目的:了解黄芪注射液对内皮细胞增殖及分泌黏附分子 VCAM-1 和 E-selectin 的影响,探讨黄芪注 射液对造血调控的机理。 方法: (1). 建立人脐静脉内皮细胞体外培养模型,用免疫细胞化学方法及电子显微镜 技术鉴定内皮细胞;(2). 将 HUVEC 与黄芪注射液不同剂量(0滋g / mL、4滋g / mL、40滋g / mL、400滋g / mL 和 4000滋g / mL)一起培养,用 MTT 法检测内皮细胞增殖;以流式细胞分析术测定内皮细胞增殖周期;以 TNF 作为阳性对照, 用 ELISA 法对黏附分子 VCAM-1 和 E-selectin 进行测定。 结果: (1)成功的建立了人脐静脉内皮细胞的体外模 型;用峪因子相关抗原免疫细胞化学染色结果为阳性、电镜检测到 W-P 小体,证实培养细胞为人脐静脉内皮细 胞。 (2)内皮细胞随着培养时间的延长,各组均显示出不同程度的促生长效应(Time:P<0. 05;Concentration:P< 0. 05)。 (3)细胞周期检测显示实验组内皮细胞周期各时相的细胞数占细胞总数的百分比与黄芪组比较有明显 的不同(P<0. 05)。 S 期和 G 2 / M 期细胞显著增多(P<0. 05),而 G 0 / G 1 期细胞相对比例减少(P<0. 05)。 (4) 体外培养的内皮细胞可自然分泌 VCAM-1。 各个浓度的黄芪注射液与 TNF 一样均具有促进内皮细胞分泌 VCAM-1 的作用,其中400滋g / mL 的黄芪注射液浓度是促进内皮细胞分泌 VCAM-1 的最佳浓度(P<0. 05),而且 黄芪注射液具有协同 TNF 的作用(P<0. 05)。 (5)内皮细胞体外培养在没有任何刺激的情况下未检测到 E- selectin. 黄芪注射液刺激内皮细胞也未检测到 E-selectin. 内皮细胞在 TNF 刺激活化后表达 E-selectin. 黄芪注射 液能促进 TNF 活化的内皮细胞分泌 E-selectin(P<0. 05)。 结论: 本研究提示,黄芪的补气生血途径之一可能是 通过直接刺激内皮细胞增殖和分泌造血生长因子和黏附分子,从而间接发挥造血调控作用的;黄芪可能促进造 血干祖细胞的归巢,并为开发黄芪用于辅助治疗血液病及肿瘤放、化疗后骨髓重建提供了理论依据。

Objective:To investigate the effect of Huangqi injection on proliferation and expression of vascular cell adhesion molecule and E - selectin in human umbilical vein endothelial cells (HUVEC). To study the mechanism of Huangqi injiection on regulation of hematopoiesis. Methods: (1)Human umbilical vein endothelial cells culture model was founded,Endothelial cells was identified by immunohistochemical staining and transmission electron microscope ( 2 ) Treating HUVEC withvarious concentration of Huangqi injiection(0滋g / mL,4滋g / mL,40滋g / mL,400滋g / mL,4000滋 g / mL). Analyzed the proliferation of HUVEC by MTT;Analyzed the proliferation phase of HUVEC using flow cytometry,and measure the expression of VCAM and E-selectin by ELISA. Results:(1)The results of immunohistochemical staining on correlation antigens of VIII factor and Weible-Paladabody in the en鄄 dothelial cells was observed by using transmission electron microscope. demonstrat that we established the human umbilical vein endothelial cells culture model. (2)Every groups has different influences on cell growth( Time:P = 0. 000 <0. 05;Concentration:P<0. 05). (3) There was significant change was found between the experiment group and the control group on the proliferation phase of HUVEC(P< 0. 05). The number of cells in S phase and G 2 / M phase were significant increased,(P<0. 05). The number of cells in G 0 / G 1 phase were reduced. (4)Endothelial cells were cultured in vitro can secret VCAM-1 no stimulation. Different Huangqi injection and TNF can promoted the secretion of VCAM-1 from endothelial cells. 400滋g / mL Huangqi injection is the most proper concentration. ( P < 0. 05), meanwhile,Huangqi injection can promoted the roles of TNF(P <0. 05). (5) Endothelial cells were cultured in vitro can not secret E - selectin no stimulation. Huangqi injection can not promoted the secretion of E-selectin from endothelial cells directly. Endothelial cells can been activated by TNF, Huangqi injection can promoted the secretion of E-selectin from activated endothelial cells(P<0. 05). Conclusions: This research suggest that Huangqi injection can support hematopoiesis through endothelial cells and adhesion molecule,Huangqi injection may promote the homing of hematopoietic stem/ progenitor cells. Theory basis was afforded for the clinical application of Huangqi injection.

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