PHLPP基因对血管损伤后狭窄的影响

Effect of PHLPP Gene on Stenosis after Vascular Injury

目的分析蛋白磷酸酶(PHLPP)基因表达水平对血管损伤后狭窄的影响,并探讨其可能机制。方法动物实验,大鼠分为研究组、对照组和假手术组。研究组大鼠鼠尾静脉给予PHLPP腺病毒,对照组大鼠鼠尾给予空载病毒。给予颈动脉球囊损伤,2周后取颈动脉,切片后染色,观察血管损伤后狭窄程度。细胞实验,取大鼠血管平滑肌原代细胞,分别将质粒pc DNA3.0/PHLPP(研究组)和pc DNA3.0/空载(对照组)转染至该细胞,采用EDU、划痕实验对细胞增殖和迁移能力进行检测;Western blot技术对增殖细胞核抗原(PCNA)蛋白、基质金属蛋白酶2(MMP2)蛋白和PHLPP基因进行检测。结果研究组的损伤血管后狭窄程度较对照组减轻,细胞增殖和迁移能力显著低于对照组,并且PCNA和MMP2的m RNA水平和蛋白水平低于对照组(P<0.05)。结论 PHLPP表达水平可通过调节血管平滑肌细胞增殖和迁移能力而影响损伤后血管的狭窄程度;抑制PCNA和MMP2表达可能为其作用机制。

Objective To explorer the effect of PHLPP gene express level on vasscular stenosis,and its mechanism.Methods Animal experiments,the rats were divided into study group,control group and sham operation group,rat tail vein of study group rats were given PHLPP adenovirus,rats of control group were given no-load virus,rats were given carotid artery balloon.The injured and sham-operated group only sutured the skin.The carotid artery was taken after 2 weeks and stained after slicing to observe the degree of stenosis after vascular injury.Primary cultured rat vascular smooth muscle cells were transfected with plasmid pcDNA3.0/PHLPP(study group)and pcDNA3.0/no-load(control group)respectively.The proliferating cell nuclear antigen(PCNA)protein,matrix metallopeptidase 2(MMP2)protein and PH domain leucine-rich repeat protein phosphatase(PHLPP)protein were detected by Western blot.Results The degree of vascular stenosis in study group was lessened compared with that of control group.The proliferation and migration of cells in study group were significantly lower than those in control group.The mRNA and protein levels of PCNA and MMP2 in study group were lower than those in control group(P<0.05).Conclusion The expression of PHLPP may affect the degree of vascular stenosis by regulating the proliferation and migration of vascular smooth muscle cells.Inhibiting the expression of PCNA and MMP2 may be the mechanism of action.