中华心血管病杂志
中華心血管病雜誌
중화심혈관병잡지
Chinese Journal of Cardiology
2010年
6期
531-538
,共8页
曾武涛%陈伟燕%冷秀玉%何建桂%马虹
曾武濤%陳偉燕%冷秀玉%何建桂%馬虹
증무도%진위연%랭수옥%하건계%마홍
血管紧张素类%胶原%信号传导
血管緊張素類%膠原%信號傳導
혈관긴장소류%효원%신호전도
Angiotensin%Collagen%Signal transduction
目的 探讨血管紧张素(ANG)-(1-7)对兔腹主动脉球囊损伤后血管壁胶原合成的影响及转化生长因子(TGF)β/Smad通路在其中的作用.方法 将32只健康新西兰白兔随机分为4组.假手术组不予任何处理,模型组、ANG-(1-7)组和ANG-(1-7)+A-779组均行腹主动脉球囊扩张术,术后分别通过微泵持续静脉给予生理盐水(2.5 μl/h)、ANG-(1-7)(576 μg·kg~(-1)·d~(-1)及ANG-(1-7)+A-779(576 μg·kg~(-1·d~(-1)4周.于术前、术后4周采血,用酶联免疫吸附测定方法测定血浆中ANG Ⅱ的水平.术后4周行腹主动脉造影及苏木精-伊红染色,计算血管腔的内径及新生内膜面积和管腔狭窄率等.以逆转录聚合酶链反应及免疫组织化学法测定血管壁中Ⅰ、Ⅲ型胶原蛋白的含量和表达部位,同时以Western blot方法测定血管组织中TGF-β_1、磷酸化Smad2的表达水平.结果 术后4周,与模型组相比,ANG-(1-7)组和ANG-(1-7)+A-779组管腔内径较大[(4.11 ±0.10)mm和(3.34±0.1) mm比(2.88 ±0.08)mm,P均<0.05],新生内膜面积[(0.27±0.09)mm~2和(0.38±0.01)mm~2比(0.41±0.02)mm~2,P均<0.05],内膜厚度[(208 ±17)μm和(407 ±25)μm比(448 ±15)μm,P均<0.05]和管腔狭窄率[(28.1 ±2.7)%和(36.8 ±2.2)%比(40.1 ±2.7)%,P均<0.05]均较小,ANG-(1-7)+A-779组与ANG-(1-7)组间差异也有统计学意义(P<0.05).与假手术组比较,术后4周模型组、ANG-(1-7)组和ANG-(1-7)+A-779组血管组织中Ⅰ、Ⅲ型胶原蛋白,TGF-β_1和磷酸化Smad2的表达水平均较高(P<0.01或P<0.05),ANG-(1-7)组Ⅰ、Ⅲ型胶原蛋白,TGF-β_1和磷酸化Smad2的表达均低于模型组(P均<0.05),ANG-(1-7)+A-779组与模型组间Ⅰ、Ⅲ型胶原蛋白,TGF-β_1和磷酸化Smad2的表达差异无统计学意义.术后模型组和ANG-(1-7)组血浆中ANG Ⅱ水平均较术前升高(P<0.05),但两组差异无统计学意义.结论 ANG-(1-7)可明显减轻兔腹主动脉球囊损伤后的再狭窄及血管壁胶原合成,这可能与调节组织中TGF-β_1的表达,间接抑制Smads途径的激活有关.
目的 探討血管緊張素(ANG)-(1-7)對兔腹主動脈毬囊損傷後血管壁膠原閤成的影響及轉化生長因子(TGF)β/Smad通路在其中的作用.方法 將32隻健康新西蘭白兔隨機分為4組.假手術組不予任何處理,模型組、ANG-(1-7)組和ANG-(1-7)+A-779組均行腹主動脈毬囊擴張術,術後分彆通過微泵持續靜脈給予生理鹽水(2.5 μl/h)、ANG-(1-7)(576 μg·kg~(-1)·d~(-1)及ANG-(1-7)+A-779(576 μg·kg~(-1·d~(-1)4週.于術前、術後4週採血,用酶聯免疫吸附測定方法測定血漿中ANG Ⅱ的水平.術後4週行腹主動脈造影及囌木精-伊紅染色,計算血管腔的內徑及新生內膜麵積和管腔狹窄率等.以逆轉錄聚閤酶鏈反應及免疫組織化學法測定血管壁中Ⅰ、Ⅲ型膠原蛋白的含量和錶達部位,同時以Western blot方法測定血管組織中TGF-β_1、燐痠化Smad2的錶達水平.結果 術後4週,與模型組相比,ANG-(1-7)組和ANG-(1-7)+A-779組管腔內徑較大[(4.11 ±0.10)mm和(3.34±0.1) mm比(2.88 ±0.08)mm,P均<0.05],新生內膜麵積[(0.27±0.09)mm~2和(0.38±0.01)mm~2比(0.41±0.02)mm~2,P均<0.05],內膜厚度[(208 ±17)μm和(407 ±25)μm比(448 ±15)μm,P均<0.05]和管腔狹窄率[(28.1 ±2.7)%和(36.8 ±2.2)%比(40.1 ±2.7)%,P均<0.05]均較小,ANG-(1-7)+A-779組與ANG-(1-7)組間差異也有統計學意義(P<0.05).與假手術組比較,術後4週模型組、ANG-(1-7)組和ANG-(1-7)+A-779組血管組織中Ⅰ、Ⅲ型膠原蛋白,TGF-β_1和燐痠化Smad2的錶達水平均較高(P<0.01或P<0.05),ANG-(1-7)組Ⅰ、Ⅲ型膠原蛋白,TGF-β_1和燐痠化Smad2的錶達均低于模型組(P均<0.05),ANG-(1-7)+A-779組與模型組間Ⅰ、Ⅲ型膠原蛋白,TGF-β_1和燐痠化Smad2的錶達差異無統計學意義.術後模型組和ANG-(1-7)組血漿中ANG Ⅱ水平均較術前升高(P<0.05),但兩組差異無統計學意義.結論 ANG-(1-7)可明顯減輕兔腹主動脈毬囊損傷後的再狹窄及血管壁膠原閤成,這可能與調節組織中TGF-β_1的錶達,間接抑製Smads途徑的激活有關.
