骨碎补总黄酮调控miR-195-5p干预TGF-β1对大鼠生长板软骨细胞增殖的影响OA

Objective To investigate the molecular mechanism by which total flavonoids of rhizoma drynariae affect the proliferation of growth plate chondrocytes in rats,specifically focusing on their effect on TGF-β1 expression in the Ihh-PTHrP pathway via modulating miR-195-5p.Methods Chondrocytes were harvested from the proximal tibial growth plate of two-week-old SD rats,then cultured and identified.Subsequently,in vitro intervention was performed using different concentrations of total flavonoids of rhizoma drynariae(5,20,100,250,500 mg/L).The selection of the optimal drug concentration was carried out using the MTT method,and the screening of plasmids inhibiting PTHrP expression was done by qRT-PCR technology.MTT method and HE staining were employed to evaluate the effect of total flavonoids of rhizoma drynariae and PTHrP on cell proliferation.The expression level of TGF-β1 was determined by ELISA.The target genes of miR-195-5p were validated through bioinformatics analysis and dual-luciferase reporter gene assays.Results The results showed that a concentration of 250 mg/L of total flavonoids of rhizoma drynariae had the best effect on promoting cell proliferation.Both total flavonoids of rhizoma drynariae and PTHrP significantly enhanced cell proliferation,while PTHrP inhibitors had the opposite effect.The differential expression of miR-195-5p and its regulatory role on the expression of TGF-β1 were verified.Conclusion Total flavonoids of rhizoma drynariaemaypromote the proliferation of growth plate chondrocytes by downregulating the expression of miR-195-5p,thereby upregulating TGF-β1 protein expression.