Effect and mechanism by which Pterocarya hupehensis skan total flavonoids regulates the proliferation, migration and apoptosis of fibroblast-like synoviocytes

doi:10.12307/2026.547

Abstract

Abstract: BACKGROUND: Studies have confirmed that Pterocarya hupehensis skan total flavonoids (PHSTF) can improve the level of collagen-induced arthritis in rats, but there is still a lack of research on the regulation of Wnt/β-catenin signaling pathway in fibroblast-like synoviocytes and its effect on related cell functions.
OBJECTIVE: To investigate the effect and mechanism of PHSTF on lipopolysaccharide-induced proliferation, migration and apoptosis of fibroblast-like synoviocytes based on the Wnt/β-catenin signaling pathway
METHODS: Fibroblast-like synoviocytes were divided into control group, lipopolysaccharide group, lipopolysaccharide + low-, medium-, and high-dose PHSTF groups (10, 20, and 40 μg/mL), lipopolysaccharide + Wnt pathway inhibitor DKK1 group, and lipopolysaccharide + Wnt pathway inhibitor DKK1 + high-dose PHSTF group (40 μg/mL). The cell counting kit-8 method was used to detect the effect of PHSTF on the viability of fibroblast-like synoviocytes, and the final drug concentration and time were screened. Flow cytometry was used to detect the apoptosis of fibroblast-like synoviocytes. Cell scratch assay, EDU staining and cell cloning assay were used to detect the migration and proliferation of fibroblast-like synoviocytes. Western blot assay was used to detect the protein expression levels of Wnt3a, β-catenin, tumorigenic genes, matrix metalloproteinase 2, matrix metalloproteinase 9, Bax and Bcl-2 in fibroblast-like synoviocytes.
RESULTS AND CONCLUSION: (1) Compared with the control group, the cell viability decreased significantly when the concentration of PHSTF was > 40 μg/mL
(P < 0.01). Therefore, the drug concentration of ≤ 40 μg/mL was selected for subsequent experiments. (2) Compared with the lipopolysaccharide group, the wound healing rate, cell clone formation rate and the number of EDU-positive cells in the low-, medium- and high-dose PHSTF groups were significantly reduced, while the apoptosis rate was significantly increased (P < 0.05-0.01). (3) Western blot results showed that compared with the lipopolysaccharide group, low-, medium- and high-dose PHSTF significantly inhibited cellular Wnt3a, β-catenin, cellular tumorigenic genes, matrix metalloproteinase 2, matrix metalloproteinase 9, and Bcl-2 protein expression, and promoted the expression of Bax protein (P < 0.01). (4) Compared with the DKK1 group, the combination of DKK1 and high-dose PHSTF significantly inhibited the protein expression of Wnt3a, β-catenin, matrix metalloproteinase 2, matrix metalloproteinase 9 and Bcl-2 protein expression and promoted the protein expression of Bax (P < 0.01). To conclude, PHSTF may inhibit the proliferation and migration of fibroblast-like synoviocytes and promote apoptosis by inhibiting the Wnt/β-catenin signaling pathway.

Key words: Pterocarya hupehensis skan total flavonoids, fibroblast-like synoviocytes, rheumatoid arthritis, Wnt/β-catenin, apoptosis, proliferation

CLC Number:

Bao Zhuoma, Hou Ziming, Jiang Lu, Li Weiyi, Zhang Zongxing, Liu Daozhong, Yuan Lin. Effect and mechanism by which Pterocarya hupehensis skan total flavonoids regulates the proliferation, migration and apoptosis of fibroblast-like synoviocytes[J]. Chinese Journal of Tissue Engineering Research, 2026, 30(4): 816-823.

Figures/Tables 8

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