修回日期: 2002-01-20
接受日期: 2002-02-07
在线出版日期: 2003-09-15
观察5-FU白芨微球经肝动脉注射对兔VX2移植性肝癌的治疗效果.
50只兔VX2移植性肝癌模型, 随机分为5组 (n = 10), 采用直视下直接穿刺血管的方法, 分别经肝动脉注入生理盐水1 mL; 5-FU溶液1 mL; 超液态碘油0.3-0.4 mL加5-FU溶液1 mL; 100 g/L白芨胶0.4 mL加5-FU溶液1 mL; 5-FU白芨微球10 mg/kg后, 观察肿瘤生长情况、坏死程度及肝功能变化等.
5-FU白芨微球栓塞后肿瘤生长受到显著抑制, 肿瘤生长率低于对照组(P < 0.01)及碘油组(P < 0.05). 肿瘤坏死以重度为主, 有2例完全坏死, 优于碘油组及白芨胶组. 但对正常肝组织损伤亦相对较重.
5-FU白芨微球具有良好的血管栓塞作用, 使用方便、安全, 是一种理想的末梢性栓塞剂.
引文著录: 李欣, 冯敢生, 郑传胜, 柳曦, 孔健. 经肝动脉注射5-FU白芨微球治疗兔VX2移植性肝癌. 世界华人消化杂志 2003; 11(9): 1337-1340
Revised: January 20, 2002
Accepted: February 7, 2002
Published online: September 15, 2003
To observe the therapeutic efficacy of 5-FU bletilla striata microspheres inFUsed through hepatic artery against rabbit VX2 transplanted hepatoma.
Fifty rabbits bearing VX2 transplanted hepatoma were randomly divided into five groups (n =10). Normal saline 1 mL, 5-Fu solutions 1 mL, ultr-liquor-lipiodol 0.3-0.4 mL and 5-Fu solutions 1 mL, 100 g/L Bletillaolloid 0.4 mL and 5-Fu solutions 1 mL, 5 FU-bletilla striate microspheres 10 mg/kg were infused though hepatic artery respectively. The tumor growth rates, necrosis rates and hepatic function were compared among the 5 groups.
5-FU bletilla striata microspheres had a very good vessel embolization function. In 5FU-bletilla striata microspheres group, the tumor was significantly inhibited, tumor growth rate was lower than tant in the control group (P <0.01) and the lipiodol group (P <0.05). Tumor necrosis grades were also more higher compared with to the other 4 groups. Complete necrosis was found in 2 of 10 rabbits in 5FU-bletilla striata microspheres group, which was more severe than that in the lipiodol group and bletillaolloid group. But the damage of normal liver tissues was also more serious.
5-FU bletilla striata microsphere is a safe and effective peripheral embolization agent.
- Citation: Li X, Feng GS, Zhen CS, Liu X, Kong J. Therapeutic efficacy of 5-FU bletilla striata microspheres infused through hepatic artery against rabbit VX2 transplanted hepatoma. Shijie Huaren Xiaohua Zazhi 2003; 11(9): 1337-1340
- URL: https://www.wjgnet.com/1009-3079/full/v11/i9/1337.htm
- DOI: https://dx.doi.org/10.11569/wcjd.v11.i9.1337
介入化疗栓塞术是治疗肝癌最有效的方法之一 [1-16], 而栓塞剂及其剂型的选择是化疗栓塞治疗的研究重点 [17-25]. 白芨是一种理想的栓塞剂 [26-32]. 我们制备了5-FU白芨微球, 并用其对兔VX2移植性肝癌模型进行栓塞化疗研究, 以期阐明该微球对肝癌的疗效及作用机制, 为其在临床的合理应用提供可靠的理论依据.
♂新西兰大白兔50只, 重量2-2.5 kg, 由华中科技大学同济医学院实验动物中心提供; VX2肿瘤细胞株由美国ATCC公司引进; 超液态碘油为法国Aulnay-sous-Bios公司产品, 5-FU注射液为上海旭东海普药业有限公司产品. 白芨胶, 5-FU白芨微球为我院药剂科制备, 制备方法及质量监控见文献 [30,31].
