中国医药导报
中國醫藥導報
중국의약도보
CHINA MEDICAL HERALD
2015年
22期
27-32
,共6页
黄进启%姚元波%向水%蔡彦力%郑勇%刘金平
黃進啟%姚元波%嚮水%蔡彥力%鄭勇%劉金平
황진계%요원파%향수%채언력%정용%류금평
细胞因子信号转导抑制剂3%信号转导与转录激活因子%炎症%静脉移植物
細胞因子信號轉導抑製劑3%信號轉導與轉錄激活因子%炎癥%靜脈移植物
세포인자신호전도억제제3%신호전도여전록격활인자%염증%정맥이식물
Suppressor of cytokine signaling 3%Signal transducer and activator of transcription%Inflammation%Vein graft
目的：检测细胞因子信号转导抑制剂3(SOCS3)在大鼠颈外静脉颈总动脉移植物和体外培养的动脉血管平滑肌细胞增殖模型中的表达，探讨其在静脉移植物再狭窄病理过程中的可能作用及机制。方法体内实验分两组，将36只雄性SD大鼠随机分成实验组和对照组，每组18只。实验组采用标准显微外科技术行颈外静脉颈总动脉翻转端吻合，对照组行假手术，分别于术后第1、3、7天取出移植静脉。体外培养的大鼠血管平滑肌细胞用碱性成纤维细胞生长因子(bFGF)刺激，再用大鼠SOCS3基因重组腺病毒(pYrAd-rSOCS3)、对照病毒pYrAd-GFP转染，体外实验分四组：对照组，bFGF组，bFGF+pYrAd-GFP组，bFGF+pYrAd-rSOCS3组。应用Real time-PCR和Western boltting检测白介素6(IL-6)、单核细胞趋化蛋白1(MCP-1)、肿瘤坏死因子α(TNF-α)、白介素1β(IL-1β)、细胞间黏附分子1(ICAM-1)、信号转导与转录激活因子(STAT3)、磷酸化的信号转导与转录激活因子(P-STAT3)、SOCS3 mRNA和蛋白表达水平。结果体内实验表明，与对照组比较，实验组静脉移植物中IL-6[(3.60±0.51)比(1.00±0.00)；(1.52±0.37)比(0.35±0.05)]、MCP-1[(2.08±0.38)比(1.00±0.00)；(1.90±0.31)比(0.85±0.17)]、TNF-α[(4.86±0.74)比(1.00±0.00)；(1.66±0.30)比(0.29±0.07)]、IL-1β[(2.73±0.52)比(1.00±0.00)；(0.74±0.17)比(0.19±0.04)]、ICAM-1[(1.97±0.35)比(1.00±0.00)；(1.02±0.39)比(0.21±0.02)]、SOCS3[(1.93±0.38)比(1.00±0.00)；(0.82±0.18)比(0.42±0.12)]mRNA和蛋白表达水平以及STAT3[(1.50±0.36)比(0.21±0.05)]、P-STAT3[(1.54±0.39)比(0.37±0.10)]蛋白表达水平均在1周内明显升高(P<0.05或P<0.01)。体外实验结果表明，与对照组比较，bFGF组中上述指标mRNA和蛋白表达明显上调(P<0.05或P<0.01)；与bFGF组比较，bFGF+pYrAd-rSOCS3组中SOCS3 mRNA和蛋白[(5.47±1.03)比(1.37±0.24)；(1.79±0.38)比(1.28±0.32)]表达进一步上调(P<0.01、P<0.05)，而IL-6[(1.28±0.25)比(1.57±0.31)；(1.68±0.39)比(2.36±0.48)]、MCP-1[(1.17±0.23)比(2.08±0.37)；(1.25±0.21)比(1.66±0.43)]、TNF-α[(1.37±0.23)比(3.06±0.52)；(1.20±0.24)比(1.54±0.31)]、IL-1β[(1.48±0.21)比(1.71±0.19)；(1.00±0.24)比(1.49±0.35)]、ICAM-1[(1.34±0.21)比(2.10±0.27)；(0.99±0.21)比(1.41±0.32)]mRNA和蛋白表达水平以及STAT3[(0.77±0.13)比(1.30±0.27)]、P-STAT3[(1.18±0.36)比(1.74±0.36)]蛋白表达明显下调(P<0.05或P<0.01)。结论 SOCS3在静脉移植物病变的早期病理炎性反应中，可能通过抑制其下游信号通路的关键转录因子STAT3的激活及其磷酸化而发挥负向调节作用，可为冠状动脉旁路移植术后静脉移植物再狭窄的理论研究和临床防治提供一种新思路和新靶点。
