LncRNA p21调控Hippo-YAP信号通路对小鼠腹主动脉瘤形成的影响及机制

Effect of LncRNA p21 regulating Hippo-YAP signaling pathway on formation of abdominal aortic aneurysm in mice and its mechanism

目的 探究长链非编码RNA p21(long non-coding RNA,LncRNA p21)调控Hippo-Yes相关蛋白(Hippo-Yes-associated protein, Hippo-YAP)信号通路对小鼠腹主动脉瘤(abdominal aortic aneurysm, AAA)形成的影响。方法 C57BL/6 ApoE-/-通过皮下植入渗透性微泵构建血管紧张素Ⅱ(angiotensinⅡ,AngⅡ)诱导小鼠AAA模型。将C57BL/6 ApoE-/-随机分为假手术组(sham)、模型组(model)、LncRNA p21阴性对照组(sh-NC)、LncRNA p21敲低组(sh-LncRNA p21)。HE和血管弹力纤维染色(VVG)观察血管形态变化;qRT-PCR检测LncRNA p21表达;Western blot检测MMP-2、MMP-9、TIMP-1表达;原位末端标记法(TUNEL)染色检测平滑肌细胞凋亡;Western blot检测caspase-3、cyclinD1及Hippo-YAP信号通路蛋白表达。结果 与sham组相比,model组小鼠血管弹力纤维断裂紊乱,出现明显损伤,且LncRNA p21、MMP-2、MMP-9、caspase-3表达和细胞凋亡率均增加(P<0.01),TIMP-1、cyclinD1和YAP、TAZ表达均降低(P<0.01)。与model组相比,sh-LncRNA p21组小鼠血管弹力纤维断裂及损伤程度减轻,LncRNA p21、MMP-2、MMP-9、caspase-3表达和细胞凋亡率均降低(P<0.01),TIMP-1、cyclinD1和YAP、TAZ表达均增加(P<0.01)。结论 LncRNA p21能够促进小鼠AAA形成,其机制与调控Hippo-YAP信号、促进细胞凋亡、抑制细胞增殖相关。

Aim To investigate the effect of long non-coding RNA p21(LncRNA p21)regulating Hippo-Yes-associated protein(Hippo-YAP)signaling pathway on the formation of abdominal aortic aneurysm(AAA)in mice.M ethods C57BL/6 ApoE-/-mouse model of AAA induced by angiotensin II(Ang II)was established by subcutaneously implanting osmotic micropumps.C57BL/6 ApoE-/-mice were randomly divided into sham operation group(sham),model group(model),LncRNA p21 negative control group(sh-NC),LncRNA p21 knockdown group(sh-LncRNA p21).Hematoxylin-staining(HE)and Verhoeff-Van Gieson(VVG)were used to observe the morphological changes of blood vessels.qRT-PCR was used to detect the expression of LncRNA p21.Western blot was used to detect the expression of MMP-2,MMP-9 and TIMP-1.TUNEL staining was used to detect the apoptosis of smooth muscle cells.Western blot was used to detect the expression of caspase-3,cyclinD1 and Hippo-YAP signaling pathway proteins.Results Compared with the sham group,the vascular elastic fibers were disrupted and damaged,the expressions of LncRNA p21,MMP-2,MMP-9,caspase-3 and rate of apoptosis all increased(P<0.01),and the expressions of TIMP-1,cyclinD1 and YAP,TAZ decreased in the model group(P<0.01).Compared with the model group,the vascular elastic fibers were less broken and damaged,and the expressions of LncRNA p21,MMP-2,MMP-9,caspase-3 and the rate of apoptosis all decreased(P<0.01),both TIMP-1,cyclinD1 and YAP,TAZ expressions were raised in the sh-LncRNA p21 group(P<0.01).Conclusions LncRNA p21 can promote the formation of AAA in mice,and its mechanism is related to regulating Hippo-YAP signal,promoting cell apoptosis and inhibiting cell proliferation.