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目地:观察骨形态发生蛋白4(bone morphogenetic proteins-4,BMP4)对龟甲提取物(Extracts from Testudinis Carapacis et Plastri,PTE)促骨髓间充质干细胞(mesenchymal stem cells,MSCs)增殖的影响及作用机制。方法:PGL3-ID1分别和0、0.1、0.3、0.5、1μg/m L的p EGFP-BMP4用磷酸钙共沉淀法共转染大鼠MSCs,转染细胞每个剂量组均分为空白对照组和PTE组。空白对照组不加PTE,PTE组加入30μg/m L的PTE作用36 h,收集细胞萤光素酶报告基因检测ID1的表达。选取0.3μg/m L的p EGFP-BMP4与PGL3-ID1共转染。再将MSCs细胞分为对照组、PTE组、BMP4组和BMP4+PTE组。对照组和PTE组MSCs不做转染,PTE组MSCs加入30μg/m L的PTE作用36h;BMP4组MSCs共转染PGL3-ID1和p EGFP-BMP4;BMP4+PTE组MSCs共转染PGL3-ID1和p EGFP-BMP4,加入30μg/m L的PTE作用36 h。应用RT-PCR分别检测各组中ID1、BMP4、RARα的表达。结果:与空白对照组比较PTE组BMP4、ID1、RARα表达均显著增高;BMP4组BMP4、RARα表达显著增高,ID1表达显著降低;BMP4+PTE组BMP4、ID1、RARα表达与BMP4组相比均显著降低。结论:龟甲提取物可以促进MSCs增殖,其又可通过调节BMP4的表达防止MSCs的过度增殖。
Objective: To investigate the effects of bone morphogenetic proteins-4 (BMP4) on the proliferation of mesenchymal stem cells (MSCs) induced by Extracts from Testudinis Carapacis et Plastri (PTE) and its mechanism of action . METHODS: PGL3-ID1 was co-transfected into 0, 0.1, 0.3, 0.5 and 1 μg / mL of p EGFP-BMP4 by calcium phosphate co-precipitation method. The transfected cells were divided into blank control group And PTE group. Blank control group without PTE, PTE group 30μg / mL of PTE for 36 h, collected cells luciferase reporter gene detection ID1 expression. Select 0.3μg / m L of p EGFP-BMP4 co-transfected with PGL3-ID1. MSCs were divided into control group, PTE group, BMP4 group and BMP4 + PTE group. PGCs were co-transfected with PGL3-ID1 and p EGFP-BMP4 in MSCs of BMP4 group and MSCs of BMP4 + PTE group co-transfected with PGL3-ID1 And p EGFP-BMP4, adding 30μg / mL of PTE for 36h. The expression of ID1, BMP4 and RARα in each group were detected by RT-PCR. Results: Compared with the blank control group, the expression of BMP4, ID1 and RARα in PTE group were significantly increased; the expression of BMP4 and RARα in BMP4 group was significantly increased and the expression of ID1 was significantly decreased; BMP4, ID1 and RARα expression in BMP4 + PTE group were significantly higher than those in BMP4 group reduce. Conclusion: Tortoise shell extract can promote the proliferation of MSCs, which in turn can prevent the excessive proliferation of MSCs by regulating the expression of BMP4.