巴戟天及其糖提取物对于体外培养成骨细胞OPG/RANKL基因系统表达的影响

目的:探讨巴戟天及多糖提取物对成骨细胞骨保护素(OPG)/核因子κB受体活化因子配体(RANKL)基因系统表达的影响。方法:取2~3天的SD大鼠5只分离原代成骨细胞,再取8周龄SD大鼠35只随机分为七组,对照组不进行处理,三组给予10 g/L、50 g/L、100 g/L巴戟天水灌胃,其余三组分别给予10 g/L、50 g/L、100 g/L巴戟天多糖灌胃,72 h后采用采用ELISA法测定培养液中OPG、RANKL及骨钙素的含量,采用MTT法检测不同浓度巴戟天水及多糖提取物对大鼠成骨细胞增殖的影响,采用荧光定量PCR检测OPG和RANKL mRNA表达情况;通过Westernblot检测OPG和RANKL蛋白表达水平。结果:巴戟天水及多糖提取物组A570nm、ALP活性、骨钙素含量、OPG/RANKL mRNA表达量、OPG和RANKL蛋白表达阳性密度均高于对照组(P0.05);A 570 nm、ALP活性、骨钙素含量、OPG/RANKL mRNA表达量、OPG和RANKL蛋白表达阳性密度均高于同等剂量的水提取物各组(P0.05);巴戟天多糖组中随着多糖剂量的升高A 570 nm、ALP活性、骨钙素含量、OPG/RANKL mRNA表达量、OPG和RANKL蛋白表达阳性密度,差异比较有统计学意义(P0.05)。结论:巴戟天水及多糖提取物均能促进体外培养成骨细胞的增殖,提高成骨细胞活性。

Effect of Morinda Root and its Polysaccharide Extract on OPG / RANKL Gene Expression of in vitro Osteoblast

Objective:To investigate effect of Morinda root polysaccharide and its extract on OPG/RANKL gene expression of invitro osteoblast, so as to make further research on molecular mechanisms of Morinda in the treatment of osteoporosis.Methods:Take 5SD rats birth from 2 to 3 days to separate primary osteoblasts, and then take 35 SD rats 8 weeks old randomly divided into 7 groups. Thecontrol group did not take treatment. Three polysaccharide groups were given separately 10 g / L, 50 g / L, 100 g / L Morinda watergavage, and the remain three groups were given separately 10 g / L, 50 g / L, 100 g / L Morinda polysaccharide gavage. After 72 h, theELISA assay was used to detect the contents of broth OPG, RANKL and bone calcium. MTT was applied to investigate the effect ofdifferent concentrations of Morinda officinalis water and polysaccharide extract on the proliferation of rat bone. And Real-time PCR wasused to detect the mRNA expression of OPG and RANKL. Western blot was applied to analyze the protein levels of OPG and RANKL.Results:In the Morinda officinalis water and polysaccharide extract groups, the A570 nm, ALP activity, osteocalcin content, OPG /RANKL mRNA expression, OPG and RANKL protein expression density were higher than in control group (P <0.05). The A570 nm,ALP activity, osteocalcin content, OPG / RANKL mRNA expression, OPG and RANKL protein expression density were also higher thanin each group of the same dose of water extract (P<0.05). In the Morinda officinalis polysaccharide groups, along with the increasingdoses of polysaccharide, A570 nm, ALP activity, osteocalcin content, OPG / RANKL mRNA expression, OPG and RANKL proteinexpression density all had statistically significant difference (P<0.05).Conclusion:The Morinda officinalis and its polysaccharide extractcan promote the in vitro proliferation of osteoblast cell, and enhance osteoblast activity.