中华实验外科杂志
中華實驗外科雜誌
중화실험외과잡지
CHINESE JOURNAL OF EXPERIMENTAL SURGERY
2013年
5期
921-924
,共4页
黄水传%许托%陈小东%李建文%王三明%张远起%黄胜超
黃水傳%許託%陳小東%李建文%王三明%張遠起%黃勝超
황수전%허탁%진소동%리건문%왕삼명%장원기%황성초
S期激酶相关蛋白2%动脉损伤%血管平滑肌细胞%内膜增生
S期激酶相關蛋白2%動脈損傷%血管平滑肌細胞%內膜增生
S기격매상관단백2%동맥손상%혈관평활기세포%내막증생
S-phase kinase-associated protein 2%Artery injury%Vascular smooth muscle cell%Neointimal hyperplasia
目的 观察S期激酶相关蛋白2(Skp2)在大鼠主动脉球囊损伤后的表达及其对血管内膜增生的影响.方法 将雄性SD大鼠56只随机分为假手术组(n=8)和球囊损伤组(包括2、4、7、10、14、28 d亚组n=8),建立主动脉损伤模型,各组取材后观察血管形态学改变,免疫组织化学链霉菌抗生物素蛋白-过氧化物酶(SP)法检测增殖细胞核抗原(PCNA)、Skp2、细胞周期蛋白依赖性激酶抑制蛋白27(p27KiP1)蛋白表达,实时定量聚合酶链反应(Real-time PCR)检测Skp2和p27Kip1mRNA表达.结果 (1)动脉损伤术后2 d内膜开始出现PCNA阳性的血管平滑肌细胞(VSMC)表达,至7 d达高峰(38.46 ±2.01,P＜ 0.05),此后表达逐渐下降；(2)正常动脉中Skp2阳性表达极低,术后2d开始升高(27.90±2.37),至10d达高峰(47.36±2.37,P＜ 0.05),14 d迅速回落,28 d降至最低；(3)正常动脉中p27Kip1阳性细胞表达明显(27.82 ±3.26),术后2d明显下降(14.71±1.37),10 d时表达迅速增加(32.06±2.57,P＜ 0.05),此后维持在较高水平；(4) Skp2和PCNA表达呈明显正相关(r=0.892,P＜ 0.01)；(5) PCR结果显示正常动脉Skp2 mRNA表达极低(1.28±1.11),术后2d表达开始升高,至10d达高峰(10.49±3.17,P＜ 0.05),此后表达开始下降；正常动脉中可见p27Kip1 mRNA表达(1.14±0.64),术后4d明显下降,7～10d表达仍较低,14d迅速升高(1.64±0.45,P＜ 0.05),至28 d恢复至正常水平.结论 Skp2是VSMC增殖及血管内膜增生的重要促进因子之一.
目的 觀察S期激酶相關蛋白2(Skp2)在大鼠主動脈毬囊損傷後的錶達及其對血管內膜增生的影響.方法 將雄性SD大鼠56隻隨機分為假手術組(n=8)和毬囊損傷組(包括2、4、7、10、14、28 d亞組n=8),建立主動脈損傷模型,各組取材後觀察血管形態學改變,免疫組織化學鏈黴菌抗生物素蛋白-過氧化物酶(SP)法檢測增殖細胞覈抗原(PCNA)、Skp2、細胞週期蛋白依賴性激酶抑製蛋白27(p27KiP1)蛋白錶達,實時定量聚閤酶鏈反應(Real-time PCR)檢測Skp2和p27Kip1mRNA錶達.結果 (1)動脈損傷術後2 d內膜開始齣現PCNA暘性的血管平滑肌細胞(VSMC)錶達,至7 d達高峰(38.46 ±2.01,P＜ 0.05),此後錶達逐漸下降；(2)正常動脈中Skp2暘性錶達極低,術後2d開始升高(27.90±2.37),至10d達高峰(47.36±2.37,P＜ 0.05),14 d迅速迴落,28 d降至最低；(3)正常動脈中p27Kip1暘性細胞錶達明顯(27.82 ±3.26),術後2d明顯下降(14.71±1.37),10 d時錶達迅速增加(32.06±2.57,P＜ 0.05),此後維持在較高水平；(4) Skp2和PCNA錶達呈明顯正相關(r=0.892,P＜ 0.01)；(5) PCR結果顯示正常動脈Skp2 mRNA錶達極低(1.28±1.11),術後2d錶達開始升高,至10d達高峰(10.49±3.17,P＜ 0.05),此後錶達開始下降；正常動脈中可見p27Kip1 mRNA錶達(1.14±0.64),術後4d明顯下降,7～10d錶達仍較低,14d迅速升高(1.64±0.45,P＜ 0.05),至28 d恢複至正常水平.結論 Skp2是VSMC增殖及血管內膜增生的重要促進因子之一.
