祛瘀生肌方含药血清对人真皮微血管内皮细胞迁移和PGT相关因子的影响

Effects of Quyu Shengji Formula Containing Serum on the Migration of Human Dermal Microvascular Endothelial Cells and Expressions of PGT-related Factors

目的探讨祛瘀生肌方对人真皮微血管内皮细胞(HDMECs)迁移能力影响和前列腺素转运蛋白(PGT)相关因子的作用机制。方法将HDMECs细胞分为5组。低糖空白组(A组):细胞中加5mmol/L葡萄糖+10%空白血清;高糖空白组(B组):细胞中加25mmol/L葡萄糖+10%空白血清;生肌药组(C组):细胞中加25mmol/L葡萄糖+10%生肌药含药血清;化瘀药组(D组):细胞中加25mmol/L葡萄糖+10%化瘀药含药血清;生肌化瘀药组(E组):细胞中加25mmol/L葡萄糖+10%生肌化瘀药含药血清。采用Transwell、划痕实验、ELISA、Western Blot检测含药血清干预后HDMECs的迁移能力和PGT相关因子的表达。结果Transwell结果示:与B组比较,C、D、E组的细胞迁移能力明显升高(P<0.05)。划痕实验0、12、24 h显示,D组未愈合面积进一步缩小(P<0.05)。ELISA结果示C、D、E组的前列腺素E2(PGE2)含量均有所增加,其中D组的PGE2含量最接近低A组。Western Blot结果示:与B组比较,给药后C、D、E组PGT含量均显著降低(P<0.05),细胞内皮生长因子(VEGF)分泌量均显著增加(P<0.05),D组增加量最为显著(P<0.01)。结论祛瘀生肌方含药血清可提高HDMECs迁移能力,促进HDMECs分泌PGE2、VEGF,并降低PGT含量。

Objective To observe the effects and the mechanism of Quyu Shengji Formula(QYSJF)on the migration of human dermal microvascular endothelial cells(HDMECs)and expressions of prostaglandin transporter(PGT)-related factors.Methods HDMECs were divided into 5 groups:low glucose blank group(A)[b y adding 5 mmol/L glucose+10%vehicle serum],high glucose blank group(B)[b y adding 25 mmol/L glucose+10%vehicle serum],Shengji Formula group(C)[by adding 25 mmol/L glucose+10%Shengji Formula containing serum],Quyu Formula group(D)[by adding 25 mmol/L glucose+10%Quyu Formula containing se ru m],QYSJF group[b y adding 25 mmol/L glucose+10%QYSJF containing se ru m].Effects of serum containing QYSJF on the migration ability of HDMECs and the expressions of PGT-related factors were detected by Transwell experiment,scratch test,ELISA,and Western Blot.Results Transwell results showed that compared with Group B,the cell migration ability significantly increased in Group C,D,and E(P<0.05).The scratch test at 0,12,and 24 h showed that the unhealed area in Group D was further reduced(P<0.05).ELISA results showed that the content of PGE2 increased in Group C,D,and E.Of them,PGE2 content in Group D was the closest to Group A.Western Blot results showed that the PGT content was significantly lower after administration in Group C,D,and E,as compared with Group B(P<0.05).Compared with Group B,the secretion of vascular endothelial growth factor(VEGF)increased significantly after administration in Group C,D,and E(P<0.05),and the increase was the most significant in Group D(P<0.01).Conclusion QYSJF containing serum improved the migration ability of HDMECs,promoted its secretions of PGE2 and VEGF,and decreased the content of PGT.