补肾、健脾和补肾健脾3方对尾部悬吊大鼠骨髓间充质干细胞成骨分化的影响  

Effects on Osteogenic Differentiation of Bone Mesenchymal Stem Cells in Tail Suspension Model Rats by Kidney-tonifying, Spleen-invigorating and Kidney-tonifying Combined with Spleen-invigorating Prescriptions

作　　者：张林[1] 谢鑫[1] 张红梅[1] 卢健[1] 陈文娜[1] 王俊岩[1] 李环宇[1] 

机构地区：[1]辽宁中医药大学基础医学院,沈阳110847

出　　处：《世界科学技术-中医药现代化》2015年第1期83-88,共6页Modernization of Traditional Chinese Medicine and Materia Medica-World Science and Technology

基　　金：国家自然科学基金委青年基金项目(81001493):基于脾肾相关理论防治模拟航天失重条件下骨丢失的多靶点作用研究;负责人:张林

摘　　要：目的:观察补肾、健脾和补肾健脾3方对尾部悬吊模拟失重大鼠骨髓间充质干细胞成骨分化能力的影响。方法:大鼠随机分为正常组、模型组、补肾组、健脾组和补肾健脾组共5组。后4组大鼠头低位-30°尾部悬吊连续21天,补肾、健脾和补肾健脾组大鼠从实验第1天开始分别给予补肾、健脾和补肾健脾方灌胃,其余各组大鼠灌服等容积的生理盐水。实验第22天处死各组大鼠,全骨髓贴壁法体外分离培养大鼠骨髓间充质干细胞(BMSCs),取第3代细胞进行成骨分化诱导实验,成骨分化诱导第4、7、10天采用p-NPP法检测碱性磷酸酶(ALP)活性,成骨分化诱导第7、14、21天采用ELISA法检测细胞上清液中骨钙素(OC)含量,成骨分化诱导第21天茜素红染色法检测钙结节形成情况。结果:与正常组比较,模型组大鼠BMSCs经成骨诱导4、7、10天ALP活性均显著降低(P<0.01),经成骨诱导7、14、21天细胞上清OC含量显著降低(P<0.01),经成骨诱导21天后钙结节形成显著减少(P<0.01);与模型组比较,补肾组和补肾健脾组大鼠BMSCs经成骨诱导4、7、10天ALP活性均明显升高(P<0.05或P<0.01),健脾组大鼠BMSCs经成骨诱导7、10天ALP活性均明显升高(P<0.05),补肾、健脾和补肾健脾组大鼠BMSCs经成骨诱导7、14、21天细胞上清OC含量明显升高(P<0.05或P<0.01),经成骨诱导21天后钙结节形成显著增多(P<0.01);与补肾组比较,健脾组大鼠BMSCs经成骨诱导7天ALP活性降低、经成骨诱导21天细胞上清OC含量明显降低(P<0.05),而补肾健脾组明显升高(P<0.05),经成骨诱导21天后补肾健脾组钙结节形成明显增多(P<0.05)。结论:补肾、健脾和补肾健脾3方具有促进尾部悬吊模拟失重大鼠骨髓间充质干细胞成骨分化的作用,以补肾健脾方作用为优。This study was aimed to observe the action of kidney-tonifying （KT）, spleen-invigorating （SI）, and kidney- tonifying combined with spleen-invigorating （KTSI） prescriptions on osteogenic differentiation potential of bone marrow mesenchymal stem cells （BMSCs） in tail suspension simulated weightlessness model rats. Rats were randomly divided into 5 groups, which were the control （C） group, tail suspension （S） group, KT group, SI group, and KTSI group. Rats in the S, KT, SI and KTSI groups were suspended by head down tilting -30° for 21 days. On the 1^st day of the experiment, oral administration of herbal decoction was given to rats of KT, SI and KTSI groups, respectively. Oral administration of equivalent amount of saline was given to rats in other groups. Rats of all groups were sacrificed on the 22^nd day of the experiment. BMSCs were isolated and cultured by adherence method on the 22^nd day. The third passage cells were induced by osteogenic differentiation solution. The activity of ALP was detected by the method of p-NPP on the 4^th, 7^th and 10^th day of osteogenic differentiation. The content of osteocalcin （OC） in culture supernatant was detected by ELISA on the 7^th, 14^th and 21^st day of osteogenic differentiation. The test of calcium nodules was made by alizarin red staining on the 21^st day of osteogenic differentiation. The results showed that compared with the C group, ALP activity was decreased significantly on the 4^th, Th and 10^th day of osteogenic differentiation in the S group （P〈0.01）; and the content of OC was obviously reduced on the 7^th, 14^th and 21^st day of osteogenic differentiation （P〈0.01）. The calcium nodules were reduced on the 21st day of osteogenic differentiation （P〈0.01）. Compared with the S group, the ALP activity was obviously increased on the 4th, 7th and ]0th day of osteogenic differentiation in the KT and KTSI groups （P〈0.05, or P〈0.01）. The ALP activity was obviously increased on the 7^th and 10^th day of osteogenic d

关 键 词：补肾健脾 尾部悬吊 模拟失重 成骨分化 骨髓间充质干细胞 

分 类 号：R285.5[医药卫生—中药学]