목적 관찰S기격매상관단백2(Skp2)재대서주동맥구낭손상후적표체급기대혈관내막증생적영향.방법 장웅성SD대서56지수궤분위가수술조(n=8)화구낭손상조(포괄2、4、7、10、14、28 d아조n=8),건립주동맥손상모형,각조취재후관찰혈관형태학개변,면역조직화학련매균항생물소단백-과양화물매(SP)법검측증식세포핵항원(PCNA)、Skp2、세포주기단백의뢰성격매억제단백27(p27KiP1)단백표체,실시정량취합매련반응(Real-time PCR)검측Skp2화p27Kip1mRNA표체.결과 (1)동맥손상술후2 d내막개시출현PCNA양성적혈관평활기세포(VSMC)표체,지7 d체고봉(38.46 ±2.01,P＜ 0.05),차후표체축점하강；(2)정상동맥중Skp2양성표체겁저,술후2d개시승고(27.90±2.37),지10d체고봉(47.36±2.37,P＜ 0.05),14 d신속회락,28 d강지최저；(3)정상동맥중p27Kip1양성세포표체명현(27.82 ±3.26),술후2d명현하강(14.71±1.37),10 d시표체신속증가(32.06±2.57,P＜ 0.05),차후유지재교고수평；(4) Skp2화PCNA표체정명현정상관(r=0.892,P＜ 0.01)；(5) PCR결과현시정상동맥Skp2 mRNA표체겁저(1.28±1.11),술후2d표체개시승고,지10d체고봉(10.49±3.17,P＜ 0.05),차후표체개시하강；정상동맥중가견p27Kip1 mRNA표체(1.14±0.64),술후4d명현하강,7～10d표체잉교저,14d신속승고(1.64±0.45,P＜ 0.05),지28 d회복지정상수평.결론 Skp2시VSMC증식급혈관내막증생적중요촉진인자지일.
Objective To investigate the change of S-phase kinase-associated protein-2 (Skp2) in the aorta of rats after balloon injury and its effect on neointimal hyperplasia.Methods Fisty-six SD male rats were randomly divided into two groups:sham operated group (n =8),and balloon injured group (including:2-,4-,7-,10-,14-and 28-day subgroups,n =8 each).The model of rat aorta injury was established.The vascular tissues were harvested on the postoperative day (POD) 2,4,7,10,14 and 28 respectively and examined histomorphologically.Inmunohistochemistry streptavid-in-peroxidase (SP) method was used to detect the differential expression level of proliferating cell nuclear antigen (PCNA),Skp2 and cyclin dependent kinase inhibitor protein 27 (p27KiP1) at each time point.The expression levels of Skp2 and p27Kip1 mRNA were detected by using real-time quantitative polymerase chain reaction (Real-time PCR).Results (1) The normal artery wall didn' t express PCNA,hut PCNA was detected on POD 2.The amount of positive cells was increased and reached a peak on POD 7 [(38.46 ± 2.01),P ＜ 0.05],then gradually decreased; (2) The expression of Skp2 was very low in uninjured aorta,and increased on POD 2 [(27.90 ±2.37)] and peaked on POD 10 [(47.36 ±2.37),P ＜ 0.05].It was declined on POD 14 and further decreased on POD 28; (3) p27Kip1 was expressed in normal aorta (27.82 ±3.26) and significantly reduced on POD 2 [(14.71 ± 1.37)],significantly increased on POD 10 [(32.06 ± 2.57),P ＜ 0.05] and remained high until POD 28; (4) The expression of PCNA was temporally associated with expression of Skp2 (r =0.892,P ＜ 0.01) ; (5) The Real-time PCR results showed that the expression of Skp2 mRNA was extremely low in normal aorta [(1.28 ± 1.11)],increased on POD 2 and reached the peak on POD 10 [(10.49 ±3.17),P＜ 0.05],then decreased turther on POD 28.The expression of p27Kip1 mRNA could be detected in normal aorta (1.14 ±0.64),decreased on POD 4 (P ＜ 0.05),and remained low from POD 7 to 10,then significantly increased on POD 14 [(1.64 ±0.45),P ＜ 0.05] and returned to normal level on POD 28 (P ＞ 0.05).Conclusion Skp2 was one of the important promoting factors that induced VSMC proliferation and subsequent neointimal hyperplasia.
