基于蛋白质组学技术的鹿茸治疗大鼠OA的分子靶点筛选研究

Identification of Molecular Network Target of Antler on OA Rat Using Proteomics Technology

目的从信号转导系统与受体学说角度,研究鹿茸对大鼠各脏腑cAMP/cGMP、TGF-β水平的影响,探讨其与归经的相关性。方法选择3月龄健康雌性SD大鼠24只,体质量(200±20)g,随机分为4组(鹿茸低、中、高剂量组及空白对照组)。常规喂养1周后,除正常组外采用经典Hulth方法制备骨关节炎疾病模型,造模6周后模型大鼠按分组不同给予鹿茸低、中、高剂量灌喂2周,空白对照组予等量生理盐水灌喂,通过酶联免疫吸附实验和免疫组织化学实验检测鹿茸灌喂后组内脏组织cAMP/cGMP及TGF-β受体的表达水平,通过比较各内脏组织cAMP/cGMP及TGF-β受体相对表达量变化程度,分析鹿茸对各内脏组织的作用趋势。结果鹿茸低剂量组脾脏cAMP/cGMP水平明显升高,差异有统计学意义(P<0.05);鹿茸中剂量组肾脏、心脏cAMP/cGMP水平明显升高,差异有统计学意义(P<0.05);鹿茸高剂量组肝脏、卵巢cAMP/cGMP水平明显升高,差异有统计学意义(P<0.05);cAMP/cGMP相对表达量由大到小排序为:肾、肝、卵巢、脾、心、脑、胃、肺、肠。鹿茸低、中剂量组肝、肾TGF-β受体表达水平明显升高,差异有统计学意义(P<0.05);鹿茸低剂量组脾、胃,鹿茸中剂量组脾,鹿茸高剂量组肠、胃TGF-β受体表达水平明显下降,差异有统计学意义(P<0.05);TGF-β受体相对表达量由大到小排序为:肾、肝、卵巢、肺、心、脑、肠、胃、脾。结论鹿茸对大鼠信号转导系统水平变化及受体表达水平的影响作用以肾脏、肝脏为主,与传统鹿茸归经理论相符。

Objective To observe the effects of pilose antler on the levels of cAMP/cGMP and TGF-β in viscus of rats from the perspective of signal transduction system and receptor theory and explore the channel tropism of pilose antler. Methods Twenty four 3-month old female healthy SD rats weighted(200±20)g were recruited and routinely fed for 1 week. They were randomly divided into 4 groups:the PA low dose group,the PA medium dose group,the PA high dose group,the normal control group,6 in each group. The model was prepared using classic Hulth method except the normal control group. After 6-week modeling,the model rats were fed with low,medium and high dose for 2 weeks according to different groups. The normal control group was fed with the same amount of normal saline. The levels of cAMP/cGMP and TGF-β receptor in visceral tissues were detected by enzyme-linked immunosorbent assay and immunohistochemistry. The trend of effect of PA on visceral tissues was analyzed by comparing the changes of the relative expressions of cAMP/cGMP and TGF-β receptor in visceral tissues. Results The levels of cAMP/cGMP in spleen of the low dose PA group increased significantly(P<0.05). The levels of cAMP/cGMP in kidney and heart of the medium dose PA group increased significantly(P<0.05) while the levels of cAMP/cGMP in liver and ovary of the high dose PA group increased significantly(P<0.05). And the rank of the relative expression of cAMP/cGMP from large to small order were kidney,liver,ovary,spleen,heart,brain,stomach,lung and intestine. The levels of TGF-β receptor in kidney and liver of the low and medium dose PA groups increased significantly(P<0.05). The levels of TGF-β receptor in spleen and stomach of the low and medium dose PA groups,in spleen of the medium dose PA group and in intestine and stomach of the high dose PA group decreased significantly(P<0.05). The rank of the relative expression of TGF-β receptor from large to small order were kidney,liver,ovary,lung,heart,brain,intestine,stomach and spleen. Conclusions The effect of pilose antler on cAMP/cGMP and TGF-β is mostly in kidney and liver,and this view conforms to the traditional channel tropism of pilose antler in traditional Chinese medicine theory.