【作者】 张波；

【导师】 孔宪炳；

【作者基本信息】 重庆医科大学 ， 外科学， 2008， 硕士

【摘要】 目的:检测CO₂气腹下兔肝VX2肿瘤MMP2及CD34的表达,探讨CO₂对肿瘤侵袭生长转移的影响及机制。方法:选用2.5～3.5kg健康雄性新西兰兔,采用组织块移植法建立兔肝VX2肿瘤模型,术后实验动物在同等条件下饲养1周。把种植成功后的30只实验兔随机分为3组:气腹组（A组,n=10）、基质金属蛋白酶抑制剂（MMPi）组（B组,n=10）及免气腹组（C组,n=10）。A组、B组模拟腹腔镜手术,腹腔内注入CO₂气体,压力为7.5mmHg,气体流量为0.4L/min,于右下腹穿刺置入套管针1枚模拟Trocar穿刺,手术开始后A组立即腹腔注射生理盐水5ml,B组立即腹腔注射MMPi,即邻菲罗啉（1,10-Phenanthroline） 25mg/kg,气腹持续3h;C组以2根缝线悬吊腹壁,于右下腹穿刺置入套管针1枚模拟Trocar穿刺,持续3h。所有动物麻醉恢复后,保持在相同饲养条件下,14d后安乐死处死动物,观察肿瘤生长、转移情况,比较各组肿瘤最大径、体积及重量;免疫组织化学方法（SABC法）检测肿瘤及肿瘤旁组织MMP2及CD34的表达,采用IPP图像分析软件分析肿瘤组织、肿瘤旁组织MMP2及CD34的平均光密度。结果:（1）.各组均未见肝内、腹膜转移,无腹水形成。（2）.A组、B组、C组肿瘤最大径分别为（3.15±0.21）cm,（2.43±0.32）cm和（2.16±0.29） cm（P=0.000） ;肿瘤体积分别为（2.97±0.57） cm3 ,（2.00±0.68） cm3和（1.33±0.27）cm3（P=0.000） ;肿瘤重量分别为（4.39±0.40）g,（3.78±0.65）g和（3.36±0.41）g（P=0.000）。（3）. MMP2表达:A组、B组、C组肿瘤组织MMP2平均光密度分别为0.2052±0.0167,0.1980±0.0332,0.1711±0.0284（P=0.022）;肿瘤旁组织MMP2平均光密度分别为0.2081±0.0199,0.1726±0.0073,0.1396±0.0079 （P=0.000）。（4）. CD34表达:A组、B组、C组肿瘤组织CD34平均光密度分别为0.2310±0.0226 , 0.2219±0.0154 , 0.1886±0.0180（P=0.000）;肿瘤旁组织CD34平均光密度分别为0.2198±0.0505,0.1761±0.0237,0.1108±0.0046（P=0.000）。结论:（1）. CO₂气腹可促进兔肝VX2肿瘤生长;MMP2促进肿瘤血管新生;（2）. MMP抑制剂对CO₂气腹促进兔肝VX2肿瘤生长有一定抑制作用;（3）. MMP表达改变可能是腹腔镜手术后肿瘤生长转移的重要原因之一。更多还原

【Abstract】 Objective: To detect the expressions of MMP2 and CD34 proteins in rabbit liver VX2 tumor during CO₂ environment, so as to explore the effect of CO₂ on invasion, growth and metastasis in malignant tumor, and infer its mechanism from our findings.METHODS:Healthy male New Zealand rabbits weighing 2.5³.5kg were employed to establish models. After feeding one week under the same condition, thirty rabbits which were successfully implanted VX2 into liver were divided into 3 groups: Group A（Pneumoperitoneum group, n=10）, Group B（MMPi group, n=10）, Group C（Pneumoperitoneum-free group, n=10）.Group A and Group B received laparoscopic operation under condition that CO₂ pressure was 7.5mmHg, CO₂ flow was 0.4L/min. A Trocar was put into abdominal cavity at right lower quadrant. Five millilitres normal saline were administrated through intraperitoneal injection in Group A when the operation began, while 1,10-Phenanthroline （25mg/kg） were administrated through intraperitoneal injection in Group B. Pneumoperitoneum lasted for 3 hours. Group C did not establish pneumo- peritoneum instead of suspending rabbits’abdominal wall, the other treatments were the same as Group A. All animals were killed by euthanasia after raising 14 days under the same condition. We observed the tumor growth, measured tumor maximum diameters, tumor volume, tumor weight, and detected MMP2 and CD34 expressions both in tumor tissue and paratumor tissue by immunohistochemistry stain. Then,we analyzed the mean optical density of MMP2 and CD34 proteins by IPP system （Image-pro-plus 5.0.2）.RESULTS: （1）.No metastases or ascites were observed. （2）.Tumor maximum diameter was （3.15±0.21）cm in Group A,（2.43±0.32）cm in Group B and （2.16±0.29） cm in Group C（P=0.000）. Tumor volume was （2.97±0.57） cm3 in Group A,（2.00±0.68） cm3 in Group B,and （1.33±0.27） in Group C（P=0.000）. Tumor weight was （4.39±0.40）g in Group A,（3.78±0.65）g in Group B and （3.36±0.41）g in Group C（P=0.000）. （3）. Expressions of MMP2 protein: In Group A, B, C, respectively, the mean optical density was 0.2052±0.0167,0.1980±0.0332,0.1711±0.0284 （P=0.022） in tumor tissue and 0.2081±0.0199, 0.1726±0.0073, 0.1396±0.0079 （P=0.000） in paratumor tissue. （4）. Expressions of CD34 protein: In Group A, B, C, respectively, the mean optical density was 0.2310±0.0226, 0.2219±0.0154, 0.1886±0.0180 （P=0.000） in tumor tissue and 0.2198±0.0505, 0.1761±0.0237, 0.1108±0.0046 （P=0.000） in paratumor tissue. CONCLUSION: 1.CO₂ gas promotes the growth of VX2 tumor in rabbit liver. MMP2 stimulates tumor angiogenesis. 2. Matrix metallo- proteinase inhibitor can partially inhibit the growth of VX2 tumor in rabbit liver. 3. The change of expressions of MMP2 may play an important role in malignant tumor growth and metastasis after laparoscopic surgery.更多还原