新疆塔里木马鹿茸多肽对小鼠前成骨细胞的影响

摘要

Objective To evaluate the effect of antler polypeptides from Cervus elaphus yarkandensis on MC3T3-E1 cells. Methods The concentration of antler polypeptides of Cervus elaphus yarkandensis was measured by BCA protein assay kit. MC3T3-E1 cells were cultured and analyzed by BCIP/NBT chromogenic alkaline phosphatase (ALP) staining kit. After being induced to form mineralized knot, the cells were stained by using Alizarin red staining. Three different concentrations (10, 1.0, 0.1 μg/mL) of antler polypeptides were analyzed by MTT method and micronutrients enzymes standard method to determine the effect of cell proliferation and ALP synthesis. Results The concentration of antler polypeptides was 0.07 mg/mL. The results of in vitro cell activity analysis showed that the positive rate of ALP was 90%and the mineralization knot was stained red. Compared with the control group, the different concentrations of antler polypeptides all showed the function of cell proliferation and the effect was dose-dependent after 3 d and 7 d. Compared with the control group, at the 3 d, three groups of antler polypeptides promoted synthesis and secretion of ALP (P<0.01) and the results showed a dose-dependent effect. Conclusion Antler polypeptides could obviously promote the proliferation of MC3T3-E1 cells and secretion of ALP, which indicated that antler polypeptides have certain effect on osteoporosis.%目的：探讨新疆塔里木马鹿茸多肽对小鼠前成骨细胞(MC3T3-E1)的作用机制。方法以BCA蛋白质定量试剂盒测定马鹿茸多肽样品的浓度；体外培养MC3T3-E1细胞，采用BCIP/NBT碱性磷酸酶(ALP)显色试剂盒染色；诱导分化形成矿化结节，进行茜素红染色；将MC3T3-E1细胞置于马鹿茸多肽高、中、低(10、1.0、0.1µg/mL)3个浓度的培养基中培养，采用MTT法和微量酶标法检测马鹿茸多肽不同浓度对MC3T3-E1细胞增殖和ALP合成的影响。结果 BCA 蛋白质定量检测结果显示，马鹿茸多肽蛋白浓度为0.07 mg/mL。体外培养 MC3T3-E1细胞ALP染色呈淡蓝色，阳性率为90%；矿化结染色呈红色。与对照组比较，第3、7日马鹿茸多肽高、中、低剂量组可促进MC3T3-E1细胞增殖，且呈现剂量依赖效应；第3日马鹿茸多肽高、中、低剂量组均可促进MC3T3-E1细胞分泌ALP(P＜0.01)，且呈现一定的剂量效应。结论塔里木马鹿茸多肽通过促进MC3T3-E1细胞增殖和ALP合成的分泌，达到治疗骨质疏松症的作用。