CAJ | 학술논문

目的探讨外膜炎症及血清炎症因子与移植物动脉硬化早期发病的关系.方法把36只同种异体胸主动脉腹腔移植大鼠及16只同系移植对照大鼠随机分成4组(每组实验组9只,对照组4只):A组,术后1周处死;B组,术后2周处死;C组,术后3周处死;D组,术后4周处死.术前及处死时抽血分离留取血清,使用酶联免疫吸附法检测血清炎症因子白细胞介素6(interleukin-6,IL-6)和肿瘤坏死因子α(tumor necrosis factor-alpha,TNF-α)水平,处死后留取血管标本,HE染色观察血管外膜炎症细胞浸润程度及病理改变;用免疫组织化学法检测α-肌动蛋白(α-actin)、周期蛋白依赖性激酶1(cyclin dependent kinase-1,CDK1)、增殖细胞核抗原(proliferating cell nuclear antigen,PCNA)在血管壁中的表达.对比各组与术前血清炎症因子水平变化及各组间观察指标的变化.结果术后7 d,外膜大量炎症细胞浸润;术后14 d,外膜有轻度胶原纤维增生伴炎症浸润;术后28 d,外膜明显增厚,内有大量增生的平滑肌细胞、胶原纤维及炎症细胞,血管中层断裂,可见外膜平滑肌细胞迁移至内膜,内膜亦出现明显纤维化及平滑肌细胞增生.在血管外膜平滑肌细胞中,α-肌动蛋白、PCNA和CDK.表达愈来愈高(均P<0.05),且早于内膜.移植术后A、B、C、D组血清炎症介质水平均高于术前基础水平.IL-6水平,B实验组高于术前(P<0.05),A、C、D实验组显著高于术前(均P<0.01),A、B、C对照组也高于术前基础水平(均P<0.05);A、C、D实验组显著高于对照组(均P<0.01).TNF-α水平,A、B、C实验组高于术前(均P<0.05),D实验组显著高于术前(P<0.01),A、B、C、D对照组与术前无明显变化,各实验组均明显高于对照组(均P<0.01),差异有统计学意义.结论大鼠同种异体胸主动脉移植模型建立后,随着外膜炎症加重,移植血管外膜及内膜出现明显动脉硬化;血清炎症介质水平呈持续升高,提示外膜炎症细胞浸润及炎症因子均参与移植物动脉硬化的早期发生. 目的探討外膜炎癥及血清炎癥因子與移植物動脈硬化早期髮病的關繫.方法把36隻同種異體胸主動脈腹腔移植大鼠及16隻同繫移植對照大鼠隨機分成4組(每組實驗組9隻,對照組4隻):A組,術後1週處死;B組,術後2週處死;C組,術後3週處死;D組,術後4週處死.術前及處死時抽血分離留取血清,使用酶聯免疫吸附法檢測血清炎癥因子白細胞介素6(interleukin-6,IL-6)和腫瘤壞死因子α(tumor necrosis factor-alpha,TNF-α)水平,處死後留取血管標本,HE染色觀察血管外膜炎癥細胞浸潤程度及病理改變;用免疫組織化學法檢測α-肌動蛋白(α-actin)、週期蛋白依賴性激酶1(cyclin dependent kinase-1,CDK1)、增殖細胞覈抗原(proliferating cell nuclear antigen,PCNA)在血管壁中的錶達.對比各組與術前血清炎癥因子水平變化及各組間觀察指標的變化.結果術後7 d,外膜大量炎癥細胞浸潤;術後14 d,外膜有輕度膠原纖維增生伴炎癥浸潤;術後28 d,外膜明顯增厚,內有大量增生的平滑肌細胞、膠原纖維及炎癥細胞,血管中層斷裂,可見外膜平滑肌細胞遷移至內膜,內膜亦齣現明顯纖維化及平滑肌細胞增生.在血管外膜平滑肌細胞中,α-肌動蛋白、PCNA和CDK.錶達愈來愈高(均P<0.05),且早于內膜.移植術後A、B、C、D組血清炎癥介質水平均高于術前基礎水平.IL-6水平,B實驗組高于術前(P<0.05),A、C、D實驗組顯著高于術前(均P<0.01),A、B、C對照組也高于術前基礎水平(均P<0.05);A、C、D實驗組顯著高于對照組(均P<0.01).TNF-α水平,A、B、C實驗組高于術前(均P<0.05),D實驗組顯著高于術前(P<0.01),A、B、C、D對照組與術前無明顯變化,各實驗組均明顯高于對照組(均P<0.01),差異有統計學意義.結論大鼠同種異體胸主動脈移植模型建立後,隨著外膜炎癥加重,移植血管外膜及內膜齣現明顯動脈硬化;血清炎癥介質水平呈持續升高,提示外膜炎癥細胞浸潤及炎癥因子均參與移植物動脈硬化的早期髮生.
