低剂量X线照射对骨髓间充质干细胞增殖和分化的影响

发布时间：2018-03-20 05:33

  本文选题：间充质干细胞　切入点：低剂量照射　出处：《苏州大学》2012年硕士论文　论文类型：学位论文

【摘要】：目的：研究低剂量X线照射对离体培养的骨髓间充质干细胞增殖和分化的影响，探讨低剂量照射促进大鼠骨折后骨痂矿化的细胞机制。 方法：1.由大鼠骨髓中提取间充质干细胞（BMSCs），分别以0Gy、25mGy、50mGy、75mGy、100mGy、1000mGy不同剂量的X线照射，以CCK-8法检测照射6h、12h、24h、48h、72h后细胞的增殖情况，以流式细胞技术检测照射后对细胞周期的影响。2.如前述剂量照射后，诱导BMSCs向成骨细胞方向分化，评估分化的成骨细胞的成骨潜能。以Vonkossa染色法计数矿化结节的形成数目并评估其成骨能力。以RT-PCR法检测成骨相关基因OPG、COL-1、BGP的表达情况。 结果：1.CCK-8法检测结果表明75mGy低剂量照射后，可明显刺激BMSCs的增殖，与假照射组相比有统计学意义（P＜0.05）。而1000mGy高剂量照射则会抑制BMSCs增殖（P＜0.05）。流式细胞技术检测表明，，低剂量X线照射可以促进BMSCs由G1期进入S期，与假照射组相比具有统计学意义（P＜0.05）。2.向成骨细胞诱导分化后，低剂量照射组矿化结节数目较假照射组明显增多，以75mGy与100mGy组最为明显（P＜0.05）。RT-PCR结果显示OPG、COL-1、BGP经低剂量X线照射后表达量均有明显增高，以以75mGy与100mGy组最为明显（P＜0.05），而1000mGy高剂量照射则会抑制相关成骨基因的表达。 结论：1.低剂量X线照射可有效促进BMSCs的增殖，而高剂量照射则有抑制作用。2.低剂量X线照射可增强BMSCs的分化能力，照射后BMSCs成骨潜能明显增强，而高剂量照射起到相反的作用。
[Abstract]:Aim: to study the effects of low dose X-ray irradiation on the proliferation and differentiation of bone marrow mesenchymal stem cells (MSCs) cultured in vitro and to explore the cellular mechanism of bone callus mineralization induced by low dose irradiation. Methods 1. Bone marrow mesenchymal stem cells (BMSCs) were extracted from rat bone marrow and irradiated with different doses of 0 Gy 25 mGy 50 mGy 75 mGy 100 mGy respectively. The proliferation of cells was detected by CCK-8 method after irradiation for 6 h 12 h or 24 h or 48 h for 72 h. Flow cytometry was used to detect the effect of irradiation on cell cycle. (2) BMSCs was induced to differentiate into osteoblasts, The osteogenic potential of differentiated osteoblasts was evaluated. The number of mineralized nodules was counted and the osteogenic ability was evaluated by Vonkossa staining. The expression of osteoblastic associated gene OPGG-COL-1 was detected by RT-PCR method. Results 1. The results of CCK-8 assay showed that 75 mGy could significantly stimulate the proliferation of BMSCs after low dose irradiation, which was significantly higher than that of sham irradiation group (P < 0.05), while high dose irradiation of 1000mGy could inhibit the proliferation of BMSCs (P < 0.05). Flow cytometry showed that the proliferation of BMSCs was inhibited by high dose irradiation (P < 0.05). Low dose X-ray irradiation could promote BMSCs from G1 phase to S phase. Compared with false irradiation group, the number of mineralized nodules in low dose irradiation group was significantly higher than that in false irradiation group (P < 0.05, P < 0.05, P < 0.05, P < 0.05, P < 0.05, P < 0.05, P < 0.05, P < 0.05, P < 0.05). The results of RT-PCR showed that the expression of BGP in 75mGy and 100mGy groups was significantly increased after low dose X-ray irradiation, especially in 75mGy and 100mGy groups (P < 0.05), while the expression of osteogenic genes was inhibited by high dose radiation of 1000mGy. Conclusion low dose X ray irradiation can effectively promote the proliferation of BMSCs, while high dose X ray irradiation can inhibit the proliferation of BMSCs. Low dose X ray irradiation can enhance the differentiation ability of BMSCs, and the osteogenic potential of BMSCs is obviously enhanced after irradiation. High doses of radiation have the opposite effect.
【学位授予单位】：苏州大学
【学位级别】：硕士
【学位授予年份】：2012
【分类号】：R329

【参考文献】

相关期刊论文 前4条

1 高建军,高林峰,金慰芳,王洪复,魏道林,李凌波,高艳虹,周轶;γ射线对体外培养骨细胞的作用观察[J];辐射研究与辐射工艺学报;2002年01期

2 马杰;陈斌;王宏兰;李静;史明霞;李炳宗;胡建立;赵春华;;全身照射引起间充质干/祖细胞池缩小及成骨潜能改变[J];中国实验血液学杂志;2007年02期

3 刘树铮;低剂量辐射兴奋效应发生机理若干问题的探讨[J];中华放射医学与防护杂志;2003年06期

4 王献理，苏士杰，尹洪淑，冯乐平，于开君;低剂量辐射对小鼠种植肿瘤及其放疗的影响[J];中华放射医学与防护杂志;1996年03期

本文编号：1637761