枸杞多糖含药血清对MC3T3-E1细胞内Ca^(2+)及Ⅰ型胶原蛋白合成的影响

Effects of serum containing Lycium barbarum polysaccharides on the intracellular Ca^(2+) concentration and the expression of collagen type Ⅰ in MC3T3-E1 cells in vitro

背景:前期实验发现枸杞多糖对成年去势雌性大鼠骨质疏松有明显的骨质改善作用。目的:观察含枸杞多糖大鼠血清对成骨细胞株MC3T3-E1细胞内Ca2+及Ⅰ型胶原蛋白合成的影响。方法:将20%空白血清(对照组),5%,10%,20%含枸杞多糖血清分别加入成骨细胞株MC3T3-E1细胞培养液中,通过MTT法测定细胞增殖,激光扫描共聚焦显微镜测定胞内游离钙离子浓度,免疫细胞化学方法检测Ⅰ型胶原蛋白表达。结果与结论:与对照组比较,5%,10%含枸杞多糖血清可明显促进成骨细胞株MC3T3-E1细胞增殖(P<0.05,P<0.01);10%,20%含枸杞多糖血清组成骨样细胞株MC3T3-E1内细胞内Ca2+水平及Ⅰ型胶原蛋白合成明显升高(P<0.05,P<0.01)。说明含枸杞多糖血清可能通过影响细胞内Ca2+水平及Ⅰ型胶原蛋白合成防治骨质疏松症。

BACKGROUND： Previous experiments have shown that Lycium barbarum polysaccharide has a remarkable therapeutic effect on osteoporosis in adult castrated female rats. OBJECTIVE： To explore the effects of rat serum containing Lycium barbarum polysaccharides on the intracellular Ca2＋ concentration and the expression of collagen type Ⅰ in osteoblastic MC3T3-E1 cells in vitro. METHODS： The rat serum containing 5%, 10% and 20% Lycium barbarum polysaccharide was added into the media of osteoblastic MC3T3-E1 cells, respectively. Blank serum with 20% volume concentration was added into the control group. 3-（4,5-Dimethylthiazol-2-yl）-2,5-diphenyltetrazolium bromide viability analysis was utilized to investigate the cell proliferation. The intracellular Ca2＋ concentration was determined by laser scanning confocal microscope. The expression of collagen type Ⅰ was examined by immunocytochemical technique. RESULTS AND CONCLUSION： Compared with control group, the rat serum containing 5% and 10% Lycium barbarum polysaccharides significantly promoted the proliferation of osteoblastic MC3T3-E1 cells （P 〈 0.05, P 〈 0.01）. Rat serum containing 10% and 20% Lycium barbarum polysaccharides increased the intracellular Ca2＋ concentration and the expression of collagen typeⅠin osteoblastic MC3T3-E1 cells （P 〈 0.05, P 〈 0.01）. These findings indicate that rat serum containing Lycium barbarum polysaccharides may prevent and heal osteoporosis by affecting the intracellular Ca2＋ concentration and the expression of collagen typeⅠ.