목적 탐토외막염증급혈청염증인자여이식물동맥경화조기발병적관계.방법 파36지동충이체흉주동맥복강이식대서급16지동계이식대조대서수궤분성4조(매조실험조9지,대조조4지):A조,술후1주처사;B조,술후2주처사;C조,술후3주처사;D조,술후4주처사.술전급처사시추혈분리류취혈청,사용매련면역흡부법검측혈청염증인자백세포개소6(interleukin-6,IL-6)화종류배사인자α(tumor necrosis factor-alpha,TNF-α)수평,처사후류취혈관표본,HE염색관찰혈관외막염증세포침윤정도급병리개변;용면역조직화학법검측α-기동단백(α-actin)、주기단백의뢰성격매1(cyclin dependent kinase-1,CDK1)、증식세포핵항원(proliferating cell nuclear antigen,PCNA)재혈관벽중적표체.대비각조여술전혈청염증인자수평변화급각조간관찰지표적변화.결과 술후7 d,외막대량염증세포침윤;술후14 d,외막유경도효원섬유증생반염증침윤;술후28 d,외막명현증후,내유대량증생적평활기세포、효원섬유급염증세포,혈관중층단렬,가견외막평활기세포천이지내막,내막역출현명현섬유화급평활기세포증생.재혈관외막평활기세포중,α-기동단백、PCNA화CDK.표체유래유고(균P<0.05),차조우내막.이식술후A、B、C、D조혈청염증개질수평균고우술전기출수평.IL-6수평,B실험조고우술전(P<0.05),A、C、D실험조현저고우술전(균P<0.01),A、B、C대조조야고우술전기출수평(균P<0.05);A、C、D실험조현저고우대조조(균P<0.01).TNF-α수평,A、B、C실험조고우술전(균P<0.05),D실험조현저고우술전(P<0.01),A、B、C、D대조조여술전무명현변화,각실험조균명현고우대조조(균P<0.01),차이유통계학의의.결론 대서동충이체흉주동맥이식모형건립후,수착외막염증가중,이식혈관외막급내막출현명현동맥경화;혈청염증개질수평정지속승고,제시외막염증세포침윤급염증인자균삼여이식물동맥경화적조기발생.
Objective To study the roles of serum inflammatory factors IL-6 and TNF-α and allograft adventitial inflammation in the pathogenesis of allograft arteriosclerosis in rats. Methods Thirty-six allogeneic allograft rats and 16 syngeneic allograft rats were randomly divided into 4 groups (9 rats in each experimental group and 4 in each control group): A, harvested at Week 1 post-operation; B, harvested at Week 2 post-operation; C, harvested at Week 3 post-operation; D, harvested at Week 4 post-operation. Blood samples were collected before transplantation and after harvest. The method of ELISA was used for testing serum inflammatory factors including interleukin-6 (IL-6), tumor necrosis factor-α (TNF-α), HE staining for pathologic changes of aortic allograft and immunohistochemical method for expression of α-actin, cyclin dependent kinase-1 (CDK1) and proliferating cell nuclear antigen (PCNA). Compare the inflammatory factors and other observations between groups and preoperative. Results At Week 1 post-operation, a large amount of inflammatory cell infiltration in adventitia was observed; at Week 2 post-operation, slight collagen fibers hyperplasia with inflammatory infiltration; at Week 4 post-operation, obvious odventitia thickening with a large number of smooth muscle cells, collagen fibers and inflammatory cells, smooth muscle cells migration from adventitia to intima. Expressions of α-actin, CDK1 and PCNA kept increasing with time in adventitia (P < 0.05). There was a significant increase in serum TNF-α level in Groups A, B, C and D, as compared with pre-operative basal level (P <0.01). There was no difference between controls and pre-operative basal level. IL-6 level slightly declined in the middle stage, but finally increased in experimental group B (P <0.0.5) while it significantly increased in Groups A, C, D (P <0.01). In the control groups A, B, C, it was higher than pre-operative level (P < 0.05). In experimental groups A, C, D, it had a significant increase as compared with controls (P < 0.01). Conclusions In abdominal aortic allograft models, obvious angiosclerosis was found in adventitia and intima in accordance with the severity of odventitial inflammation. Thus the inflammatory factors and inflammatory cell infiltration in odventitia are both involved in the pathogenesis of early allograft arteriosclerosis.