Objective： To investigate the effects of panaxadiol saponins component （PDS-C） isolated from total saponins of panax ginseng on proliferation, differentiation and corresponding gone expression profile of megakaryoc...Objective： To investigate the effects of panaxadiol saponins component （PDS-C） isolated from total saponins of panax ginseng on proliferation, differentiation and corresponding gone expression profile of megakaryocytes. Methods： Bone marrow culture of colony forming assay of megakaryocytic progenitor cells （CFU-MK） was observed for the promoting proliferation mediated by PDS-C, and differentiation of megakaryocytic blasts caused by PDS-C was analyzed with flow cytometry in CHRF-288 and Meg-01 cells, as well as proliferation, differentiation-related genes expression profile and protein expression levels were detected by human gone expression microarray and western blot. Results： In response to PDS-C 10, 20 and 50 mg/L, CFU-MK from 10 human bone marrow samples was increased by 28.9± 2.7%, 41.0% ± 3.2% and 40.5% ± 2.6% over untreated control, respectively （P〈0.01, each）. Flow cytometry analysis showed that PDS-C treated CHRF-288 cells and Meg-01 cells significantly increased in CD42b, CD41, TSP and CD36 positive ratio, respectively. PDS-C induced 29 genes up-regulated more than two-fold commonly in both cells detected by human expression microarray representing 4000 known genes. The protein expression levels of ZNF91, c-Fos, BTF3a, GATA-1, RGS2, NDRG2 and RUNX1 were increased with western blot in correspond to microarray results. Conclusion： PDS-C as an effective component for hematopoiesis, play the role to enhance proliferation and differentiation of megakaryocytes, also up-regulated expression of proliferation, differentiation-related genes and proteins in vitro. KEYWORDS panaxadiol saponins, megakaryocyte, gone expression profile, proliferation, differentiation展开更多
目的研究丹参酮ⅡA对血小板及骨髓巨核细胞的影响,探讨其抗血小板作用与其对骨髓巨核细胞影响的关系,了解丹参酮ⅡA是否对免疫性血管炎血管损害有保护作用。方法建立免疫性血管炎模型,实验分为正常对照组、模型对照组和丹参酮ⅡA治疗组...目的研究丹参酮ⅡA对血小板及骨髓巨核细胞的影响,探讨其抗血小板作用与其对骨髓巨核细胞影响的关系,了解丹参酮ⅡA是否对免疫性血管炎血管损害有保护作用。方法建立免疫性血管炎模型,实验分为正常对照组、模型对照组和丹参酮ⅡA治疗组,检测外周血血小板数量,同时检测巨核细胞的数量和功能(CFU-MK集落形成能力)以及基质细胞集落(CFU-F)形成能力,HE染色、弹力纤维染色及电镜观察血管病理改变。结果免疫性血管炎模型对照组有血小板数量、骨髓巨核细胞数、CFU-MK数的明显增加。丹参酮ⅡA治疗能够明显抑制血小板,血小板数量由模型组的931.33±254.19×109/L降至544.00±83.88×109/L(P<0.05)。也能明显抑制骨髓巨核细胞数和CFU-MK,分别为29.17±8.40个/片vs 17.67±5.75个/片(P<0.05)和33.0±14.27个/2×105cells vs 14.67±3.67个/2×105cells(P<0.05)。还能抑制骨髓基质细胞,CFU-F数由58.7±14.5个/2×106cells vs 38.8±11.09个/2×106cells(P<0.05)。丹参酮ⅡA治疗后血管炎的病理损害有明显减轻。结论丹参酮ⅡA可抑制血小板和骨髓巨核细胞的数量,对骨髓巨核细胞和基质细胞的抑制可能是丹参酮ⅡA抑制血小板功能的作用机理之一。丹参酮ⅡA可能通过抑制血小板和巨核细胞而减轻炎症损害,从而使免疫性血管炎血管免受损害而达到治疗目的。展开更多
基金Supported by National Natural Science Foundation of China(No.81373876)Zhejiang Provincial Natural Science Foundation(No.LY14H290004)Science and Technology Program of Zhejiang Province(No.2010C33098)
文摘Objective： To investigate the effects of panaxadiol saponins component （PDS-C） isolated from total saponins of panax ginseng on proliferation, differentiation and corresponding gone expression profile of megakaryocytes. Methods： Bone marrow culture of colony forming assay of megakaryocytic progenitor cells （CFU-MK） was observed for the promoting proliferation mediated by PDS-C, and differentiation of megakaryocytic blasts caused by PDS-C was analyzed with flow cytometry in CHRF-288 and Meg-01 cells, as well as proliferation, differentiation-related genes expression profile and protein expression levels were detected by human gone expression microarray and western blot. Results： In response to PDS-C 10, 20 and 50 mg/L, CFU-MK from 10 human bone marrow samples was increased by 28.9± 2.7%, 41.0% ± 3.2% and 40.5% ± 2.6% over untreated control, respectively （P〈0.01, each）. Flow cytometry analysis showed that PDS-C treated CHRF-288 cells and Meg-01 cells significantly increased in CD42b, CD41, TSP and CD36 positive ratio, respectively. PDS-C induced 29 genes up-regulated more than two-fold commonly in both cells detected by human expression microarray representing 4000 known genes. The protein expression levels of ZNF91, c-Fos, BTF3a, GATA-1, RGS2, NDRG2 and RUNX1 were increased with western blot in correspond to microarray results. Conclusion： PDS-C as an effective component for hematopoiesis, play the role to enhance proliferation and differentiation of megakaryocytes, also up-regulated expression of proliferation, differentiation-related genes and proteins in vitro. KEYWORDS panaxadiol saponins, megakaryocyte, gone expression profile, proliferation, differentiation
文摘目的研究丹参酮ⅡA对血小板及骨髓巨核细胞的影响,探讨其抗血小板作用与其对骨髓巨核细胞影响的关系,了解丹参酮ⅡA是否对免疫性血管炎血管损害有保护作用。方法建立免疫性血管炎模型,实验分为正常对照组、模型对照组和丹参酮ⅡA治疗组,检测外周血血小板数量,同时检测巨核细胞的数量和功能(CFU-MK集落形成能力)以及基质细胞集落(CFU-F)形成能力,HE染色、弹力纤维染色及电镜观察血管病理改变。结果免疫性血管炎模型对照组有血小板数量、骨髓巨核细胞数、CFU-MK数的明显增加。丹参酮ⅡA治疗能够明显抑制血小板,血小板数量由模型组的931.33±254.19×109/L降至544.00±83.88×109/L(P<0.05)。也能明显抑制骨髓巨核细胞数和CFU-MK,分别为29.17±8.40个/片vs 17.67±5.75个/片(P<0.05)和33.0±14.27个/2×105cells vs 14.67±3.67个/2×105cells(P<0.05)。还能抑制骨髓基质细胞,CFU-F数由58.7±14.5个/2×106cells vs 38.8±11.09个/2×106cells(P<0.05)。丹参酮ⅡA治疗后血管炎的病理损害有明显减轻。结论丹参酮ⅡA可抑制血小板和骨髓巨核细胞的数量,对骨髓巨核细胞和基质细胞的抑制可能是丹参酮ⅡA抑制血小板功能的作用机理之一。丹参酮ⅡA可能通过抑制血小板和巨核细胞而减轻炎症损害,从而使免疫性血管炎血管免受损害而达到治疗目的。
