论文部分内容阅读
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(P<0.05);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(P<0.05);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(P<0.05)。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。
Objective: To investigate the effect of Morinda officinalis and polysaccharide extracts on the expression of osteoblast osteoprotegerin (OPG) / nuclear factor κB receptor activator ligand (RANKL) gene system. Methods: Five primary osteoblasts were isolated from 5 SD rats of 2 ~ 3 days. 35 SD rats of 8 weeks old were randomly divided into seven groups, the control group was not treated, the three groups were given 10 g / L, 50 g / L, 100 g / L Morinda officinalis Tianshui gavage, and the remaining three groups were given 10 g / L, 50 g / L, 100 g / L Morinda polysaccharides gavage 72 h later by ELISA assay culture The contents of OPG, RANKL and osteocalcin in liquid were determined by MTT assay. The effects of different concentrations of Morinda officinalis and polysaccharide extract on the proliferation of osteoblasts were detected by MTT assay. The mRNA expression of OPG and RANKL was detected by real-time PCR. OPG and RANKL protein expression levels. Results: The A570nm, ALP activity, osteocalcin content, OPG / RANKL mRNA expression, OPG and RANKL positive expression of Morinda officinalis and polysaccharide extract group were higher than those of the control group (P <0.05); A 570 nm, ALP The activity, osteocalcin content, OPG / RANKL mRNA expression, OPG and RANKL protein expression positive density were higher than the same dose of water extract (P <0.05); Morinda officinalis polysaccharide group with the polysaccharide dose rise High A 570 nm, ALP activity, osteocalcin content, OPG / RANKL mRNA expression, OPG and RANKL protein expression positive density, the difference was statistically significant (P <0.05). Conclusion: Morinda officinalis and polysaccharides extract can promote the proliferation of cultured osteoblasts in vitro and increase the activity of osteoblasts.