摘要
【目的】观察中药骨康含药血清对肾阳虚型骨质疏松症患者成骨细胞细胞因子骨保护素(OPG)、核因子-κB受体活化因子配体(RANKL)mRNA表达的影响。【方法】取6月龄体质量2.5～3.0 kg的新西兰大耳白兔4只,雌雄各半,随机分为2组(中药骨康组、空白对照组),每组2只;分别灌服骨康煎剂4.2 g.kg-1.d-1,以及等容积生理盐水,连续灌胃共7 d,最后1次灌胃2 h后所有动物进行心脏采血,收集血清备用。分离并传代培养肾阳虚骨质疏松症患者成骨细胞,取第2代,制成2×105/mL的细胞悬液。实验组分别加入低、中、高不同浓度(体积分数5%、10%、20%)的骨康含药血清进行干预,空白对照组仅加培养液,继续培养。48 h后采用Trizol一步法提取总RNA,然后采用逆转录—聚合酶链反应(RT-PCR)法检测OPG、RANKL mRNA的表达。【结果】骨康低、中、高浓度组OPG mRNA表达量显著上升,与空白对照组比较差异均有统计学意义(P<0.05或P<0.01);骨康中浓度组与骨康低浓度组、骨康高浓度组与骨康中浓度组组间分别比较差异均有统计学意义(P<0.05);骨康含药血清以体积分数5%、10%、20%浓度递增时,RANKL mRNA的表达量显著下降,与空白对照组比较差异均有统计学意义(P<0.05),骨康中浓度组与骨康低浓度组、骨康高浓度组与骨康低浓度组组间RANKL mRNA表达量比较差异均无统计学意义(P>0.05)。【结论】骨康可以上调肾阳虚骨质疏松症患者体外培养成骨细胞OPG mRNA的表达,下调RANKL mRNA的表达。
Objective To observe the effect of serum （OPG） and receptor activator of nuclear factor kappa-B containing Chinese medicine Gukang on oxypolygelatin ligand （RANKL） mRNA expression in in-vitro cultured human osteoblasts from osteoporosis patients with kidney-yang deficiency. Methods Four six-month-old new Zealand rabbits with the weight of 2.5 - 3.0 kg, male in half, were equally randomized into two groups, Gukang group （4.2 g.^-1·d^-1） and blank control group （the same volume of normal saline ）. The medication by gastric garage lasted 7 days. Blood sampling via cardiac puncture was carried out 2h after the last gastric gavage for the preparation of serum. Osteoblasts were isolated from osteoporosis patients with kidney-yang deficiency and then were subcuhured. The second generation of subcultured osteoblasts was made into cell suspension at the density of 2 ~ 105/mL. Serum containing Chinese medicine Gukang at the concentrations of 5%, 10% and 20% was added into the experiment group for culture, and the ceils in the blank control group were cultured with the basic culture solution. After culture for 48 h, total RNA was extracted from the cultured osteoblasts cells by the method of one- step Trizol, and then the mRNA expression of OPG and RANKL was examined by reverse transcriptase polymerase chain reaction （RT-PCR）. Results OPG mRNA expression level was higher in low-, moderate and high-dosage Gukang groups than that in the blank controlgroup （P 〈0.05 or P 〈0.01 ）, and the differences between the three groups were significant （ P 〈 0. 05 ）. RANKL mRNA expression level was decreased in Gukang groups with the increase of Gukang-containing serum concentration （P 〈 0. 05 compared with that in the blank control group）, but the differences between the three groups were insignificant （ P 〉 0. 05 ）. Conclusion Serum containing Gukang exerts certain effect on up-regulating OPG mRNA expression and on down-regulating RANKL mRNA expression in in-vitro cultured human osteoblasts
出处
《广州中医药大学学报》
CAS
北大核心
2012年第4期392-396,487,共6页
Journal of Guangzhou University of Traditional Chinese Medicine
基金
国家自然科学基金资助课题(编号:30600830)