목적 탐토혈관긴장소(ANG)-(1-7)대토복주동맥구낭손상후혈관벽효원합성적영향급전화생장인자(TGF)β/Smad통로재기중적작용.방법 장32지건강신서란백토수궤분위4조.가수술조불여임하처리,모형조、ANG-(1-7)조화ANG-(1-7)+A-779조균행복주동맥구낭확장술,술후분별통과미빙지속정맥급여생리염수(2.5 μl/h)、ANG-(1-7)(576 μg·kg~(-1)·d~(-1)급ANG-(1-7)+A-779(576 μg·kg~(-1·d~(-1)4주.우술전、술후4주채혈,용매련면역흡부측정방법측정혈장중ANG Ⅱ적수평.술후4주행복주동맥조영급소목정-이홍염색,계산혈관강적내경급신생내막면적화관강협착솔등.이역전록취합매련반응급면역조직화학법측정혈관벽중Ⅰ、Ⅲ형효원단백적함량화표체부위,동시이Western blot방법측정혈관조직중TGF-β_1、린산화Smad2적표체수평.결과 술후4주,여모형조상비,ANG-(1-7)조화ANG-(1-7)+A-779조관강내경교대[(4.11 ±0.10)mm화(3.34±0.1) mm비(2.88 ±0.08)mm,P균<0.05],신생내막면적[(0.27±0.09)mm~2화(0.38±0.01)mm~2비(0.41±0.02)mm~2,P균<0.05],내막후도[(208 ±17)μm화(407 ±25)μm비(448 ±15)μm,P균<0.05]화관강협착솔[(28.1 ±2.7)%화(36.8 ±2.2)%비(40.1 ±2.7)%,P균<0.05]균교소,ANG-(1-7)+A-779조여ANG-(1-7)조간차이야유통계학의의(P<0.05).여가수술조비교,술후4주모형조、ANG-(1-7)조화ANG-(1-7)+A-779조혈관조직중Ⅰ、Ⅲ형효원단백,TGF-β_1화린산화Smad2적표체수평균교고(P<0.01혹P<0.05),ANG-(1-7)조Ⅰ、Ⅲ형효원단백,TGF-β_1화린산화Smad2적표체균저우모형조(P균<0.05),ANG-(1-7)+A-779조여모형조간Ⅰ、Ⅲ형효원단백,TGF-β_1화린산화Smad2적표체차이무통계학의의.술후모형조화ANG-(1-7)조혈장중ANG Ⅱ수평균교술전승고(P<0.05),단량조차이무통계학의의.결론 ANG-(1-7)가명현감경토복주동맥구낭손상후적재협착급혈관벽효원합성,저가능여조절조직중TGF-β_1적표체,간접억제Smads도경적격활유관.
Objective To explore the effects of Angiotensin (ANG)-( 1-7 ) on postangioplasty fibrotic remodeling and the involvement of TGF-β/Smad signaling pathway in this process. Methods Thirty two healthy New Zealand white rabbits were randomly divided into 4 groups: sham group, control group,ANG-(1-7) group and ANG-( 1-7 ) + A-779 group. Rabbits underwent angioplasty in the abdominal aorta or sham surgery. Subsequently, an osmotic minipump was implanted for saline, ANG-(1-7) (576 μg· kg~(-1) ·d~(-1)) or ANG-(1-7) + A-779 (576 μg · kg~(-1) · d~(-1)) delivery. Before and after 4 weeks treatment, the levels of ANG Ⅱ in plasma were measured by ELISA. At week 4, angiography and histomorphometric analysis were performed, mRNA levels of collagen Ⅰ and Ⅲ were assayed by RT-PCR and protein levels of TGF-β_1 and Stand2 in local vessel were assayed by Western blot. Results Following 4 weeks treatment,ANG-( 1-7 ) and ANG-( 1-7 ) + A-779 group displayed a significant elevation in lumen diameter [ (4. 11±0.10) mm and (3.34 ±0. 11) mm vs. (2.88 ±0.08) mm,P <0.05, respectively] and reduction in neointimal thickness [ (208 ± 17) μm and (407±25) μm vs. (448 ± 15) μm, P <0. 05, respectively],neointimal area [(0.27 ±0.09) mm~2 and (0.38 ±0.01) mm~2 vs. (0.41 ±0.02) mm~2, P<0.05,respectively] and restenosis rate [(28. 1 ±2.7)% and (36.8 ±2.2)% vs. (40. 1 ±2.7)% ,P<0.05,respectively] compared with control group. Collagen Ⅰ , Ⅲ mRNA and TGF-β_1 , Smad2 protein levels were significantly elevated in control group,ANG-(1-7) group and ANG-(1-7) + A-779 group compared to sham group ( P < 0. 01 or P < 0. 05 ) and reduced in ANG- (1-7) group compared to control group ( all P < 0. 05).Co-treatment with A-779 reversed the inhibitory action of ANG-( 1-7 ) . Plasma levels of ANG Ⅱ postangioplasty were similar in control and ANG-( 1-7 ) group and both were significantly higher than preoperation levels. Conclusion ANG-(1-7) attenuates postangioplasty collagen synthesis in rabbits possibly through down-regulating the expression of TGF-β_1 and inhibiting the activation of Smad2 pathway.
