Apelin-13在改善自体动静脉内瘘血管狭窄中的机制

目的探讨内源性配体13(Apelin-13)在改善自体动静脉内瘘血管狭窄中的机制。方法将该院收治的30例自体动静脉内瘘血管狭窄患者纳入观察组,30例自体动静脉内瘘非血管狭窄患者纳入对照组,检测所有研究对象血清中Apelin-13、血管紧张素Ⅱ(AngⅡ)、一氧化氮(NO)水平。在细胞水平,采用Apelin-13干扰序列(shApelin组)和无义序列(Scramb组)转染人脐静脉内皮细胞(HUVECS);采用AngⅡ、Apelin-13刺激HUVECS,并通过免疫印迹试验检测细胞中磷酸化内皮型一氧化氮合酶(peNOS)和非磷酸化内皮型一氧化氮合酶(eNOS)的表达情况。同时检测细胞培养上清液中NO水平。结果观察组血清Apelin-13水平明显低于对照组,差异有统计学意义(t=13.23,P<0.05);观察组血清中AngⅡ水平高于对照组,差异有统计学意义(t=10.45,P<0.05)。观察组NO水平低于对照组,差异有统计学意义(t=9.83,P<0.05)。shApelin组的Apelin-13、peNOS表达量低于Scramb组(t=19.82、83.28,P<0.05),但两组eNOS表达量差异无统计学意义(t=1.52,P>0.05)。shApelin组NO水平明显高于Scramb组,差异有统计学意义(t=10.37,P<0.05)。免疫印迹试验结果显示,与未进行AngⅡ刺激的HUVECS对比,AngⅡ刺激后HUVECS的peNOS表达量降低(t=1.02,P<0.05),而eNOS表达量差异无统计学意义(t=0.60,P>0.05)。未进行AngⅡ刺激的HUVECS与AngⅡ刺激后的HUVECS细胞中NO的水平比较,差异有统计学意义(t=5.58,P<0.05);AngⅡ刺激后的HUVECS中peNOS表达量与AngⅡ、Apelin-13同时刺激的HUVECS中peNOS表达量比较,差异有统计学意义(t=16.04,P<0.05),但eNOS表达量差异无统计学意义(t=1.71,P>0.05)。AngⅡ及Apelin-13同时刺激所培养细胞的上清液中NO水平低于AngⅡ刺激的NO水平(t=5.97,P<0.05)。结论 Apelin-13、AngⅡ通过共同刺激和介导peNOS/NO的信号通路发挥调控作用,从而改善血管狭窄情况。

Mechanism of Apelin-13 in improvement of autogenous arteriovenous fistula patients with angiostenosis

Objective To discuss the mechanism of endogenous ligand 13(Apelin-13)in improvement of autogenous arteriovenous fistula patients with angiostenosis.Methods A total of 30 autogenous arteriovenous fistula patients with angiostenosis were selected as the observation group and 30 autogenous arteriovenous fistula patients without angiostenosis were selected as the control group.The levels of Apelin-13,angiotensinⅡ(AngⅡ),nitric oxide(NO)were detected.At the cellular level,interference sequence of Apelin-13(shApelin group)and nonsense sequence(Scramb group)were used to transfect human umbilical vein endothelial cells(HUVECS).At the same time,AngⅡand Apelin-13 were used to stimulate the HUVECS,in addition,phosphorylated endothelial nitric oxide synthase(peNOS)and unphosphorylated endothelial nitric oxide synthase(eNOS)were detected by Western blot.In the supernatant of cell culture,the levels of NO were detected.Results The serum Apelin-13 level in the observation group was significantly lower than that of the control group(t=13.23,P<0.05).The level of AngⅡin the observation group was significantly higher than that of the control group(t=10.45,P<0.05).The NO level of the observation group was significantly lower than that of the control group(t=9.83,P<0.05).Apelin-13 and peNOS expression levels of shApelin group were significantly lower than that of Scramb group(t=19.82 and 83.28,P<0.05),but the difference in eNOS expression between the two groups was not statistically significant(t=1.52,P>0.05).The NO level in the shApelin group was significantly higher than Scramb group(t=10.37,P<0.05).Western blot showed that compared with HUVECS without AngⅡstimulation,peNOS expression level of HUVECS had decreased after AngⅡstimulation(t=0.60,P<0.05),while eNOS expression level had no statistically significant difference(t=1.02,P>0.05).The levels of NO produced by HUVECS without AngⅡstimulation had statistically significant difference with HUVECS stimulated by AngⅡ(t=5.58,P<0.05).PeNOS expression produced by HUVECS after stimulation by AngⅡhad statistically significant difference with HUVECS cells stimulated by AngⅡand Apelin-13 at the same time(t=16.04,P<0.05),but the difference of eNOS expression was not statistically significant(t=1.71,P>0.05).The NO level in the supernatant cultured by AngⅡand Apelin-13 at the same time was significantly lower than that of only stimulated by AngⅡ(t=5.97,P<0.05).Conclusion By stimulating the peNOS/NO signal pathway together,Apelin-13 and AngⅡcould effectively improve the vascular stenosis.