cAMP/Ca^(2+)信号通路在红景天苷诱导骨髓间充质干细胞向神经元细胞定向分化中的作用和机制

Role and mechanism of cAMP/Ca^(2+) signal pathway in differentiation of bone marrow mesenchymal stem cells into neuronal cells induced by salidroside

本研究旨在探讨cAMP/Ca^(2+)信号通路在红景天苷诱导骨髓间充质干细胞(BMSCs)向神经元细胞定向分化中的作用机制。将骨髓间充质干细胞株D1细胞分为空白对照组和红景天苷诱导组,空白对照组细胞加入不含药物的等量D/F12完全培养液,诱导组细胞加入含有100 mg·L^(–1)红景天苷的D/F12完全培养液分别作用24、48和72 h。运用RT^2 Profiler PCR Array方法检测红景天苷诱导BMSCs不同时间后c AMP/Ca^(2+)信号通路中差异表达的基因。实验表明,红景天苷分别诱导BMSCs 24、48和72 h后,与空白对照组比较,表达有差异的基因共38个,其中上调基因23个,下调基因15个,经过筛选上调较显著的基因有4个:DNA损伤诱导因子3(Ddit3)、热休克蛋白5(Hspa5)、磷酸酶1调节亚单位15a(Ppp1r15a)、前列腺素内过氧化物合酶(Ptgs2);下调显著的基因有4个:胰高血糖素(Gcg)、白细胞介素2(Il2)、肿瘤坏死因子(Tnf)、生长激素抑制素(Sst)。这些基因可能对红景天苷诱导BMSCs向神经元细胞的定向分化具有一定影响。

To investigate the mechanism of cAMP/Ca^2＋ signaling pathway inducing bone marrow mesenchymal stem cells to differentiate into neuronal ceils, we cultured the bone marrow mesenchymal stem cells D1 cells in the present study. D 1 ceils were divided into two groups： control group and salidroside inducing groups. Control group was cultured with complete culture solution D/F12, while salidroside inducing groups were induced with 100 mg·L^-1 salidroside for different time periods （24, 48 and 72 hours）. PCR-array assay was used to detect expression of 84 calcium related mRNA, and significantly different genes were chosen to analyse. As a result, there were 4 significantly upregulated mRNAs including DNA damage-inducible transcript 3 （Ddit3）, heat shock protein 5 （Hspa5）, protein phosphatase 1 regulatory subunit （Ppp1r15a） and prostaglandin-endoperoxide synthase 2 （Ptgs-2）, and 4 significantly downregulated mRNAs including glucagon （Gcg）, interleukin 2 （I12）, tumor necrosis factor （Tnf） and somatostatin （Sst） in the cAMP/Ca^2＋ signaling pathway. They probably had an effect on the process of salidroside induced D1 cells differentiating into neuronal cells.