HDPSCs在体外促进血管再生的潜能及其分子机理研究

Study on the potential of HDPSCs in promoting angiogenesis in vitro and its molecular mechanism

目的探讨人牙髓干细胞(HDPSCs)在体外促进血管再生的潜能及其机制。方法体外分离培养HDPSCs,同时选取人微血管内皮细胞(HMEC)随机分为阴性对照组,阳性对照组和HDPSCs组,其中阴性对照组采用不含胎牛血清(FBS)的-MEM培养基,阳性对照组加入含100ml/l FBS的-MEM培养基,HDPSCs组加入HDPSCs培养。采用MTS法检测HEMC细胞增殖情况,采用Trans-well小室实验检测HEMC细胞迁移能力,采用Western blot法检测ERK1/2、p-ERK1/2、p38和p-p38蛋白表达,并对比各组检测结果。结果阳性对照组培养24h和48h HEMC细胞OD值分别为(0.511±0.099)和(0.624±0.103),明显高于阴性对照组和HDPSCs组(P<0.05);HDPSCs组培养24h和48h HEMC细胞OD值分别为(0.390±0.098)和(0.455±0.096),明显高于阴性对照组(P<0.05);阳性对照组HEMC细胞迁移数为(115.64±12.06)个,明显多于阴性对照组和HDPSCs组(P<0.05);HDPSCs组HEMC细胞迁移数为(75.60±9.81)个,明显多于阴性对照组(P<0.05);阳性对照组ERK1/2、p-ERK1/2、p38和p-p38蛋白相对表达量分别为(0.947±0.037)、(0.867±0.043)、(0.833±0.069)和(0.792±0.086),明显高于阴性对照组和HDPSCs组(P<0.05);HDPSCs组ERK1/2、pERK1/2、p38和p-p38蛋白相对表达量分别为(0.528±0.046)、(0.422±0.050)、(0.554±0.074)和(0.512±0.077),明显高于阴性对照组(P<0.05)。结论 HDPSCs在体外可促进血管内皮细胞增殖和迁移,可能与其参与ERK1/2和p-p38MAPK信号通路有关。

Objective To investigate the potential of human dental pulp stem cells（HDPSCs） promoting angiogenesis in vitro and its mechanism. Methods HDPSCs were isolated and cultured in vitro; and human microvascular endothelial cells（HMEC）were selected and randomly divided into negative control group, positive control group and HDPSCs group; the negative control group was added-MEM culture medium without fetal bovine serum（FBS）, the positive control group was added-MEM culture medium with 100 mL/L FBS, and HDPSCs group was added HDPSCs co-culture. MTS method was used to detect the proliferation of HEMC cells, the migration ability of HEMC cells was detected by scarification method, and the expression of p-ERK1/2 and p-p38 protein was detected by Western blot method. Results The OD values of HEMC cells cultured 24 h and 48 h in the positive control group were respectively（0.511±0.099）and（0.624±0.103）, which was significantly higher than that in the negative control group and HDPSCs group（P〈0.05）; The OD values of HEMC cells cultured 24 h and 48 h in the HDPSCs group were respectively（0.390±0.098） and（0.455 ±0.096）, which was significantly higher than that in the negative control group（P〈0.05）; The migration number of HEMC cells in positive control group was（115.64±12.06）, which was significantly higher than that in negative control group and HDPSCs group（P〈0.05）;The migration number of HEMC cells in HDPSCs group was（75.60±9.81）, which was significantly higher than that in negative control group（P〈0.05）;The relative expression of ERK1/2, p-ERK1/2, p38 and p-p38 protein in the positive control group were respectively（0.947 ±0.037）,（0.867 ±0.043）,（0.833±0.069） and（0.792±0.086）, which were significantly higher than those in the negative control group and HDPSCs group（P〈0.05）; The relative expression of ERK1/2, p-ERK1/2, p38 and p-p38 protein in the HDPSCs group were respectively（0.528 ±0.046）,（0.422 ±0.050）,（0.554 ±0.074） and（0.512 ±0.077）, which were significantly higher than those in the negative control group（P〈0.05）. Conclusion HDPSCs can promote the proliferation and migration of vascular endothelial cells in vitro, which may be related to its involvement of ERK1/2 and p-p38 MAPK signaling pathway.