[1]徐展望,李念虎.柚皮苷对体外培养骨髓间充质干细胞Runx-2和 Osterix表达及骨质疏松模型大鼠骨强度的影响[J].中医正骨,2013,25(12):7-10.
　Xu Zhanwang*,Li Nianhu.*.Effect of naringin on expression of Runx-2 and Osterix in bone marrow stem cell cultured in vitro and on bone strength for osteoporosis rat model[J].The Journal of Traditional Chinese Orthopedics and Traumatology,2013,25(12):7-10.
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柚皮苷对体外培养骨髓间充质干细胞Runx-2和 Osterix表达及骨质疏松模型大鼠骨强度的影响()
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《中医正骨》[ISSN:1001-6015/CN:41-1162/R]

卷:
第25卷
期数:
2013年12期
页码:
7-10
栏目:
骨质疏松症
出版日期:
2013-12-30

文章信息/Info

Title:
Effect of naringin on expression of Runx-2 and Osterix in bone marrow stem cell cultured in vitro and on bone strength for osteoporosis rat model
作者:
徐展望; 李念虎
山东中医药大学附属医院,山东 济南 250014
Author(s):
Xu Zhanwang*; Li Nianhu.*
The Affiliated Hospital of Shandong University of Traditional Chinese Medicine,Jinan 250014,Shangdong,China
关键词:
骨质疏松 柚皮苷 骨碎补 骨髓间充质干细胞 Runt相关转录因子2 Osterix 生物力学 动物实验
Keywords:
Osteoporosis;  Naringin;  Drynaria fortunei;  Bone marrow stem cell;  Runt-related transcription factor 2;  Osterix;  Biomechanics;  Animal experimentation
摘要:
目的:探讨柚皮苷对体外培养骨髓间充质干细胞Runx-2和Osterix表达及骨质疏松模型大鼠骨强度的作用。方法:采用髓腔冲洗离心法从雌性Lewis大鼠股骨获取骨髓间充质干细胞,将传代培养后的细胞分为5组。A组不进行干预,B组加入二甲基亚砜,C、D、E组分别加入浓度为1 μg·mL-1、10 μg·mL-1、100 μg·mL-1的柚皮苷。培养6 h后收集细胞采用RT-PCR法测定各组细胞Runx-2和Osterix的mRNA表达水平。将24只雌性SD大鼠的卵巢切除,3个月后将其随机分为4组,每组6只。Ⅰ组、Ⅱ组、Ⅲ组分别以60 mg·kg-1、300 mg·kg-1和1 500 mg·kg-1柚皮苷灌胃,Ⅳ组给予等体积的PBS灌胃,每天灌胃1次,持续2个月。药物干预结束后,测定大鼠股骨强度和胫骨骨小梁面积。结果:①Runx-2 mRNA和Osterix mRNA表达水平。B、C、D、E组大鼠骨髓间充质干细胞Runx-2 mRNA表达水平比较,差异有统计学意义[(1.10±0.02),(2.05±0.31),(2.76±0.14),(1.82±0.10),F=44.021,P=0.000]。进一步两两比较,C、D、E组Runx-2 mRNA表达水平高于B组(P=0.000,P=0.000,P=0.000); D组高于C组和E组(P=0.001,P=0.000); C组和E组比较,差异无统计学意义(P=0.153)。B、C、D、E组Osterix mRNA表达水平比较,差异有统计学意义[(1.02±0.10),(1.11±1.35),(3.24±0.30),(2.55±0.35),F=7.037,P=0.012]。进一步两两比较,D组和E组Osterix mRNA表达水平高于B组(P=0.031,P=0.005); C组和B组比较,差异无统计学意义(P=0.886); C组低于D组和E组(P=0.006,P=0.039); D组和E组比较,差异无统计学意义(P=0.267)。②股骨生物力学强度。4组骨质疏松模型大鼠股骨强度比较,差异有统计学意义[(773.36±9.21)N·mm-1,(805.66±16.00)N·mm-1,(766.70±38.96)N·mm-1,(707.46±15.88)N·mm-1,F=37.776,P=0.000]。进一步两两比较,Ⅰ、Ⅱ、Ⅲ组股骨强度均高于Ⅳ组(P=0.000,P=0.000,P=0.000); Ⅰ组和Ⅲ组比较,差异无统计学意义(P=0.509); Ⅱ组高于Ⅰ组和Ⅲ组(P=0.004,P=0.001)。③胫骨骨小梁面积。4组骨质疏松模型大鼠胫骨骨小梁面积比较,差异有统计学意义[(22.26±2.32),(25.10±2.18),(23.66±3.26),(15.63±2.00),F=14.168,P=0.000]。进一步两两比较,Ⅰ、Ⅱ、Ⅲ组胫骨骨小梁面积均大于Ⅳ组(P=0.001,P=0.000,P=0.000); Ⅰ组与Ⅱ组和Ⅲ组比较,差异均无统计学意义(P=0.090,P=0.387); Ⅱ组和Ⅲ组比较,差异无统计学意义(P=0.374)。结论:柚皮苷可促进体外培养的骨髓间充质干细胞中Runx-2和Osterix的表达,增强骨质疏松模型大鼠的骨强度,增大骨小梁面积。
Abstract:
Objective:To explore the effect of naringin on the expression of Runx-2 and Osterix in bone marrow stem cell(BMSC)cultured in vitro and on the bone strength for osteoporosis rat model.Methods:The BMSC were obtained from femur of femina Lewis rats by medullary cavity flushing and centrifugation,and the cells were divided into 5 groups after serial subcultivation.The cells in group A were not intervened and the cells in group B were placed in the culture fluids supplemented with dimethyl sulfoxide,while cells in other groups were placed in the culture