石榴籽油对雌激素相关骨质疏松症小鼠软骨代谢的保护作用及对相关蛋白表达影响的探究

Correlation metabolism study of affect of pomegranate seed oil on the osteoporosis cartilage mice

目的观察石榴籽油抑制骨质疏松症小鼠软骨代谢的效果。方法将36只雌性ddY小鼠随机分为6组:正常组、模型组、石榴籽油低、中和高剂量组以及雌二醇组,分别给予各组小鼠1g/kg/d饲料灌胃,于1、2、3、4周后检测各组小鼠的体质量变化以及4周后的子宫质量,观察正常组、模型组、不同PSO剂量实验组的骨质疏松病理切片情况,通过RT-PCR比较开始给予饲料以及1、2、3、4周后检测各组破骨细胞相关基因MMP2、CTR、Itgβ3以及成骨细胞相关基因Lrp5、OPN的表达水平。结果卵巢切除后模型组小鼠子宫质量显著下降;石榴籽油的低剂量对子宫质量的下降有显著抑制作用,其效果与雌二醇相当;石榴籽油的中剂量和大剂量对子宫质量的下降无明显抑制作用。模型组动物股骨近端骨小梁微结构破坏,程度最为严重,骨密质比较薄,骨细胞数量减少,骨小梁稀疏、断裂、排列紊乱,髓腔增大,测定其骨密度为(0.98±0.15)g/cm^3显著低于正常组(1.37±0.23)g/cm^3及PSO低剂量组(1.21±0.19)g/cm^3;同时PSO低剂量组骨质疏松的病理改变改善明显,骨小梁密增多,骨密质增厚。低剂量组(5%)石榴籽油对破骨细胞的三种基因(CTR、ITGb3、MMP2)呈现下调,其中CTR下调最为显著;而对成骨细胞相关基因Lrp5呈现显著上调,P<0.05。结论石榴籽油通过相关基因调节抑制破骨细胞活性,减轻骨吸收和破坏,从而抑制骨质疏松症小鼠骨量的丢失;同时通过作用于成骨细胞相关基因,促进成骨细胞分化,促进成骨。

Objective To study the molecular mechanism for Pomegranate seed oil in prevention of bone loss in a osteoporosis mice model.Methods 36 KM female mice randomly divided into 6groups which were normal group,model group,low,middle and high dose group of pomegranate seed oil.Mice were given 1g/kg/d feed to fill the stomach,after 1,2,3,and 4weeks,the changes of the body mass and the quality of the uterus were detected.And estradiol group,after 4 weeks detected the bone density,osteoporosis pathological sections were observed in normal group,model group and experimental groups.TRAP,CTR,ITGβ3,MMP2,OPN levels in the 1,2,3and 4 weeks were compared by RT-PCR.Results After the ovariectomy,the quality of the uterus in model group decreased significantly.Low dose of pomegranate seed oil had no significant effect on the decrease of uterine mass,the middle dose and high dose of pomegranate seed oil exhibit significant inhibition effect on the decrease of uterine mass and the effect was equivalent to the estradiol group.In the model group,the trabecular microstructure of the proximal femur was destroyed,the extent of which was the most serious,the bone density was relatively thin,the number of bone cells was reduced,bone fracture shows serious disorder with medullary cavity increased.The bone mineral density in model group was 0.98±0.15g/cm3 was significantly lower than that in the normal group 1.37±0.23g/cm3 and1.21±0.19g/cm3 in PSO low dose group.At the same time,the pathological changes of PSO low dose group were improved obviously,the density of trabecular bone was increased,and the bone density was increased.Control group animals damage degree of proximal femoral bone trabecular microstructure was most serious,followed by the experimental group,bone trabecular microstructure of blank group was normal.Low dose group（5%）,pomegranate seed oil of osteoclast three genes（CTR,ITGb3,MMP2）was presented the cut,cut the most significant one CTR.For osteoblasts related genes had a gene（Lrp5）was presented rise significantly,（P0.05）.Conclusion Pomegranate seed oil by related genes regulating inhibiting osteoclast activity,reduce bone resorption and damage,thus inhibiting osteoporosis mice bone mass loss.Through the effect on osteoblast genes at the same time,promote the osteoblast differentiation,promote osteogenesis.