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Objective:To observe the effect of Ginsenoside Re on the proliferation and protein secretion of primary cardiac fibroblasts (CFs) cultured in high glucose by vitro, and the regulation of Wnt/β-catenin signaling pathway.Methods:The myocardial fibroblast proliferation model induced by high glucose in vitro was used. Cell proliferation was detected by MTT method, cell cycle was measured by flow cytometry, concentration of type Ⅰ,Ⅲ collagens and TGF-β 1 protein were tested by ELISA assay. Protein expression of β-catenin, GSK-3β and p-GSK-3β were determined by Western blot. Results:Compared with the model group, the cell proliferation in Ginsenoside Re high, medium, low group were significantly decreased ( P<0.01), the percentage of cells in G 0 + G 1 phase was increased ( P<0.01), and the percentage of cells in S + G 2 + M phase was decreased ( P<0.01), the content of TGF-β 1 was significantly decreased( P<0.01). The content of type Ⅲ collagen [(6.566±1.620)ng/ml,(7.170±0.470)ng/ml vs. (11.241±2.234)ng/ml] in Ginsenoside Re high, medium group were significantly decreased ( P<0.01). The expression of β-catenin (0.281±0.016, 0.301±0.021 vs. 0.409±0.037) was significantly decreased and the expression of p-GSK-3β (0.369±0.049 vs. 0.268±0.048) in Ginsenoside Re high, medium group were significantly increased ( P<0.01). Conclusion:Ginsenoside Re plays an important role in inhibiting CFs proliferation and reducting the synthesis of collagen and TGF-β 1 by regulating abnormal expression of Wnt/β-catenin signaling pathway. It has the potential to delay the myocardial fibrosis of diabetes mellitus.
RESUMO
Objective To observe the effects of dehydrocorydaline (DHC) on proliferation and collagen secretion of cardiac fibroblasts (CFs) cultured by high glucose;To provide experimental evidence for clinical application of Rhizoma Corydalis. Methods CFs cultured in vitro with high glucose were made into models. Collagenase and trypsin were used for the combine digestion of CFs from ratsbornin 24 h. 2-4 generation CFs were cultured by high glucose (25 mmol/L), and then 100, 50, 25 mg/L dethydrocorydaline was added for intervention. Cellular morphology of CFs was observed after 24, 48 h. CFs proliferation was detected by MTT method. Cell cycle was assessed via flow cytometry. The levels of collagen Ⅰ and collagen Ⅲ were determined by ELISA. Results CFs began to grow adherence 3 hours after planting, and CFs cultured by high glucose significantly proliferated 24, 48 h later (P<0.05, P<0.01). The percentage of S+G2+M phase CFs increased significantly after 48 h (P<0.01). The secretion of collagen Ⅰ and collagen Ⅲ also increased significantly (P<0.01). After the intervention of DHC, CFs proliferation was significantly inhibited (P<0.01);the percentage of S+G2+M phase CFs decreased (P<0.01);the secretion of collagen Ⅰ and collagen Ⅲ was reduced (P<0.05, P<0.01). Conclusion DHC can reduce CFs proliferation, decrease collagen secretion of CFs cultured by high glucose, and has potential effects of anti-myocardial fibrosis.