epithelial layer and (D) fibroblasts. Magnification, ×400. TNF, tumor necrosis factor; MMP, matrix metalloproteinase.

Fig 5: a Mild expression of MMP-3 is present in the cytoplasm of tumor cells, especially those with a higher degree of anaplasia. (×200) b Expression of p-IGF1R was seen in tumoral cells. (×400) c Expression of p-IGF1R was seen in tumoral cells. (×200)