鹿茸通过PI3K/AKT信号通路调控EPCs动员及归巢促进PAD大鼠血管新生的作用机理

Peripheral artery disease (PAD) in the lower extremity is a intractable disease that endangers the health of the elderly, which can cause gangrene at the extremities and lead to amputation.Exploring the mechanism of promoting angiogenesis in ischemic areas is of great significance to the treatment of PAD.Endothelial progenitor cells (EPCs) are derived from the bone marrow.EPCs can home to the ischemic regions and differentiated into mature endothelial cells, which are involved in angiogenesis.In the early stage of the study, velvet antler can promote the growth of angiogenesis, EPCs' proliferation and angiogenesis, but the specific mechanism is not clear.In this project,The experimental technique of RT-PCR,immunoblotting and immunofluorescence were used to analyze that how velvet regulate EPCs' mobilization and homing by signal transduction pathways and targeting mechanisms;the intrinsic material basis for the treatment of PAD is explained. In this study, the mechanism of antler-induced angiogenesis in PAD rats was deeply investigated, which provided a theoretical basis for its development and clinical application.

下肢动脉硬化性闭塞症（PAD）是危害老年人健康的难治性疾病，易引起肢体末端坏疽，导致截肢。探索促进机体缺血部位血管新生的机制对治疗PAD意义深远。内皮祖细胞（EPCs）来源于骨髓，具有缺血区定向归巢，分化为成熟内皮细胞进而参与血管新生的能力。鹿茸补肾、填精、益髓，前期研究提示，鹿茸能够促进PAD大鼠血管新生、EPCs增殖及促血管生成相关因子升高，但具体机制尚未明晰。本项目拟运用RT-PCR、免疫印迹、免疫荧光等实验技术，分析鹿茸参与调控EPCs动员及归巢的信号转导介入通路和靶向机制；同时阐释治疗PAD证治规律的内在物质基础。本研究深入探讨鹿茸促进PAD大鼠缺血下肢血管新生的作用机理，为其研发及临床运用提供理论依据。

下肢动脉硬化性闭塞症（PAD）是危害老年人健康的难治性疾病，易引起肢体末端坏疽，导致截肢。探索促进机体缺血部位血管新生的机制对治疗PAD意义深远。本项目主要研究运用流式细胞术、RT-PCR、免疫印迹及免疫荧光等技术及ELISA等技术检测鹿茸对PAD大鼠下肢EPCs 细胞增殖及PI3K/AKT信号通路的作用，测定PI3K/AKT信号通路及相关因子SDF-1α、eNOS、VEGF及MMP9等在鹿茸干预后的表达，进一步探索鹿茸调控EPCs动员、归巢及促进PAD血管新生的机制。结果证明PAD缺血区促血管新生作用与信号通路PI3K/AKT有着密切关系；鹿茸通过对PI3K/AKT信号通路及其关键因子、蛋白表达的干预，进而参与调控EPCs动员及归巢。实验结果发现CD133+细胞比例模型组及给药组均显著高于空白组（P<0.05），其中给药组均显著高于给药组+抑制剂组（P<0.05）；骨髓中EPCs凋亡率以空白组及鹿茸高剂量组最低，其二者无显著差异（P>0.05），鹿茸高剂量+LY294002组、鹿茸中剂量组、鹿茸中剂量+LY294002组、鹿茸低剂量组、鹿茸低剂量+LY294002组、模型组依次增高，之间均存在显著差异（P<0.05）。血清中VEGF、SDF-1α含量以及腓肠肌eNOS、CXCR4阳性表达中模型组及给药组显著低于空白组（P<0.05），给药组显著高于（P<0.05），其中鹿茸低剂量+LY294002组、鹿茸低剂量组、鹿茸中剂量+LY294002组、鹿茸中剂量组、鹿茸高剂量+LY294002组、鹿茸高剂量组依次增加，之间均存在显著差异（P<0.05）。本研究深入探讨鹿茸促进PAD大鼠缺血下肢血管新生的作用机理，为其研发及临床运用提供理论依据。