阳之韵,兰波,刘亭,李勇军,陈孝
YANG Zhiyun,LAN Bo,LING Ting,LI Yongjun,CHEN Xiao

• 1:中山大学药学院
• 2:贵州省人民医院
• 3:贵州医科大学药物制剂重点实验室
• 4:中山大学附属第一医院

摘要(Abstract)：

目的:研究杜仲中各成分对MC3T3-E1 Subclone 14成骨细胞增殖、成骨细胞保护素(DPG)与核因子κB受体活化因子配体(RANKL)比值的影响。方法:对杜仲皮进行组分提取,得到A组分(水洗脱液)、b组分(20%乙醇洗脱液)和C组分(40%乙醇洗脱液);对照组MC3T3-E1 Subclone 14成骨细胞加入DMEM/F-2培养基(Hyclon)培养,给药组分别加入浓度为200、100、50、25和12.5 mg/L含A、B、C组分的培养基培养;采用MTT法检测MC3T3-E1 Subclone 14成骨细胞增殖情况;ELISA法检测MC3T3-E1 Subclone 14成骨细胞OPG及RANKL活性,紫外分光光度法检测MC3T3-E1 Subclone 14成骨细胞碱性磷酸酶(ALP)活性。结果:与对照组比较,给药24 h后,100和200 mg/L C组分给药组的OD值明显增高(P<0.05);与对照组比较,给药48 h后,200 mg/LC组ALP的活性以及OPG与RANKL比值明显增高,差异具有统计学意义(P<0.05)。结论:杜仲40%乙醇提取组分能明显上调OPG与RANKL比值,并能促进MC3T3-E1 Subclone 14成骨细胞的增殖。
Objective: To screen effective components of Eucommia ulmoides for studying their effents on the proliferation of osteoblasts and ratio of OPG/RANKL in MC3T3-E1 Subclone 14 osteoblasts. Methods: Extracting component of Eucommia,and dividing into A group(water washing liquid),B group(20% ethanol washing liquid) and C group(40% ethanol washing liquid); control group MC3T3-E1 Subclone 14 osteoblasts was added with DMEM/F-2 culture medium(Hyclon),other groups were added with A,B,C components at the concentration of 200,100,50,25 and 12. 5 mg/L;MTT detection method to detect cell proliferation; the activity of OPG and RANKL are detected by ELISA; UV spectrophotometry was adopted to detect the ALP activity of MC3T3-E1 Subclone 14 osteoblasts. Results: Comparing with control group,24 hr after medication,OD value of 100 and 200 mg/L C group significantly increased(P < 0. 05); comparing with control group,48 hr after medication,ALP activity and OPG/RANKL ratio of 200 mg/L C group significantly increased,differences were statistically significant(P < 0. 05). Conclusion: 40% ethanol Eucommia ulmoides Oliv extract can up regulate the OPG/RANKL ratio,and enhance osteoblast proliferation.

关键词(KeyWords)： 杜仲;成骨细胞;骨保护素;碱性磷酸酶
Eucommia ulmoides Oliv;osteoblasts;osteoprote-gerin;alkaline phosphatase

Abstract:

Keywords:

基金项目(Foundation): 贵州省科技重大专项[黔科合重大专项字(2012)6009号];; 贵州省中药现代化专项项目[黔科合中药字(2013)5062号]

作者(Author): 阳之韵,兰波,刘亭,李勇军,陈孝
YANG Zhiyun,LAN Bo,LING Ting,LI Yongjun,CHEN Xiao

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