目的：檢測細胞因子信號轉導抑製劑3(SOCS3)在大鼠頸外靜脈頸總動脈移植物和體外培養的動脈血管平滑肌細胞增殖模型中的錶達，探討其在靜脈移植物再狹窄病理過程中的可能作用及機製。方法體內實驗分兩組，將36隻雄性SD大鼠隨機分成實驗組和對照組，每組18隻。實驗組採用標準顯微外科技術行頸外靜脈頸總動脈翻轉耑吻閤，對照組行假手術，分彆于術後第1、3、7天取齣移植靜脈。體外培養的大鼠血管平滑肌細胞用堿性成纖維細胞生長因子(bFGF)刺激，再用大鼠SOCS3基因重組腺病毒(pYrAd-rSOCS3)、對照病毒pYrAd-GFP轉染，體外實驗分四組：對照組，bFGF組，bFGF+pYrAd-GFP組，bFGF+pYrAd-rSOCS3組。應用Real time-PCR和Western boltting檢測白介素6(IL-6)、單覈細胞趨化蛋白1(MCP-1)、腫瘤壞死因子α(TNF-α)、白介素1β(IL-1β)、細胞間黏附分子1(ICAM-1)、信號轉導與轉錄激活因子(STAT3)、燐痠化的信號轉導與轉錄激活因子(P-STAT3)、SOCS3 mRNA和蛋白錶達水平。結果體內實驗錶明，與對照組比較，實驗組靜脈移植物中IL-6[(3.60±0.51)比(1.00±0.00)；(1.52±0.37)比(0.35±0.05)]、MCP-1[(2.08±0.38)比(1.00±0.00)；(1.90±0.31)比(0.85±0.17)]、TNF-α[(4.86±0.74)比(1.00±0.00)；(1.66±0.30)比(0.29±0.07)]、IL-1β[(2.73±0.52)比(1.00±0.00)；(0.74±0.17)比(0.19±0.04)]、ICAM-1[(1.97±0.35)比(1.00±0.00)；(1.02±0.39)比(0.21±0.02)]、SOCS3[(1.93±0.38)比(1.00±0.00)；(0.82±0.18)比(0.42±0.12)]mRNA和蛋白錶達水平以及STAT3[(1.50±0.36)比(0.21±0.05)]、P-STAT3[(1.54±0.39)比(0.37±0.10)]蛋白錶達水平均在1週內明顯升高(P<0.05或P<0.01)。體外實驗結果錶明，與對照組比較，bFGF組中上述指標mRNA和蛋白錶達明顯上調(P<0.05或P<0.01)；與bFGF組比較，bFGF+pYrAd-rSOCS3組中SOCS3 mRNA和蛋白[(5.47±1.03)比(1.37±0.24)；(1.79±0.38)比(1.28±0.32)]錶達進一步上調(P<0.01、P<0.05)，而IL-6[(1.28±0.25)比(1.57±0.31)；(1.68±0.39)比(2.36±0.48)]、MCP-1[(1.17±0.23)比(2.08±0.37)；(1.25±0.21)比(1.66±0.43)]、TNF-α[(1.37±0.23)比(3.06±0.52)；(1.20±0.24)比(1.54±0.31)]、IL-1β[(1.48±0.21)比(1.71±0.19)；(1.00±0.24)比(1.49±0.35)]、ICAM-1[(1.34±0.21)比(2.10±0.27)；(0.99±0.21)比(1.41±0.32)]mRNA和蛋白錶達水平以及STAT3[(0.77±0.13)比(1.30±0.27)]、P-STAT3[(1.18±0.36)比(1.74±0.36)]蛋白錶達明顯下調(P<0.05或P<0.01)。結論 SOCS3在靜脈移植物病變的早期病理炎性反應中，可能通過抑製其下遊信號通路的關鍵轉錄因子STAT3的激活及其燐痠化而髮揮負嚮調節作用，可為冠狀動脈徬路移植術後靜脈移植物再狹窄的理論研究和臨床防治提供一種新思路和新靶點。
목적：검측세포인자신호전도억제제3(SOCS3)재대서경외정맥경총동맥이식물화체외배양적동맥혈관평활기세포증식모형중적표체，탐토기재정맥이식물재협착병리과정중적가능작용급궤제。방법체내실험분량조，장36지웅성SD대서수궤분성실험조화대조조，매조18지。실험조채용표준현미외과기술행경외정맥경총동맥번전단문합，대조조행가수술，분별우술후제1、3、7천취출이식정맥。체외배양적대서혈관평활기세포용감성성섬유세포생장인자(bFGF)자격，재용대서SOCS3기인중조선병독(pYrAd-rSOCS3)、대조병독pYrAd-GFP전염，체외실험분사조：대조조，bFGF조，bFGF+pYrAd-GFP조，bFGF+pYrAd-rSOCS3조。응용Real time-PCR화Western boltting검측백개소6(IL-6)、단핵세포추화단백1(MCP-1)、종류배사인자α(TNF-α)、백개소1β(IL-1β)、세포간점부분자1(ICAM-1)、신호전도여전록격활인자(STAT3)、린산화적신호전도여전록격활인자(P-STAT3)、SOCS3 mRNA화단백표체수평。