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目的:探讨葛根素(Pue)对高糖培养的成骨细胞增殖、分化及Ⅰ型胶原m RNA(CollⅠm RNA)表达的影响,为临床治疗糖尿病骨质疏松症提供实验依据。方法:取出生24 h内的SD大鼠的颅骨,体外分离培养成骨细胞,待细胞形态稳定后,取对数生长期的成骨细胞,随机分为正常组、高糖组(葡萄糖浓度22 mmol/L)、葛根素各浓度组(葛根素浓度分别为10-8 mol/L、10-7mol/L、10-6mol/L)、高糖加葛根素各浓度组,每组设8个复孔,分别培养48 h,隔天换液并加药。测定成骨细胞的增殖能力、细胞内ALP活性以及CollⅠm RNA表达情况。结果:与正常组比较,葛根素各浓度组细胞增殖、细胞ALP活性、细胞CollⅠm RNA表达均显著增高,差异均有统计学意义(P<0.05,P<0.01);高糖组、高糖+葛根素各浓度组明显低于正常组,差异均有统计学意义(P<0.05,P<0.01)。与高糖组比较,高糖+葛根素各浓度组细胞增殖、细胞ALP活性、细胞CollⅠm RNA表达均显著增高,差异均有统计学意义(P<0.05,P<0.01)。结论:高糖情况下抑制成骨细胞的增殖、分化和Ⅰ型胶原表达,但加入葛根素后能提高成骨细胞的增殖、分化和Ⅰ型胶原表达,说明葛根素对糖尿病骨质疏松症有治疗作用。
OBJECTIVE: To investigate the effects of Puerarin on the proliferation and differentiation of cultured osteoblasts and the expression of Coll Ⅰm RNA in osteoblasts cultured with high glucose, and to provide experimental evidence for clinical treatment of diabetic osteoporosis. Methods: The skulls of SD rats within 24 h after birth were removed and osteoblasts were isolated and cultured in vitro. After the cells were stable, the logarithmic growth phase osteoblasts were randomly divided into normal group, high glucose group (glucose concentration 22 (p <0.05), puerarin concentration (10-8 mol / L, 10-7 mol / L, 10-6 mol / L) Replenishment, were cultured for 48 h, the next day for fluid and dosing. The proliferation of osteoblasts, intracellular ALP activity and Coll Ⅰm RNA expression were measured. Results: Compared with the normal group, the cell proliferation, cell ALP activity and Coll Ⅰm RNA expression were significantly increased in all concentrations of puerarin (P <0.05, P <0.01) Puerarin concentrations were significantly lower than the normal group, the difference was statistically significant (P <0.05, P <0.01). Compared with high glucose group, cell proliferation, cell ALP activity and Coll Ⅰm RNA expression were significantly increased in high glucose + puerarin groups, with statistical significance (P <0.05, P <0.01). CONCLUSION: The proliferation, differentiation and type Ⅰ collagen expression of osteoblasts were inhibited under high glucose condition. However, the addition of puerarin increased the proliferation and differentiation of osteoblasts and the expression of type Ⅰ collagen, indicating that puerarin has effects on diabetic osteoporosis Therapeutic effect.