龟甲提取物对骨髓间充质干细胞增殖过程中核受体的影响  

Effect of Extract of Carapax et Plastrum Testudinis on Nuclear Receptor in the Proliferation of Rat Mesenchymal Stem Cell in Vitro

作　　者：宋述财[1] 许华[2] 周健洪[1] 黎晖[1] 李伊为[1] 杜少辉[3] 陈东风[1] 

机构地区：[1]广州中医药大学,广州510405 [2]广州中医药大学第一附属医院,广州510405 [3]深圳市中医院,广东深圳518033

出　　处：《广州中医药大学学报》2006年第2期95-99,111,共6页Journal of Guangzhou University of Traditional Chinese Medicine

基　　金：国家自然科学基金项目(编号:30271677;30371837;30472272)广东省自然科学基金项目(编号:31483)广东省中医药管理局项目(编号:20513025)

摘　　要：【目的】观察龟甲(也称龟版或龟板)提取物对骨髓间充质干细胞(mesenchymal stem cells,MSCs)增殖过程中核受体 (nuclear receptor,NR)表达的影响。【方法】采用密度梯度法分离大鼠MSC进行培养,经5-溴脱氧尿嘧啶(Brdu)标记和 CD44免疫组化染色鉴定后,分别以高、中、低剂量的龟甲提取物(3 333、333．3、33．33μg／mL)加入到体外培养的MSC中, 与龟甲提取物作用12、24、72、120 h后,采用免疫组化法和免疫荧光细胞化学法检测MSC的视黄酸受体(retinoic acid receptor-α,RARα)、维生素D受体(vitamin D receptor,VDR)、雌激素受体(estrogen reeepror,ER)、糖皮质激素受体 (glucocorticoid receptor,GR)、甲状腺激素受体(thyroid hormone receptor-α,TRα)和过氧化物酶体增殖物活化受体(peroxisome proliferator activated receptor-δ,PPARδ)的表达。【结果】高、中、低剂量龟甲提取物组的MSC的RARα、VDR表达阳性细胞均高于对照组(P<0．05或P<0．01),且呈剂量依赖性;作用24 h后RARα表达达峰值,作用72 h后VDR表达达峰值。未检测到ER、GR、PPARδ、TRα阳性反应细胞。【结论】视黄酸受体α和维生素D受体可能为龟甲提取物促MSC增殖的药理靶点。[ Objective] To observe the effect of extract of Carapax et Plastrum Testudinis （ECPT） on nuclear receptor in the proliferation of rat mesenchymal stem cell （MSC） in vitro. [Methods] The rat MSC dissociated from bone marrow by density gradient method were euhured and identified by marking of bromodeoxyuridine （Brdu） and staining of CD44. Then different doses of ECPT （3333, 333.3 and 33.33 μg/mL） were respectively added into in-vitro cultured MSC for 12, 24, 72 and 120 hours. The expression of retinoic acid receptor-a （RARa）, vitamin D receptor （VDR）, estrogen receptor （ER）, glueocorticoid receptor （GR）, thyroid hormone receptor-α （TRα） and peroxisome proliferator-activated receptor δ （PPARδ） in MSC was detected by immunohistochemistry and immunofluorescence methods. [ Results] The number of RARα- and VDR-positive cells in ECPT groups was higher than that in the control group （ P 〈 0.05 or P 〈 0.01 ） and the effect was in a dose-dependant manner. RARa expression in MSC arrived at the pectk after 24-hour culture with ECPT, and VDR expression arrived at the peak after 72-hour culture. ER-, GR-, PPARδ- and TRα-positive cells were undetectable in ECPT groups. [ Conclusion] RARα and VDR probably are the pharmacological targets in ECPT promoting the proliferation of MSC.

关 键 词：龟甲提取物/药理学 骨髓间充质干细胞/药物作用 细胞增殖 核受体/病理学 细胞培养 大鼠 

分 类 号：R285.5[医药卫生—中药学]