CAJ | 학술논문

目的探讨慢病毒重组血管紧张素转换酶(LV-ACE)2基因过表达对于大鼠颈总动脉缺血再灌注损伤(IRI)后内膜新生的影响和相关机制.方法将SD大鼠分为正常对照组、假手术组、IRI后1、2、4周亚组,每组10只,采用夹闭颈总动脉方法制造动脉硬化模型,HE染色观察各组新生内膜面积/中膜面积(I/M)值；另选SD大鼠50只,分为5组:对照组、IRI组、IRI+慢病毒重组绿色荧光蛋白(LV-GFP)组(GFP组)、IRI+ LV-ACE2组(ACE2组)和IRI+紫杉醇组(紫杉醇组),每组10只,夹闭法后利用LV-ACE2干预,并用LV-GFP、紫杉醇等作为对照.HE染色观察各组I/M值,免疫组织化学方法观察各组新生内膜中ACE2、平滑肌细胞α-肌动蛋白(α-SM-actin)、血小板内皮细胞黏附分子(CD31)、血管紧张素1型受体(ATI R)、细胞外信号调节激酶磷酸化(P-ERK1/2)表达变化.结果 (1)IRI后,HE染色观察损伤组的1、2、4周亚组I/M值(分别为0.364±0.032、0.789±0.120和1.517±0.151)均高于正常组(0.011 ±0.004,P均＜0.01).(2)LV-ACE2等干预后,HE染色观察ACE2组I/M值低于IRI组(0.71 ±0.17比1.53 ±0.16,P＜0.01)；免疫组织化学观察ACE2组ACE2表达积分吸光度(IA)值与IRI组比较,差异无统计学意义(1294±283比1080±252,P=0.63),ACE2组CD31表达血管密度低于IRI组[(12.40±4.21)个/mm2比(96.20±17.79)个/mm2,P＜0.01],ACE2组α-SM-actin、AT1R、P-ERK1/2表达IA值均低于IRI组(分别为5 843±839比12 648±1 760,1 219±175比4 861±545,1 040 ±215比2 938±286,P均＜0.01)结论大鼠颈总动脉IRI后4周能够形成稳定的内膜新生,ACE2能明显抑制IRI后内膜新生、新生内膜中平滑肌细胞增殖及新生血管生成,其机制可能与ACE2抑制AT1 R、P-ERK1/2的表达有关.
목적 탐토만병독중조혈관긴장소전환매(LV-ACE)2기인과표체대우대서경총동맥결혈재관주손상(IRI)후내막신생적영향화상관궤제.방법 장SD대서분위정상대조조、가수술조、IRI후1、2、4주아조,매조10지,채용협폐경총동맥방법제조동맥경화모형,HE염색관찰각조신생내막면적/중막면적(I/M)치；령선SD대서50지,분위5조:대조조、IRI조、IRI+만병독중조록색형광단백(LV-GFP)조(GFP조)、IRI+ LV-ACE2조(ACE2조)화IRI+자삼순조(자삼순조),매조10지,협폐법후이용LV-ACE2간예,병용LV-GFP、자삼순등작위대조.HE염색관찰각조I/M치,면역조직화학방법관찰각조신생내막중ACE2、평활기세포α-기동단백(α-SM-actin)、혈소판내피세포점부분자(CD31)、혈관긴장소1형수체(ATI R)、세포외신호조절격매린산화(P-ERK1/2)표체변화.결과 (1)IRI후,HE염색관찰손상조적1、2、4주아조I/M치(분별위0.364±0.032、0.789±0.120화1.517±0.151)균고우정상조(0.011 ±0.004,P균＜0.01).(2)LV-ACE2등간예후,HE염색관찰ACE2조I/M치저우IRI조(0.71 ±0.17비1.53 ±0.16,P＜0.01)；면역조직화학관찰ACE2조ACE2표체적분흡광도(IA)치여IRI조비교,차이무통계학의의(1294±283비1080±252,P=0.63),ACE2조CD31표체혈관밀도저우IRI조[(12.40±4.21)개/mm2비(96.20±17.79)개/mm2,P＜0.01],ACE2조α-SM-actin、AT1R、P-ERK1/2표체IA치균저우IRI조(분별위5 843±839비12 648±1 760,1 219±175비4 861±545,1 040 ±215비2 938±286,P균＜0.01)결론 대서경총동맥IRI후4주능구형성은정적내막신생,ACE2능명현억제IRI후내막신생、신생내막중평활기세포증식급신생혈관생성,기궤제가능여ACE2억제AT1 R、P-ERK1/2적표체유관.
Objective To explore the effects of lentiviral recombinant angiotensin-converting enzyme 2 (LV-ACE2) gene transfer on the neointimal formation after carotid artery ischemia-reperfusion injury (IRI) and related mechanisms.Methods IRI was induced in SD rats through the carotid artery clipping and rats were divided into IRI,IRI + LV-GFP,IRI + LV-ACE2,IRI + paclitaxel groups (n =10each).Sham operated rats serve as normal control.Four weeks later,neointimal formation was observed on HE stained carotid artery sections.The protein expression of ACE2,α-SM-actin,CD31,AT1R and P-ERK were detected by immunohistochemistry.Results (1) Carotid artery neointimal hyperplasia was readily shown in IRI group [I/M:1.517 ± 0.151 (4 weeks later) vs.0.011 ± 0.004 (Sham),P ＜ 0.01],which was significantly reduced in IRI + LV-ACE2 (0.71 ± 0.17) and IRI + paclitaxel (0.89 ± 0.21) groups.(2) The growth of vascular smooth muscle cells and neovascularization were also significantly inhibited in IRI + LV-ACE2 group and the expression of α-SM-actin (5 843 ± 839 vs.12 648 ± 1 760,P ＜ 0.01) and CD31 [(12.40±4.01)/mm2vs.(96.20±17.79)/mm2,P＜0.01],AT1R (1 219 ±175 vs.4 861±545,P ＜0.01) and P-ERK1/2 phosphorylation (1 040 ±215 vs.2 938 ±286,P ＜0.01) in the neointimal of the injury arteries in IRI + LV-ACE2 group were significantly downregulated compared to IRI group.Conclusion This data suggest that ACE2 gene overexpression is able to attenuate neointimal formation after ischemia-reperfusion injury possibly through downregulating AT1 receptor expression and signal pathway of ERK1/2 phosphorylation.