益坤饮通过ERα-SDF-1/CXCR4轴活化内皮祖细胞修复受损的血管内皮细胞OA
Yikunyin Activated Endothelial Progenitor Cells via ERα-SDF-1/CXCR4 Axis to Repair Injured Vascular Endothelial Cells
目的:探讨益坤饮通过ERα-SDF-1/CXCR4 轴介导内皮祖细胞(EPCs)的增殖、迁移及归巢,修复受损血管内皮细胞的作用机制.方法:培养鉴定大鼠骨髓来源的EPCs,建立肿瘤坏死因子α(TNF-α)诱导致内皮祖细胞(EPCs)损伤模型,加入不同浓度的益坤饮含药血清,分别加入雌激素受体α(ERα)抑制剂MPP、SDF-1/CX-CR4 抑制剂AMD3100 干预,MTT法测定细胞增殖;Transwell法检测细胞迁移;ELISA法测定细胞NO、VEGF、Ang-1、Ang-2 和SDF-1 含量;Matrigel管形成法测定EPCs管形成能力;Western blot法分别测定ERα、SDF-1、CXCR4 蛋白表达.结果:与空白对照组相比,模型对照组EPCs增殖率、迁移率降低(P<0.01),NO、VEGF、Ang-1、Ang-2 和SDF-1 含量降低(P<0.01),血管形成能力降低(P<0.01),ERα、基质细胞衍生因子(SDF-1)、C-X-C趋化因子受体4(CXCR4)蛋白表达明显下调;与模型对照组相比,13.6 g/kg益坤饮5%、10%、20%含药血清组EPCs的增殖率、迁移率升高,NO、VEGF、Ang-1、Ang-2 和SDF-1 分泌增加(P<0.05 或P<0.01),管总长和分支数增加(P<0.05或P<0.01),ERα、SDF-1、CXCR4 表达显著上调(P<0.01).加入抑制剂AMD3100 和MPP干预后,与空白对照组相比,MPP组和AMD3100 组EPCs增殖、迁移数目降低、NO、VEGF、Ang-1、Ang-2 和SDF-1 含量降低,管形成能力降低(P<0.05 或P<0.01),ERα、SDF-1、CXCR4 蛋白表达下调(P<0.01);与13.6 g/kg益坤饮20%含药血清组相比,MPP+益坤饮20%含药血清组及AMD3100+益坤饮 20%含药血清组的增殖、迁移率降低,分泌的NO、VEGF、Ang-1、Ang-2 和SDF-1 含量降低(P<0.05 或P<0.01),血管形成能力减弱(P<0.05),MPP+益坤饮组和AMD3100组SDF-1、CXCR4 表达下调(P<0.01),MPP+益坤饮组的Erα表达显著下调(P<0.01).结论:益坤饮通过调控ERα-SDF-1/CXCR4 轴介导EPCs的增殖、迁移及归巢,修复受损的内皮细胞.
Objective:To investigate the mechanism of Yikunyin(益坤饮,YKY)in mediating the proliferation,mi-gration,and homing of endothelial progenitor cells(EPCs)through the estrogen receptor α(ERα)-stromal cell-derived factor(SDF-1)/chemokine receptor 4(CXCR4)axis to repair injured vascular endothelial cells.Methods:Rat bone marrow-derived EPCs were cultured and identified.A tumor necrosis factor α(TNF-α)-induced EPC injury model was established.Serum containing YKY of different concentrations was added,along with ERα inhibitor MPP and SDF-1/CXCR4 inhibitor AMD3100.MTT was used to detect cell proliferation,and Transwell was used to detect cell migration.The levels of NO,VEGF,Ang-1,Ang-2,and SDF-1 were measured by ELISA,and the tube formation capacity was meas-ured by the Matrigel tube formation method,while protein expressions of ERα,SDF-1,and CXCR4 were detected by Western blot.Results:Compared with the blank control group,the model control group showed a significant decrease in the proliferation and migration of EPCs,as well as in the contents of NO,VEGF,Ang-1,Ang-2,and SDF-1,the angiogen-esis ability,and the protein expressions of ERα,SDF-1,and CXCR4.Compared with the model control group,the serum groups containing 13.6 g/kg YKY of different concentrations significantly promoted the proliferation and migration of EPCs,increased the secretion of NO,VEGF,Ang-1,Ang-2,and SDF-1,and significantly upregulated the expression levels of total tube length,branch number,ERα,SDF-1,and CXCR4(P<0.01).After intervention with the inhibitors AMD3100 and MPP,compared with those in the blank control group,the proliferation and migration numbers of EPCs,the contents of NO,VEGF,Ang-1,Ang-2,and SDF-1,as well as the tube formation ability and the protein expressions of ERα,SDF-1,and CXCR4,in the MPP group and the AMD3100 group all decreased(P<0.01).Compared with the 20%serum group containing 13.6 g/kg YKY,the intervention effects of MPP+20%serum group containing YKY and the AMD3100+20%serum group containing YKY showed a decrease in the proliferation and migration numbers,as well as the contents of NO,VEGF,Ang-1,Ang-2,and SDF-1 secreted.The ability to promote vascular formation was weakened.The expression of SDF-1 and CXCR4 was downregulated in the MPP+YKY group and the AMD3100 group(P<0.01),and the expression of Erα was significantly downregulated in the MPP+YKY group.Conclusion:YKY can repair in-jured endothelial cells by regulating the proliferation,migration,and homing of EPCs through the ERα-SDF-1/CXCR4 axis.
赵欣妍;李丽萍;詹群;陈霞;杜秋
南京中医药大学附属南京中医院,南京 210022南京中医药大学附属南京中医院,南京 210022南京中医药大学附属南京中医院,南京 210022南京中医药大学附属南京中医院,南京 210022南京中医药大学附属南京中医院,南京 210022
益坤饮内皮祖细胞雌激素受体α基质细胞衍生因子/C-X-C趋化因子受体4
YikunyinEndothelial progenitor cellERαSDF-1/CXCR4
《中药药理与临床》 2026 (1)
27-36,10
江苏省自然科学基金青年项目(编号:BK20210035)南京市中医药卫生青年人才项目(编号:ZYQ20012)南京市中药制剂与临床药学研究医学重点实验室建设项目(编号:NJSYXZDSYS-2023).
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