基于PI3K/Akt通路探讨鹿茸促进下肢动脉硬化闭塞症大鼠血管新生的作用机理

The Mechanism of Lurong Promoting Angiogenesis in Rats with Arteriosclerosis Obliterans of the Lower Extremity Based on PI3K/Akt Pathway

目的:基于磷脂酰肌醇3-羟基激酶(phosphatidylinositol 3-hydroxykinase,PI3K)/蛋白激酶B(protein kinase B,Akt)信号通路探讨鹿茸促进下肢动脉硬化闭塞症(arteriosclerosis obliterans of the lower extremities,ASOLE)大鼠血管新生的作用机理。方法:通过肢体缺血手术及高脂饲料喂养制作大鼠下肢动脉硬化闭塞症模型。应用酶联免疫吸附试验(Enzyme-linked immunosorbent assay,ELISA)方法检测大鼠血清中血管内皮生长因子(vascular endothelial growth factor,VEGF)、缺氧诱导因子1α多肽(hypoxia-inducible factor 1-alpha,HIF-1α)含量;TUNEL法检测大鼠骨髓中内皮祖细胞(endothelial progenitor cells,EPCs)凋亡率;免疫荧光方法检测大鼠腓肠肌中血管内皮细胞生长因子受体(vascular endothelial growth factor receptor,VEGFR)阳性细胞的表达比例。结果:血清中VEGF、HIF-1α含量及腓肠肌VEGFR阳性表达模型组及给药组低于空白组(P<0.05),给药组高于模型组(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,各组间比较差异有统计学意义(P<0.05);骨髓中EPCs凋亡率空白组及鹿茸高剂量组最低,两组比较差异无统计学意义(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EPCs凋亡率依次增高,组间比较差异有统计学意义(P<0.05)。结论:鹿茸可能通过PI3K/Akt信号通路促进EPCs增殖,进而促进ASOLE大鼠血管新生。

Objective:To explore the mechanism of Lurong(Cervi Cornu Pantotrichum)promoting angiogenesis in rats with arteriosclerosis obliterans of the lower extremities(ASOLE)based on PI3K/Akt signaling pathway.Methods:Rat models with ASOLE were prepared by limb ischemia surgery and high-fat diet.ELISA was applied to detect the contents of VEGF and HIF-1αin the serum;TUNEL method was used to measure the apoptosis rate of EPCs in bone marrow;immunofluorescence method to detect the expression of VEGFR positive cells in rat gastrocnemius muscles.Results:The contents of VEGF and HIF-1αin the serum and positive expressions of VEGFR in gastrocnemius muscles of the model group and medication group were lower than these of the blank group(P<0.05),the medication group was higher than the model group(P<0.05),among them,these increased in low dose of Lurong+LY294002 group,low dose group of Lurong,moderate dose of Lurong+LY294002 group,moderate dose group of Lurong,high dose of Lurong+LY294002 group,high dose group of Lurong,and the difference had statistical meaning between different groups(P<0.05);the apoptosis rates of EPCs in bone marrow were the lowest,and the difference had no statistical meaning(P>0.05),the apoptosis rate of EPCs in bone marrow increased successively in high dose of Lurong+LY294002 group,moderate dose group of Lurong,moderate dose of Lurong+LY294002 group,low dose group of Lurong,low dose of Lurong+LY294002 group and the model group,and the difference showed statistical meaning between different groups(P<0.05).Conclusion:Lurong could promote the proliferation of EPCs possibly through PI3K/Akt signaling pathway,and then promote angiogenesis of ASOLE rats.