“承载丸”对股骨头坏死大鼠细胞黏附分子基因的影响

Effect of Chengzai pill on the genes of cell adhesion molecules in rats with femoral head necrosis

背景:研究表明承载丸治疗激素性股骨头坏死具有明确的疗效,可明显增加骨密度、骨重量、骨强度和刚性,能改变雌激素水平低下状态。目的:观察承载丸对激素性股骨头坏死大鼠骨细胞中细胞黏附分子基因的影响,以明确该药对坏死股骨头内血管恢复正常血运的作用。设计、时间及地点:分组对照观察,于2006-03/08在中国中医科学院望京医院药理实验室及生物芯片北京国家工程研究中心所属博奥芯片公司实验室完成。材料:6月龄雄性SD大鼠6只,体质量(280±20)g。承载丸主要成分:有当归,杜仲,黄芪,枸杞子,鹿角霜,肉苁蓉、地鳖虫,水蛭、丹参,续断等22味中药。由北京市勃然制药有限公司提供。方法:6只SD大鼠采用内毒素和甲基强地松龙制做激素性股骨头坏死动物模型,随机将大鼠分为模型组和承载组,每组各3只。承载组大鼠自第1次注射甲基强地松龙后,灌服承载丸药液1.5g/kg,1次/d,共灌服6周。6周后取股骨头提取总RNA,进行基因谱测试。主要观察指标:基因谱测试结果中的细胞黏附分子基因作用路径分析。结果:3只模型大鼠使用承载丸后,与模型大鼠相比较,出现1.5倍以上改变的基因,分别为633个(下调506个,上调127个)、883个(下调640个,上调243个)、593个(下调408个,上调185个)。使用MAS软件归类分析后,共有相关作用路径79个,涉及297个基因。其中细胞黏附分子路径有8个下调基因。结论:大鼠服用承载丸后,使上述基因下调,恢复了大鼠巨噬细胞及靶细胞的识别功能,恢复了上皮细胞包括血管内皮细胞的正常生存条件,细胞黏附分子作用路径持正常水平,这是坏死股骨头内血管恢复正常血运的关键。

BACKGROUND： Studies have showed that Chengzai pill is of distinct significance in the treatment of steroid-induced osteonecrosis of femoral head through increasing bone mineral density, bone weight, bone strength and rigidity, it also reverses the low levels of estrogen. OBJECTIVE： To explore the effect of Chengzai pill on the genes of cell adhesion molecules on bone cells of steroid-induced osteonecrotic rats, and to understand the role to resume normal blood transport in femoral head of steroid-induced osteonecrosis. DESIGN, TIME AND SETTING： Grouping controlled observation was performed in the Pharmacological Laboratory of Wangjing Hospital of China Academy of Chinese Medical Sciences and laboratory of CapitalBio Corporation between March and August in 2006. MATERIALS： Six male SD rats of 6 months old and weighing （280＋20） g were used in this study. Chengzai pill was consisted of 22 Chinese medicines, such as Chinese Angelica, Eucommia Bark, Milkvetch Root, Barbary Wolfberry Fruit, Degelatined Deer-horn, Desertliving Cistanche, Eupolyphaga seu Steleophaga, Leech, Danshen Root and Himalayan Teasel Root, which were offered by Beijing Boran Pharmaceutical Co., Ltd. METHODS： The lipopolysaccharide and methylprednisolone were applied to prepare a rat model of steroid-induced osteonecrosis of femoral head. Six rats were divided into model group and Chengzai pill group at random with 3 rats in each group. The rats in the Chengzai pill group were administrated with methylprednisolone for the first time and then 1.5 g/kg Chengzai pill solution, once a day, totally for 6 weeks. The total RNA was extracted from femoral head in all rats 6 weeks later and then gene expression profiling was analyzed. MAIN OUTCOME MEASURES： Gene action pathway of cell adhesion molecules in gene expression profile. RESULTS： Compared with model rats, there were totally 8 downregulated genes of cell adhesion molecules which changed by a minimum of 1.5 folds in cell adhesion molecules pathway of Chengzai pill group, three rats were present with 633 genes （506 down-regulated and 127 up-regulated）, 883 genes （640 down-regulated and 243 up-regulated） and 593 genes （408 down-regulated and 185 up-regulated）, respectively. MAS software classification analysis showed 79 action pathways were involved in 297 genes. CONCLUSION： Following Chengzai pill administration, those downregulated genes recover the cognitive functions of macrophage and target cell in rats, also renew the normal survival condition of epithelial cells including vascular endothelial cells. Cell adhesion molecules action pathway returns to normal levels. This is the key to resume normal blood transport in femoral head of steroid-induced osteonecrosis.