摘要
目的探讨腹水传代与体外培养walker256细胞接种建立SD大鼠骨癌痛模型的可行性并验证其可靠性。方法体外培养细胞与大鼠腹水传代培养Walker256细胞，种植于SD大鼠左胫骨上端。大鼠分为Hank’s液对照组（N组）、热杀死肿瘤细胞组（K组）、体外培养的肿瘤细胞种植组（V组）和腹水肿瘤细胞种植组（A组），每组8只。通过疼痛行为学、镇痛药效学、影像学、病理组织学等检查评估肿瘤及疼痛发生发展情况。结果大鼠接种后肛温无明显变化。A组与V组大鼠术后第6天开始左侧后肢活动度开始下降，X线摄片可见左胫骨上端骨小梁小缺损；第12天X线摄片示骨皮质有破坏，放射性核素显像可见接种区反应性骨形成活跃；第14天增重开始减缓；第18天X线摄片显示大片骨质缺损，软组织肿块形成，左胫骨病理切片示骨癌细胞生长。模型大鼠接种后第6～18天，机械性压爪缩爪阈值、触痛觉超敏von Frey阈值、左侧后肢负重进行性下降（P〈0．01）。术后第15天注射吗啡后，模型大鼠机械性压爪缩爪阈值呈剂量依赖性增高，而纳洛酮可拮抗此效应。结论经腹水传代与体外培养Wal ker256细胞均可用于SD大鼠骨癌痛模型的建立；经腹水传代建模更为简便。
Objective To investigate the possibility of estabhshing rat model of bone cancer pain using cancer cells cultured in vitro or by ascites passaging and verify the reliability of this method. Methods Syngeneic SD rat carcinoma cells of the line Walker 256 were cultured in vitro and inoculated into the peritoneal cavity of SD rats respectively. Two kinds of Walker 256 cell suspension were made. Thirty-two SD rats were randomly divided into 4 equal groups： Group N （undergoing injection of Hank＇s solution into cavitas medullaris of left tibia）, Group K （undergoing injection of heat-killed Walker 256 cells）, Group V （injected with Walker 256 cells cultured in vitro）, and Group A （injected with Walker 256 cells passaged in ascites）. After 6, 12, and 18 days, the rats underwent rcentgenography. Radio nuclide emission computed tomography （ECT） was conducted 12 days later. And MRI was conducted 15 days later. Thermal withdrawal latency （TWL） and pressure withdrawal threshold （PWT） were measured. Von Frey threshold and weight bearing of left hind limb were examined. On the 18th day after the estabhshment of model the rats were killed with their left tibia taken out to undergo microscopy. Results The rats of Groups A and V began to display decrease of left hint limb activity and roentgenography showed minute defect of bone trabecula in the proximal epiphysis by day 6. By day 12 reentgenography showed multiple defect of bone trabecula, ECT showed reactive bone formation. The rats of Groups A and V displayed signs of weight loss by day 14, and by day 18 roentgenography showed full thickness bicortical bone loss and formation of soft tissue tumor. Histological examination 18 days later revealed that the bones inoculated with live cells showed infiltration of bone marrow spaces by malignant tumor. The PWT values gradually decreased since day 6 to day 18, and the PWT values in this period of Groups A and V were all significantly lower than those of Groups K and N （all P〈0.01）.The von Frey values gradually decreased since day 6 to day 18,and the von Frey values in this period of Groups A and V were all significantly lower than those of Groups K and N （ all P 〈 0.01 ）. The weight bearing value of left hind limb gradually decreased and the weight beating difference between the 2 hind limbs gradually increased since day 6 to day 18, and the values of weight bearing difference between the 2 hind limbs of Groups A and V were significantly higher than those of Groups K and N （ all P 〈 0.01 ）. Sixteen rats underwent subcutaneous injection of morphine, and the PWT values increased 20, 30, and 40 min later in a dose-dependent manner （ P 〈 O. O1 ）. Naloxone injected 1 h later antagonized the analgesic effect of morphine. Conclusion A SD rat model of bone cancer pain has been successfully established by using syngeneic rat bone carcinoma cells cultured in vitro or in vivo, and the latter being more convenient.
出处
《中华医学杂志》
CAS
CSCD
北大核心
2008年第13期880-884,共5页
National Medical Journal of China
关键词
模型
动物
大鼠
腹水
接种
疼痛
胫骨
Models,animal
Rats
Ascites
Vaccination
Pain
Tibia
