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目的研究中药骨康方对骨质疏松模型大鼠血清及骨组织中ERα和ERβm RNA表达水平的影响。方法选取4周龄健康雌性SD大鼠25只,随机分为阴性组、模型组、5%加药组、10%加药组、20%加药组,每组5只。除阴性组外,其余大鼠均进行双侧卵巢切除,手术后12周,5%、10%、20%加药组分别以5%、10%、20%剂量骨康方灌胃3周后处死,分别取血清及股骨标本。应用MTT法检测骨康方对成骨细胞株的增殖作用,采用RT-q PCR检测ERα和ERβm RNA的表达水平。结果与阴性组相比,中药骨康方浓度5%~20%范围内呈剂量依赖性促进成骨细胞株的增殖,且20%药物血清中培养24 h增殖最显著,因此将20%加药组作为测定大鼠ERα和ERβ表达变化组。阴性组血清与股骨干骺ERα和ERβm RNA均有表达;模型组ERα和ERβm RNA表达量较低;20%加药组ERα和ERβm RNA表达水平较模型组明显升高,ERβm RNA水平的增高尤为明显,更接近阴性组,差异均有统计学意义(P<0.05)。结论骨康方对骨质疏松患者的治疗作用可能是通过上调ERβ的表达水平,对骨组织进行保护,起到治疗骨质疏松作用,为临床应用提供药效学依据。
Objective To study the effects of Gukang Prescription on the expression of ERα and ERβmRNA in serum and bone tissue of osteoporosis model rats. Methods Twenty-five healthy female Sprague Dawley rats aged 4 weeks were randomly divided into 5 groups: negative group, model group, 5% plus drug group, 10% plus drug group and 20% plus drug group. Except for the negative group, all the other rats underwent bilateral ovariectomy. After 12 weeks of operation, the rats in 5%, 10% and 20% dosing groups were treated with 5%, 10% and 20% Sacrificed, were taken serum and femur specimens. The proliferation of osteoblast cells was detected by MTT assay. The expression of ERα and ERβmRNA was detected by RT-q PCR. Results Compared with the negative control group, the proliferation of osteoblast cells was promoted in a dose-dependent manner in the range of 5% ~ 20% at the concentration of Gukang decoction, and the proliferation was the most obvious at 20% Group as a measure of change in rat ERα and ERβ expression group. The expressions of ERα and ERβmRNA in the serum of the negative group and the epiphysis of the femoral shaft were all lower than those in the untreated group. The expressions of ERα and ERβmRNA in the model group were lower than those in the untreated group. The levels of ERα and ERβmRNA were significantly increased Obviously, closer to the negative group, the difference was statistically significant (P <0.05). Conclusion Gukang decoction can improve osteoporosis in patients with osteoporosis by up-regulating the expression of ERβ, protecting bone tissue, treating osteoporosis and providing pharmacodynamic evidence for clinical application.