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| CKIP-1过表达对大鼠体外破骨细胞增殖影响的研究 |
| The effect of CKIP-1 overexpression on the proliferation of osteoclasts in vitro |
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| DOI：10.3969/j.issn.1006-7108.2018.10.005 |
| 中文关键词: CKIP-1 破骨细胞 大鼠 细胞增殖 动物实验 骨质疏松 |
| 英文关键词:CKIP-1 osteoclasts rats cell proliferation animal experimentation osteoporosis |
| 基金项目:国家自然科学基金项目（81373878） |
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| 中文摘要: |
| 目的 分离培养大鼠破骨细胞，构建CKIP-1过表达慢病毒，观察被CKIP-1过表达慢病毒感染的体外大鼠破骨细胞的细胞增殖情况，探讨CKIP-1基因对体外大鼠破骨细胞增殖的影响。方法 分离、培养SD大鼠原代破骨细胞，用抗酒石酸酸性磷酸酶染色法鉴定。构建CKIP-1过表达载体，完成慢病毒包装。将破骨细胞分为A组(破骨细胞空白对照组)、B组(破骨细胞+空载病毒组)、C组(破骨细胞+CKIP-1过表达病毒组)，分别进行对应处理，qRT-PCR检测CKIP-1基因表达效果，CCK-8试剂盒检验破骨细胞的相对数量。结果 成功鉴定大鼠原代破骨细胞分离培养。qRT-PCR检测破骨细胞中CKIP-1的mRNA水平结果显示：相比A组和C组，在感染CKIP-1病毒的C组，CKIP-1的基因水平有较高表达（P＜0.01），CKIP-1过表达载体过表达效果明显，载体构建成功。CCK-8试剂盒检测各组细胞增殖的结果显示：相比A组和C组，感染CKIP-1病毒的C组的细胞增殖比例显著升高（P＜0.01）。结论 成功分离培养出大鼠破骨细胞。成功构建CKIP-1过表达慢病毒，并感染体外大鼠破骨细胞。CKIP-1的过表达具有促进体外大鼠破骨细胞增殖的功能。 |
| 英文摘要: |
| Objective To isolate and culture rat osteoclasts and construct CKIP-1 overexpression lentivirus, to observe the proliferation of rat osteoclasts infected by CKIP-1 overexpressing lentivirus in vitro, and to investigate the effect of CKIP-1 gene on rat osteoclast proliferation in vitro. Methods Rat primary osteoclasts were isolated and cultured and identified by tartrate acid phosphatase staining. The CKIP-1 overexpression vector was constructed to complete the lentivirus packaging. The osteoclasts were grouped into: Group A without special treatment, Group B with no-load virus infection, and Group C with CKIP-1 overexpression lentivirus infection. Corresponding treatments were given, qRT-PCR was used to detect CKIP-1 gene expression effect, and the CCK -8 kit was used to test the relative amount of osteoclasts. Results The rat primary osteoclasts were isolated and cultured successfully. qRT-PCR detection of CKIP-1 mRNA in osteoclasts showed that CKIP-1 gene was highly expressed in Group C that was infected with the CKIP-1 virus compared with Groups A and C (P<0.01). The overexpression of CKIP-1 overexpression vector was effective and the vector construction was successful. The results of CCK-8 kit detection of cell proliferation in each group showed that the proportion of cell proliferation in Group C that was infected with CKIP-1 virus was significantly higher than that in Groups A and C (P<0.01). Conclusion The results showed that rat osteoclasts were successfully isolated and cultured, and CKIP-1 overexpressing lentivirus was successfully constructed and infected rat osteoclasts in vitro. Overexpression of CKIP-1 could promote the proliferation of rat osteoclasts in vitro. |
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