결과체내실험표명，여대조조비교，실험조정맥이식물중IL-6[(3.60±0.51)비(1.00±0.00)；(1.52±0.37)비(0.35±0.05)]、MCP-1[(2.08±0.38)비(1.00±0.00)；(1.90±0.31)비(0.85±0.17)]、TNF-α[(4.86±0.74)비(1.00±0.00)；(1.66±0.30)비(0.29±0.07)]、IL-1β[(2.73±0.52)비(1.00±0.00)；(0.74±0.17)비(0.19±0.04)]、ICAM-1[(1.97±0.35)비(1.00±0.00)；(1.02±0.39)비(0.21±0.02)]、SOCS3[(1.93±0.38)비(1.00±0.00)；(0.82±0.18)비(0.42±0.12)]mRNA화단백표체수평이급STAT3[(1.50±0.36)비(0.21±0.05)]、P-STAT3[(1.54±0.39)비(0.37±0.10)]단백표체수평균재1주내명현승고(P<0.05혹P<0.01)。체외실험결과표명，여대조조비교，bFGF조중상술지표mRNA화단백표체명현상조(P<0.05혹P<0.01)；여bFGF조비교，bFGF+pYrAd-rSOCS3조중SOCS3 mRNA화단백[(5.47±1.03)비(1.37±0.24)；(1.79±0.38)비(1.28±0.32)]표체진일보상조(P<0.01、P<0.05)，이IL-6[(1.28±0.25)비(1.57±0.31)；(1.68±0.39)비(2.36±0.48)]、MCP-1[(1.17±0.23)비(2.08±0.37)；(1.25±0.21)비(1.66±0.43)]、TNF-α[(1.37±0.23)비(3.06±0.52)；(1.20±0.24)비(1.54±0.31)]、IL-1β[(1.48±0.21)비(1.71±0.19)；(1.00±0.24)비(1.49±0.35)]、ICAM-1[(1.34±0.21)비(2.10±0.27)；(0.99±0.21)비(1.41±0.32)]mRNA화단백표체수평이급STAT3[(0.77±0.13)비(1.30±0.27)]、P-STAT3[(1.18±0.36)비(1.74±0.36)]단백표체명현하조(P<0.05혹P<0.01)。결론 SOCS3재정맥이식물병변적조기병리염성반응중，가능통과억제기하유신호통로적관건전록인자STAT3적격활급기린산화이발휘부향조절작용，가위관상동맥방로이식술후정맥이식물재협착적이론연구화림상방치제공일충신사로화신파점。
Objective To detect the expression of suppressor of cytokine signaling 3 (SOCS3) in vein grafts of rat and explore its possible effect and mechanism in vein graft stenosis. Methods The experiment in vivo was divided into two groups. Thirty-six male Sprague-Dawley rats were randomly divided into two groups: experimental group and control group, with 18 cases in each group. Autologous jugular vein to carotid artery reverse interposition grafts were performed by standard microsurgical tech-nique in rats of experimental group. Sham operation was performed in rats of control group. The vein grafts were har-vested on day 1, day 3, and day 7 after operation. The rat vascular smooth muscle cells were stimulated with basic fi-broblast growth factor (bFGF) and then transfected with the recombinant adenovirus carrying the rat SOCS3 gene (pYrAd-rSOCS3), the matched control adenovirus vector (pYrAd-GFP), which carries only GFP. Experiments included the following groups: control group, bFGF group, bFGF+pYrAd-GFP group, bFGF+pYrAd-rSOCS3 group. The harvest-ed vein grafts and VSMCs were analyzed by real time-PCR and Western blot to detect the mRNA and protein expres-sion levels of interleukin 6 (IL-6), monocyte chemotactic