摘要
目的探讨鹿茸通过磷脂酰肌醇3-激酶(PI3K)/蛋白激酶B(Akt)信号通路对内皮祖细胞(EPCs)活性及其细胞迁移的作用机制。方法将大鼠分为8组:空白组、模型组、鹿茸低剂量组、鹿茸中剂量组、鹿茸高剂量组、LY294002+鹿茸低剂量组、LY294002+鹿茸中剂量组、LY294002+鹿茸高剂量组。采用动脉分支分离法+维生素D3注射联合高脂喂养法建立下肢动脉硬化闭塞大鼠模型。鹿茸低剂量组、中剂量组、高剂量组分别给予相应剂量鹿茸灌胃给药,空白组、模型组给予等容量蒸馏水灌胃,LY294002+鹿茸低剂量组、LY294002+鹿茸中剂量组、LY294002+鹿茸高剂量组先行腹腔注射LY2940021 mg/(kg·d),再给予相应剂量鹿茸灌胃,连续干预28 d。流式细胞术检测外周血和骨髓中EPCs的CD133阳性细胞(CD133+)比例;酶联免疫吸附实验(ELISA)检测大鼠外周血EPCs中血管内皮生长因子(VEGF)、基质细胞衍生因子-1α(SDF-1α)水平;免疫荧光法检测大鼠腓肠肌中内皮型一氧化氮合酶(eNOS)、趋化因子受体4(CXCR4)阳性细胞的表达比例;蛋白质印迹(Western Blotting)法检测PI3K、Akt蛋白表达水平。结果与空白组比较,模型组及给药组骨髓与外周血中CD133+细胞比例升高;与模型组比较,各给药组CD133+细胞比例均降低,其中,鹿茸高剂量组高于LY294002+鹿茸高剂量组,LY294002+鹿茸高剂量组高于鹿茸中剂量组,鹿茸中剂量组高于LY294002+鹿茸中剂量组,LY294002+鹿茸中剂量组高于鹿茸低剂量组,鹿茸低剂量组高于LY294002+鹿茸低剂量组,差异均有统计学意义(P<0.05)。模型组及给药组VEGF、SDF-1α水平及eNOS、CXCR4阳性表达比例均较空白组降低,各给药组VEGF、SDF-1α水平及eNOS、CXCR4阳性表达比例均高于模型组,鹿茸高剂量组高于LY294002+鹿茸高剂量组,LY294002+鹿茸高剂量组高于鹿茸中剂量组,鹿茸中剂量组高于LY294002+鹿茸中剂量组,LY294002+鹿茸中剂量组高于鹿茸低剂量组
Objective To explore the mechanism of pilose antler on endothelial progenitor cell activity and cell migration through phosphatidylinositol 3-kinases(PI3K)/protein kinase B(Akt)signal pathway.Methods Rats were divided into 8 groups:normal group,model group,low dose pilose antler group,middle dose pilose antler group,high dose dose pilose antler group,LY294002+low dose pilose antler group,LY294002+middle dose pilose antler group,and LY294002+high dose pilose antler group.The rat model of arteriosclerosis obliterans of lower extremities was established by arterial branch separation and vitamin D3 injection combined with high-fat feeding.The rats in low,middle,and high dose groups of pilose antler were given corresponding doses of pilose antler by intragastric administration respectively.The rats in normal group and model group were given distilled water of the same volume.The rats in LY294002+low dose pilose antler group,LY294002+middle dose pilose antler group,and LY294002+high dose pilose antler group were first intraperitoneally injected with LY294002 and then intragastric administered with pilose antler for 28 days.The number of CD133+of endothelial progenitor cells(EPCs)in peripheral blood and bone marrow was detected by flow cytometry.The contents of stromal derived factor 1 alpha(SDF-1α)and vascular endothelial growth factor(VEGF)in serum and EPCs were detected by enzyme linked immunosorbent assay(ELISA).The expression proportion of endothelial nitric oxide synthase(eNOS)and chemokine receptor 4(CXCR4)positive cells in gastrocnemius muscle was detected by immunofluorescence.The expression levels of PI3K and Akt were detected by Western Blotting.Results The proportion of CD133+cells in model group and the administration group was higher than that in normal group.Compared with model group,the proportion of CD133+cells in administration groups was decreased,and the proportion of CD133+cells in the administration group was significantly higher than that in the corresponding administration+inhibitor groups(P<0.0
作者
韩杰
李廷荃
李子娟
王雁彬
霍凯
HAN Jie;LI Tingquan;LI Zijuan;WANG Yanbin;HUO Kai(Shanxi University of Traditional Chinese Medicine,Taiyuan 030024,Shanxi,China)
出处
《中西医结合心脑血管病杂志》
2022年第6期997-1002,1007,共7页
Chinese Journal of Integrative Medicine on Cardio-Cerebrovascular Disease
基金
国家自然科学基金项目(No.81874474)。
关键词
下肢动脉硬化闭塞症
鹿茸
磷脂酰肌醇3-激酶
蛋白激酶B
内皮祖细胞
实验研究
arteriosclerosis obliterans of lower extremities
pilose antler
phosphatidylinositol 3-kinases
protein kinase B
endothelial progenitor cells
experiment research
