3种补肾中药有效成分对去卵巢骨质疏松大鼠骨髓间充质干细胞的调控作用  

Regulation and control of three active components of TCM effective in reinforcing kidney on ovariotomy-induced osteoporosis rat's bone marrow stem cells revealed by gene microarray

作　　者：卞琴[1,2,3] 刘书芬[3] 黄建华[1,2] 杨铸[3] 宁友[1,2] 赵永见[3] 王拥军[3] 沈自尹[1,2] 

机构地区：[1]复旦大学中西医结合研究所 [2]复旦大学华山医院,上海200040 [3]上海中医药大学脊柱病研究所上海中医药大学龙华医院,上海200032

出　　处：《中华中医药杂志》2011年第5期889-893,共5页China Journal of Traditional Chinese Medicine and Pharmacy

基　　金：国家重点基础研究发展计划(973计划)(No.2010CB530402;2010CB530400);国家自然科学基金青年项目(No.81001526)~~

摘　　要：目的:观察3种补肾代表中药淫羊藿、补骨脂、女贞子各自有效成分淫羊藿苷、补骨脂素、齐墩果酸对去卵巢3个月大鼠骨髓间充质干细胞(BMSCs)的调控作用。方法:40只Sprague Dawley(SD)雌性大鼠随机分为正常对照组、模型组、淫羊藿苷组、补骨脂素组、齐墩果酸组(后3组合称治疗组)。模型组和治疗组切除双侧卵巢,正常对照组切除卵巢周围部分脂肪组织。治疗组于造模第2d开始按20mg/kg给予对应药物每日1次灌胃,连续12周。造模后第4周开始称量体质量,每周1次。造模后12周大鼠处死,取第4腰椎进行micro-CT扫描,评价骨量改变情况。采用全骨髓贴壁培养法培养原代BMSCs,于取材后第7d干细胞基因芯片技术检测mRNA表达。结果:造模后大鼠体质量增加,补骨脂素能减少增加幅度,而淫羊藿苷和齐墩果酸没有明显作用。Micro-CT显示,去卵巢12周后,模型组与正常对照组之间,骨形态计量学参数骨体积与组织体积之比BV/TV,骨小梁数目Tb.N,骨小梁厚度Tb.Th明显减少,骨小梁空间距离Tb.Sp明显增加。淫羊藿苷和齐墩果酸显著增加Tb.Th,对其他参数也有改善的趋势。补骨脂素有改善的趋势,但没有统计学差异。大鼠全基因表达谱显示在被检测的26 420个基因中,模型组比正常对照组下调2倍的基因有4 193个,约占15.87%,淫羊藿苷、补骨脂素和齐墩果酸比模型组上调2倍的基因分别有3 226,1 701和3 232个。模型组比正常对照组上调2倍的基因有5 015个,约占18.98%,3个有效成分比模型组下调2倍的基因分别有3 549,1 660和3 545个。基因功能分类分析显示3个有效成分共同改变的基因涉及的功能包括:细胞成分中的初级内体、分子功能相关的磷酸跨膜转运蛋白活性、GDP结合、内肽酶活性、酶调节剂活性、谷氨酸受体结合、转移酶活性、5-羟色胺受体活性、血清素结合;细胞外基质、跨膜受体蛋白酪氨酸激酶活性、�Objective：To observe the effects of icariin,Psoralen and oleanolic acid,three active components of epimedium,psoralea corylifolia and glossy privet fruit respectively,on ovariectomized-induced rats＇ bone marrow stem cells.Methods：Forty SD femal rats were randomly divided into normal control group,untreated group,icariin group,Psoralen group and oleanolic acid group（the last three called treatment groups）.The rats in untreated group and treatment groups were ovariotomized while some of adipose tissue around the ovary was cut off in the normal control group.Rats in the treatment groups were intragastrically administered with icarrin,or Psoralen,or oleanolic acid at dose of 20mg/（kg？d） for 12 weeks.The body weight was obtained once a week on the beginning of the 4th week after the model creation.All rats were sacrificed and the fourth lumbar vertebras were harvested for micro-CT scanning to evaluate bone mass.BMSCs were obtained ex vivo by the methods of bone marrow adherent culture.The mRNAs of BMSCs were detected using Gene Chip technique on the 7th day of culture.Results：There was an increase in body weight of rats in the untreated group compared with that in the normal group.Psoralen could decrease this increasing extent while icariin and oleanolic acid could not.Micro-CT showed an increase in morphometry such as BV/TV,Tb.N and Tb.Th,and a decrease in Tb.Sp in lumbar vertebral morphometry between untreated and normal control groups.The value of Tb.Th was significantly increased.In microarray（26420 genes）,there were 4193 genes（15.87%） downregulated in the untreated groups compared with those in the normal control groups.Among them,there were 3226,1701,3232 genes were upregulated by icariin,psoralen and oleanolic acid.On the other hand,there were 5015 genes（18.98%） upregulated in the untreated groups compared with those in the normal control groups.Among them,there were 3549,1660,3545 genes were downregulated by icariin,psoralen and oleanolic acid.GO ananlysis showed these genes funct

关 键 词：干细胞基因芯片 补肾药 淫羊藿苷 补骨脂素 齐墩果酸 间质干细胞 基因表达谱 大鼠 

分 类 号：R285.5[医药卫生—中药学]