论文标题:腹腔镜气腹对大鼠肝癌模型肿瘤体积、细胞凋亡及增殖影响的研究
The Study of CO_2 as Gas Source in Laparoscopy with Tumor Volume, Apoptosis and Proliferation in HCC Rat Model
论文作者 晏益核
论文导师 卢榜裕,论文学位 硕士,论文专业 外科学
论文单位 广西医科大学,点击次数 75,论文页数 39页File Size3548k
2005-05-01论文网 http://www.lw23.com/lunwen_90778147/ 腹腔镜; 气腹;大鼠;肝癌; 肿瘤体积;凋亡;增殖
Laparoscopy ; Pneumoperitoneum ;Hepatic carcinomar; rat Tumor volume ; Apoptosis; Proliferation
目的:利用建立的移植性大鼠肝癌模型,评估及比较不同种类的膨腹气体原对大鼠肝癌模型肿瘤体积、细胞凋亡及增殖的影响,以此探讨气腹对肿瘤的生长和转移的影响。方法:1)建立移植性大鼠肝癌模型:将细胞株Walker256注射到雄性Wistar 大鼠(60-80g)的四肢皮下,6 天后将皮下出现的肿块取出,再将肿块(体积约1×1×1mm3)种植到大鼠的肝脏内。2)手术干预:模型建立后10 天,将总共100 只大鼠(体重200-250g)随机分为5 组:二氧化碳组(CO2)、氦气组(He)、氮气组(N2)、开腹组及单纯麻醉组,每组各20只。气腹组用穿刺管将不同气体输入腹腔,压力8mmHg,持续1 小时;开腹组在腹壁正中从剑突下至耻骨联合上做一切口,暴露腹腔,持续1 小时后缝合;单纯麻醉组只插入穿刺管,持续1 小时。3)检测肿瘤体积、细胞凋亡及增殖:在术后1 天处死各组存活鼠的一半,另一半于术后7 天处死;取出肿瘤,测量体积,之后制成切片,分别采用TUNEL 法检测肿瘤细胞凋亡,免疫组织化学方法标记cyclin-PCNA 来检测增殖指数。结果:术后1 天和7 天,CO2 组、He 组、N2组、开腹组及单纯麻醉组肿瘤体积比较无统计学显著差异(Kruskal-Wallis test: χ2=2.642 P=0.619;χ2=0.209 P=0.995);增殖指数比较也无统计学显著差异(ANOVA:F = 0.554, P=0.697;F=1.236, P=0.323);术后1天,肿瘤细胞凋亡比较无统计学显著差异(ANOVA:F=0.413, P=0.798);而术后7 天,凋亡比较有统计学显著差异(F=5.698, P=0.002)。结论:CO2气腹并没有促进大鼠肝癌模型肿瘤的生长及改变肿瘤细胞的生物学特征;He 气腹有诱导肿瘤凋亡的作用。
Objective: In this study, we evaluated and compared effect of differentgas sources used in the laparoscopy with tumor growth, apoptosis andproliferation of tumor cells in HCC model. Methods: 1) A transplantedhepatic carcinoma rat model was set up. The Walker256 cell linesuspensions were subcutaneously injected into the limb of Wistar malerat (weight 60-80g). The mass (approximately 1 ×1 ×1 mm3) wastransplanted into the rat liver on the day 6 after the mass appeared.2) The operation in the rats was performed. On the day 10 after modelwas established, total 100 HCC rats (weight 200-250 g) were randomizedinto the carbon dioxide, helium, nitrogen, laparotomy, and anesthesiacontrol groups. The insuflation groups underwent CO2, He, and N2pneumoperitoneum(8mmHg), respectively, for 60 min via an angiocatheter.The laparotomy group only underwent a midline incision from xiphoidto pubis and closed 60 min later. The anesthesia control group was justinserted with an angiocatheter. 3) Tumor volume and apoptosis as wellas proliferation of tumor cells were determined. Tumors were excisedfrom half the survival rats in each group on the day 1 and 7postoperation respectively. The tumor volumes were measured, and thensections of the tumor tissues were made for detecting the apoptosiswith TUNEL and proliferation by cyclin-PCNA index withImmunohistochemistry. Result: Either on the day 1 or 7 postoperation ,there was no statistical significant difference between the groups inthe tumor volume (Kruskal-Wallis test: χ2=2.642 P=0.619;χ2=0.209P=0.995). Also no significant difference was found in proliferativeindex (ANOVA: F=0.554 P=0.697;F=1.236 P=0.323) and in apoptotic rateon the day 1 postoperation(ANOVA: F=0.413 P=0.798). However, there wassignificant difference in apoptotic rate on the day 7 postoperation(F=5.698 P=0.002). Conclusion: Our data demonstrated that CO2 as gassource in pneumoperitoneum did not play role in enhancing the tumorgrowth and affecting the biological features of tumor in this HCC ratmodel; He as gas source can induce the apoptosis of tumor in this model.
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