摘 要 目的：探讨Rho激酶抑制剂Y-27632在雷奈酸锶（Sr）促进大鼠骨髓间充质干细胞（rBMSCs）向成骨细胞分化中的作用。方法：体外分离SD大鼠原代骨髓间充质干细胞，并采用贴壁培养法培养细胞并传至第3代。接种细胞后依据实验目的进行分组，空白、加Y抑制剂、加Sr及Sr+Y抑制剂共4组分别在第1、4和7天用CCK-8试剂盒检测细胞增殖情况，用BCA蛋白浓度试剂盒测定各组细胞的蛋白总量进而用ALP试剂盒通过酶标仪检测细胞碱性磷酸酶活性高低，最后用RT-PCR法检测各组细胞特异性转录因子Runx-2及下游成骨蛋白OCN、OPN等促进成骨分化的基因的表达量情况。 结果：（1）Rho激酶抑制剂Y-27632作用于rBMSCs后，细胞生长加快，增殖情况高于空白组；Rho激酶抑制剂Y-27632在Sr中作用于rBMSCs增殖情况也高于单独加Sr一组。（2）Rho激酶抑制剂Y-27632相对于空白组ALP活性增加幅度不大；Rho激酶抑制剂Y-27632在Sr中作用于rBMSCs ALP活性相比于单独加Sr一组显著增加。（3）Rho激酶抑制剂Y-27632相对于空白组基因表达量增加显著；Rho激酶抑制剂Y-27632在Sr中作用于rBMSCs基因表达量相对于单独加Sr一组增加极显著。结论: Rho激酶抑制剂Y -27632在雷奈酸锶促进大鼠骨髓间充质干细胞向成骨细胞分化中主要起促进作用。69167

该论文有图8幅，表1个，参考文献20篇。

毕业论文关键词 : 骨髓间充质干细胞  Rho激酶抑制剂  信号通路  增殖  基因表达

Effects of Rho kinase inhibitor Y-27632 in strontium ranelate –induced osteogenic differentiation of rat bone marrow mesenchymal stem cells

ABSTRACT

   Aim: To study the role of Rho kinase inhibitor Y-27632 in osteogenic differentiation of rat bone marrow mesenchymal stem cells induced by strontium ranelate. Methods: rBMSCs were isolated from 4 week –old rats and cultured to the third generation by the adhesive method. According to the experimental purposes, the cells were inoculated in four groups: control, Y inhibitors-added, Sr-added Y inhibitors plus Sr-added . After cultured for 1, 4, and 7days respectively, cell proliferation was determined by CCK-8 kits, alkaline phosphatase activity was measured by use ALP kits Specific transcription factor group of cells Runx-2 and downstream bone morphogenic protein OCN,OPN etc to promote osteogenesis differentiation of gene expression . Results: First, cell prolifer -ation was promoted Rho kinase inhibitor Y-27632 compared to the blank group; Rho kinase inhibitor Y-27632 in Sr to rBMSCs, to which cell growth situation is faster than adding Sr group inpidually. Second, compared to the blank group, the ALP activity of rBMSCs increased modestly in Rho kinase inhibitor Y-27632-added group; In Rho kina -se inhibitor and Sr-dded group, the ALP activity of rBMSCs increased significantly fas -ter than that in Sr-added group. Third, compared to the blank group, the quantity of ge -ne expression significantly increased in Rho kinase inhibitor Y-27632-added group; the expression quantity of bone relating gene of rBMSCs had an especially significant incre -ases in Rho kinase inhibitor and Sr-dded group.Conclusion: Rho kinase inhibitor Y-27 -632 play an important role in osteogenic differentiation of rat bone marrow mesenchy -mal stem cells induced by strontium ranelate.     

 Key words : Bone marrow mesenchymal stem cells  Rho kinase inhibitor

Strontium ranelate  Proliferation  Gene expression

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引言 1