中文摘要：

目的：观察中药单体梓醇在成骨-破骨共育体系中对成骨细胞（OB）增殖、OB碱性磷酸酶（ALP）活性、破骨细胞（OC）活性及OB雌激素受体（ER）α及βmRNA表达的影响,从细胞水平阐释其防治骨质疏松症的作用机制。方法：分别选取1 d和5 d SD大鼠分离培养OB和OC,并建立OB-OC共育体系;在共育体系中,用MTT法检测低浓度（0.05、0.1、0.5、1 mg/L）、中浓度（2、5、10 mg/L）和高浓度（20、50和100 mg/L）梓醇干预下的OB增殖率,并以梓醇最佳促OB增殖浓度进行后续实验,实验分为对照组和梓醇组。p NPP法检测各组OB的ALP活性;光镜观察OC骨吸收陷窝数目;重氮盐法检测OC抗酒石酸酸性磷酸酶（TRAP）的活性;RT-PCR方法检测OB ERα及ERβmRNA的表达。结果：在OB-OC共育体系中,0.05~2 mg/L梓醇各组中OB的增殖率显著高于对照组（P〈0.01）,且0.05 mg/L梓醇组促进OB增殖的能力明显高于其它浓度组（P〈0.01）,5~100 mg/L梓醇各组OB的增殖率与对照组比较无显著差异（P〉0.05）。0.05 mg/L梓醇组的ALP水平高于对照组（P〈0.05）,并在作用48、72和96 h后均对OC骨吸收陷窝数及TRAP有明显抑制作用（P〈0.01）;0.05 mg/L梓醇组OB中ERαmRNA的表达与对照组相比差异无统计学意义（P〉0.05）,而ERβmRNA的表达显著高于对照组（P〈0.05）。结论：梓醇在共育体系中可提高OB增殖和成骨活性,抑制OC活性并上调OB中ERβmRNA的表达。

英文摘要：

AIM： To investigate the effect of catalpol on the activity of osteoblasts（ OB） and osteoclasts（ OC）,and OB estrogen receptor（ ER） α/β mRNA expression in the OB-OC co-culture system. METHODS： OB and OC were isolated from the SD rats of 1 and 5 days old. In the OB-OC co-culture system,different concentrations of catalpol including low dosage（ 0. 05,0. 1,0. 5 and 1 mg / L）,middle dosage（ 2,5 and 10 mg / L）,and high dosage（ 20,50 and100 mg / L） were added into the culture medium to detect the changes of OB proliferation by MTT assay. The catalpol at maximal dosage was added to OB section to detect the alkaline phosphatase（ ALP） activity of OB by p NPP method. The mRNA expression of ERα / β in the OB treated with catalpol in the co-culture system was detected by RT-PCR. The catalpol at maximal dosage was added to OC group to detect the activity of OC by microscopy and tartrate-resistantacid phosphatase（ TRAP） activity detection. RESULTS： In 0. 05 ~ 2 mg/L catalpol groups,the proliferation of OB was significantly increased as compared with control group in the co-culture system,and it reached the maximum value when catalpol was at0. 05 mg / L,while in 5 ~ 100 mg / L catalpol groups,the proliferation of OB was not increased. The ALP activity of OB in0. 05 mg / L catalpol group was higher than that in control group. The catalpal at 0. 05 mg / L promoted the mRNA expression of ERβ in OB in the co-culture system,but did not increase the mRNA expression of ERα as compared with control group.Catalpol at 0. 05 mg / L obviously inhibited the bone resorption and the TRAP activity in OC. CONCLUSION： Catalpol stimulates the proliferation and activity of OB,inhibits the bone resorption and activity of OC,and increases the mRNA expression of ERβ in OB in the OB-OC co-culture system,suggesting that high mRNA expression of ERβ may be the regulatory pathway of catalpol in response to bone metabolism.