三七总皂甙对大鼠骨髓基质细胞OPG/RANKL表达比的影响

目的观察三七总皂甙（PNS）对大鼠骨髓基质细胞中骨保护素（OPG）/核因子Kappa B受体活化因子配体（RANKL）表达的影响,探讨三七总皂甙防治骨质疏松的作用机制。方法分离、培养大鼠骨髓基质细胞（BMSCs）,取P3代细胞,分成4组,各PNS组终浓度分别为50、100、200 mg/L;对照组不进行特殊处理。培养7 d后,采用RT-PCR和Western Blotting检测OPG、RANKLmRNA和蛋白的表达并计算OPG/RANKL比值。结果 RT-PCR显示：PNS组（50、100、200 mg/L）和对照组OPG/RANKLmRNA表达比分别为（0.40±0.07）、（0.41±0.08）、（0.78±0.11）和（0.23±0.05）;蛋白印迹实验显示PNS组（50、100、200 mg/L）和对照组OPG/RANKL蛋白表达比分别为（0.25±0.06）、（0.64±0.13）、（0.28±0.09）和（0.13±0.03）。在mRNA和蛋白水平,PNS组OPG/RANKL比值均明显高于对照组,100 mg/L PNS组最高,差异有统计学意义（P〈0.05）。结论三七总皂甙可增加骨髓基质细胞中OPG的表达,减少RANKL的表达。

Effect of Total Saponins of Panax Notoginseng on Expression of Osteoprotegerin/receptor of Zctivator of Nuclear RactorK B-ligand in Rat Bone Marrow Stromal Cells

Objective To evaluate the effect of total Saponins of Panax Notoginseng （PNS） on the expression of osteoprotegerin （OPG） /receptor of activator of nuclear factorKB-ligand （RANKL） expression in rat bone marrow stromal cells （BMSCs） , and to investigate the mechanism of it on prevention and treatment for osteoporosis. Methods BMSCs were isolated from femur and tibia of adult rats. After 2 passages, BMSCs were divided into 4 groups. PNS groups were treated with different concentrations of PNS （50, 100, and 200 rag/L） while in control group, there was no special treatment. 7 d after treatment, the expression of OPG and RANKL were evaluated by RT-PCR and Western Blotting. Results In RT-PCR, the ratio of OPG/ RANKL in PNS groups and control group were （0.40 ~ 0.07 ）, （0.41 ＋ 0. 08 ）, （0.78 ＋ 0.11 ） and （0.23 ＋ 0.05 ）, respectively. In Western Blotting the ratio of OPG/RANKL in PNS groups and control group were （0.64 ＋ 0.13 ）, （0.28 ＋ 0.09）, （0.13 ~ 0.03 ）, and （0.25 ~ 0.06）, respectively. The ratio of OPG/RANKL were higher in PNS groups than it in control group, with 100 mg/L PNS group is the highest （P 〈 0.05 ）. Conclusion Total Saponins of Panax Notoginseng may enhance OPG expression and inhibit RANKL expression of rat bone marrow stromal cells.