fluids respectively supplemented with naringin with final concentration of 1 μg/mL(group C),10 μg/mL(group D)and 100 μg/mL(group E).The expression level of Runx-2 mRNA and Osterix mRNA were detected through RT-PCR after 6-hour culture.Twenty-four female SD rats were administrated with ovariectomy to creat models of osteoporosis and were randomly divided into 4 groups 3 months later,6 cases in each group.The rats in groupⅠ,Ⅱand Ⅲ were intragastric administrated with naringin with dose of 60 mg/kg(groupⅠ),300 mg/kg(groupⅡ)and 1 500 mg/kg(groupⅢ),while the rats in group Ⅳ were intragastric administrated with PBS, once a day for consecutive 2 months.After the drug intervention,the femur strength and the tibia trabecular bone area of the rats were measured.Results:There were statistical differences in Runx-2 mRNA expression level of BMSC among group B,C,D and E((1.10+/-0.02),(2.05+/-0.31),(2.76+/-0.14),(1.82+/-0.10),F=44.021,P=0.000).Further pairwise comparison showed that the Runx-2 mRNA expression level of group C,D and E were higher than that of group B(P=0.000,P=0.000,P=0.000),and group D surpassed group C and E(P=0.001,P=0.000),and there was no statistical difference between group C and group E(P=0.153).There were statistical differences in Osterix mRNA expression level of BMSC among group B,C,D and E((1.02+/-0.10),(1.11+/-1.35),(3.24+/-0.30),(2.55+/-0.35),F=7.037,P=0.012).Further pairwise comparison showed that the Osterix mRNA expression level of group D and group E were higher than that of group B(P=0.031,P=0.005),and there was no statistical difference between group C and group B(P=0.886),and the Osterix mRNA expression level of group C were lower than that of group D and group E(P=0.006,P=0.039),and there was no statistical difference between group D and group E(P=0.267).There were statistical differences in the femur strength among groupsⅠ,Ⅱ,ⅢandⅣ((773.36+/-9.21),(805.66+/-16.00),(766.70+/-38.96),(707.46+/-15.88)N/mm,F=37.776,P=0.000).Further pairwise comparison showed that the femur strength of groupⅠ,Ⅱand Ⅲ were higher than that of group Ⅳ(P=0.000,P=0.000,P=0.000),and there was no statistical difference between groupⅠand group Ⅲ(P=0.509),and the femur strength of group Ⅱ were higher than that of groupⅠand group Ⅲ(P=0.004,P=0.001).There were statistical differences in the tibial bone trabecula area among groupⅠ,Ⅱ,Ⅲ and Ⅳ((22.26+/-2.32),(25.10+/-2.18),(23.66+/-3.26),(15.63+/-2.00),F=14.168,P=0.000).Further pairwise comparison showed that the tibial bone trabecula area of groupⅠ,Ⅱand Ⅲ were larger than that of group Ⅳ(P=0.001,P=0.000,P=0.000),and there were no statistical differences among groupⅠ,Ⅱand Ⅲ(P=0.090,P=0.387),and there was no statistical difference between groupⅡand group Ⅲ(P=0.374).Conclusion:Naringin can promote the expression of Runx-2 and Osterix in BMSC cultured in vitro,and it can increase bone strength and bone trabecula area of osteoporosis rat.

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备注/Memo

备注/Memo:
2013-09-15收稿 2013-11-01修回
基金项目:山东省自然科学基金面上项目(Y2008C147)
通讯作者:徐展望 E-mail:xzw6001@163.com
更新日期/Last Update: 2013-12-30