protein (MCP)-1, tumor necrosis factor (TNF)-α, interleukin-1β (IL-1β), intercellular adhension molecular (ICAM)-1, signal transducer and activator of transcription (STAT3), P-STAT3, SOCS3. Results In v iv o, compared with control group, the mRNA and protein expression levels of IL-6 [(3.60±0.51) vs (1.00±0.00); (1.52±0.37) vs (0.35±0.05)], MCP-1 [(2.08±0.38) vs (1.00±0.00); (1.90±0.31) vs (0.85±0.17)], TNF-α [(4.86±0.74) vs (1.00±0.00); (1.66±0.30) vs (0.29±0.07)], IL-1β [(2.73±0.52) vs (1.00±0.00); (0.74±0.17) vs (0.19±0.04)], ICAM-1 [(1.97±0.35) vs (1.00±0.00); (1.02±0.39) vs (0.21±0.02)], SOCS3 [(1.93±0.38) vs (1.00±0.00);(0.82±0.18) v s (0.42±0.12)] and protein expression levels of STAT3 [(1.50±0.36) v s (0.21±0.05)] and P-STAT3 [(1.54±0.39) vs (0.37±0.10)] were significantly higher in the graft samples within 1 week after operation (P<0.05 or P<0.01). In v itro, the expression of above cytokines were increased in bFGF-induced group compared with the control group (P<0.05 or P<0.01). Compared with bFGF group, the mRNA and protein expression of SOCS3 increased [(5.47±1.03) v s (1.37±0.24);(1.79±0.38) v s (1.28±0.32)] (P<0.01, P<0.05), whereas the mRNA and protein expression levels of IL-6 [(1.28±0.25) vs (1.57±0.31); (1.68±0.39) vs (2.36±0.48)], MCP-1 [(1.17±0.23) vs (2.08±0.37); (1.25±0.21) vs (1.66±0.43)], TNF-α[(1.37±0.23) v s (3.06±0.52);(1.20±0.24) v s (1.54±0.31)], IL-1β[(1.48±0.21) v s (1.71±0.19);(1.00±0.24) v s (1.49±0.35)], ICAM-1 [(1.34±0.21) v s (2.10±0.27); (0.99±0.21) v s (1.41±0.32)] and protein expression levels of STAT3 [(0.77±0.13) vs (1.30±0.27), P=0.017] and P-STAT3 [(1.18±0.36) vs (1.74±0.36), P=0.026] were significantly decreased in bFGF+pYrAd-rSOCS3 group (P<0.05 or P<0.01). Conclusion SOCS3 may play a negative regulation through in-hibiting the activation and phosphorylation of key transcription factor STAT3 of downstream signaling pathways during the early inflammatory response in vein graft lesions, which can provide a new thought and a new target for theoretical research and clinical prevention of graft vein stenosis therapy after coronary artery bypass graft.
