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中国人民解放军总医院老年心血管病研究所
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中华老年多器官疾病杂志编辑委员会
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创刊人 王士雯
总编辑 范利
副总编辑 陈韵岱
执行主编 叶大训
编辑部主任 王雪萍
ISSN 1671-5403
CN 11-4786
创刊时间 2002年
出版周期 月刊
邮发代号 82-408
友情链接
张泽华,霍建忠,郭耀,季兴华.甲状旁腺素联合骨髓间充质干细胞对骨质疏松大鼠股骨颈骨密度相关指标的影响[J].中华老年多器官疾病杂志,2019,18(1):45~50
甲状旁腺素联合骨髓间充质干细胞对骨质疏松大鼠股骨颈骨密度相关指标的影响
Effect of parathyroid hormone combined with BMSCs transplantation on femur bone density in osteoporotic rats
投稿时间:2018-09-19  修订日期:2018-11-27
DOI:10.11915/j.issn.1671-5403.2019.01.010
中文关键词:  骨髓间充质干细胞;甲状旁腺激素;骨质疏松
英文关键词:bone marrow mesenchymal stem cells; parathyroid hormone; osteoporosis
基金项目:山西省科技厅科技攻关项目(20150313012-1)
作者单位E-mail
张泽华 山西医科大学附属大医院骨科,太原 030001  
霍建忠 山西医科大学附属大医院骨科,太原 030001 huojianzhong@126.com 
郭耀 山西医科大学附属大医院骨科,太原 030001  
季兴华 山西医科大学附属大医院骨科,太原 030001  
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中文摘要:
      目的 探索骨髓间充质干细胞(BMSCs)联合甲状旁腺激素(PTH)与二者单独使用对骨质疏松性大鼠骨密度相关指标的影响。方法 将50只SD雌性大鼠分为A、B、C、D、E组,每组10只,B、C、D、E组切除双侧卵巢,A组大鼠仅切除卵巢周围同等大小的脂肪组织。术后12周各组抽取3只大鼠,检测大鼠的雌激素水平及骨密度相关指标骨小梁数量(Tb.N)、骨小梁厚度(Tb.Th)、骨小梁分离度(Tb.Sp)和骨体积分数(BV/TV)。脱颈法处死SD健康雄性大鼠取骨髓进行BMSCs细胞培养,流式细胞技术单标法检测BMSCs是否培养成功。术后12周,各组剩余7只大鼠。其中A和B组不做后期干预处理;C组按60 μg/kg剂量背部皮下注射Rat PTH 1-34,隔日1次,连续12周;D组显露双侧股骨颈,注入浓度为5×108/ml的BMSCs;E组在D组的基础上在皮下注射PTH,注射浓度、频率与C组一致。所有大鼠均在干预12周后处死,Micro-CT测量5组大鼠股骨颈Tb.N、Tb.Th、Tb.Sp和BV/TV,观察成骨情况。使用SPSS 20.0软件进行统计分析。组间比较采用单因素方差分析,有统计学意义再进行LSD两两比较。结果 术后12周,与A组比较,B、C、D及E组大鼠雌激素水平、Tb.N、Tb.Th及BV/TV显著下降,Tb.Sp明显升高,差异有统计学意义(P<0.05),但B、C、D及E组各组间上述指标差异不明显(P>0.05),可证明B、C、D及E组大鼠为骨质疏松模型。流式细胞技术单标法显示,P3代BMSCs表达CD29(99.86%)和CD90(99.73%),几乎不表达CD34(6.48%)和CD45(0.94%),证明BMSCs培养成功。干预12周后,Micro-CT数据显示,单独使用PTH或BMSCs及联合使用均可改善骨微观结构,但均未达到A组水平。与B组比较,C、D、E组的Tb.N、Tb.Th及BV/TV显著增加,Tb.Sp显著下降,差异有统计学意义(P<0.05);与C或D组比较,E组Tb.N、Tb.Th及BV/TV显著增加,Tb.Sp显著下降,差异有统计学意义(P<0.05)。结论 BMSCs联合PTH治疗局部骨质疏松的效果优于单独使用治疗的效果。
英文摘要:
      Objective To explore the effects of bone marrow mesenchymal stem cells (BMSCs) transplantation or parathyroid hormone (PTH) alone or combined together on related indices of bone density in osteoporotic rats. Methods Fifty female SD rats were divided into groups A, B, C, D and E, 10 animals in each group. Bilateral ovaries were removed in the rats from groups B, C, D and E, while for those from group A, adipose tissue around the ovaries was only removed. In 12 weeks after operation, 3 rats were selected from each group, and then the estrogen level and bone density related indices such as trabecular number (Tb.N), trabecular thickness (Tb.Th), trabecular separation (Tb.Sp) and bone volume to tissue volume (BV/TV) were measured. SD healthy male rats were sacrificed to isolate and culture BMSCs, and the obtained cells were further identified with single labeling flow cytometry. At 12 weeks after operation, except those from groups A and B getting no intervention, subcutaneous injection of 60 μg/kg rat PTH 1-34 on the back (once every other day, for 12 consecutive weeks) was given to group C and the injection of 5×108/ml BMSCs on the bilateral femoral neck were given to the group D, and group E were given the same intervention as group C on the basis of group D. The rest 7 rats in five groups were sacrificed in 12 weeks after intervention. Tb.N, Tb.Th, Tb.Sp and BV/TV of femoral neck were measured by micro-CT scanning. SPSS statistics 20.0 was used for statistical analysis. One-way ANOVA was employed for comparisonamong groups, and LSD test was applied for those with statistical significance. Results Compared with group A, the estrogen level, Tb.N, Tb.Th and BV/TV in groups B, C, D and E were decreased significantly, and Tb.Sp was increased significantly (P<0.05), but there was no significant differences among groups B, C, D and E (P>0.05), and the rats in group B, C, D and E were identified as osteoporotic rats. Single-labeling flow cytometry showed that CD29 (99.86%) and CD90 (99.73%) were expressed in the third passaged (P3) BMSCs, while CD34 (6.48%) and CD45 (0.94%) were rarely expressed, which proved that BMSCs were successfully cultured. After 12 weeks of intervention, micro-CT scanning showed that PTH or BMSCs alone, or combination improved bone microstructure, but they did not reach the level of group A. Compared with group B, Tb.N, Tb.Th and BV/TV in groups C, D and E were increased significantly, while Tb.Sp was decreased significantly (P<0.05). Compared with groups C or D, Tb.N, Tb.Th and BV/TV in group E were increased significantly and Tb.Sp was decreased significantly (P<0.05). Conclusion The effect of BMSCs combined with PTH in the treatment of local osteoporosis is better than that of single use.
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