兔VX2移植性肝癌模型的建立参照Prat (Hepatology 1995; 21: 832)的方法并加以改进[33], 将瘤块接种于肝左外叶. 50只荷瘤兔, 随机分为5组, 每组10只, 即对照组: 经肝动脉灌注生理盐水1 mL; 5-FU组: 经肝动脉灌注5-FU溶液1 mL; 碘油+5-FU组: 经肝动脉灌注超液态碘油0.3-0.4 mL加5-FU溶液1 mL; 白芨胶+5-FU组: 经肝动脉灌注100 g/L白芨胶0.4 mL加5-FU溶液1 mL; 白芨微球组: 经肝动脉灌注5-FU白芨微球10 mg/kg. 于瘤块接种后14 d, 再次开腹, 将胃向左外完全翻出, 充分暴露肝门区后, 用自制27 G穿刺针直接穿刺肝固有或肝左动脉, 用动脉夹固定, 并暂时阻断肝右动脉血流. 造影及分别注射各组药物后, 对穿刺点局部加压止血, 逐层缝合、消毒, 动物分笼饲养. 术后1, 2, 3 wk进行CT及MR扫描, 依扫描所得瘤灶最大层面测定肿瘤的体积, 根据公式计算肿瘤生长率(growth rate, GR), GR=V治疗前/V治疗后×100 %. 术后2, 3 wk末各处死5只动物, 取肝组织, 以40 g/ L多聚甲醛固定24 h, 石蜡包埋, 每份标本分2-3 处取材, HE染色, 切片. 光镜下观察肿瘤组织坏死范围, 分为轻度(0-30%)、中度(30-70 %)、重度(70-100 %). 于术前、术后3, 7, 14, 21 d, 经耳缘静脉取血, 血样进行肝功能检查(TB、ALT). 由全自动血生化分析仪完成.
统计学处理 采用SPSS for Windows 统计软件, 对成组资料进行方差分析、组间两两比较采用Newman-keuls检验, 对计数资料作x2 检验, 均以P <0.05为有统计学意义.
5-FU白芨微球组, 栓塞后大体标本显示肝左叶可见斑片、片状局灶梗死灶, 呈苍白色, 界限清楚, 肿瘤切面可见中心明显坏死(图1). 光镜检查可见肿瘤明显凝固性坏死, 但大多数病例仍可见残余病灶, 及不同程度的出血、胆汁郁积. 瘤周可见较厚的纤维间隔, 其内可见较多炎性细胞浸润(图2). 正常肝组织亦见不同程度变性坏死, 其程度较碘油、白芨胶组栓塞组重, 肝内未见小脓肿形成. 5-FU白芨微球镜下呈半透明状, 多栓塞于小叶间动脉、微小动脉和毛细血管前微动脉水平, 栓塞血管彻底(图3). 变性、坏死区内的血管中微球多见. 被栓塞血管可出现(血管)炎性改变, 如血管壁增生、水肿, 局部有少量炎性细胞浸润, 常见继发性血栓形成, 并可见管腔闭塞. 肝窦、毛细血管、小静脉内及肺部标本未见栓塞. 但在栓塞区域门脉小分支内常可见血栓形成.
治疗前肿瘤体积差异均无显著性, 治疗后对照组兔肿瘤体积增大最明显, 5-FU白芨微球组兔肿瘤体积小于其余各组, 并有4例肿瘤体积缩小(表1). 对照组及5-FU组的肿瘤坏死程度以轻度坏死为主, 碘油组以中度为主, 兼有部分重度坏死, 白芨胶及白芨微球组则以重度坏死为主. 5-FU白芨微球组10例中, 8例见肿瘤重度明显坏死, 其中2例完全坏死, 未见存活的癌细胞, 显著大于碘油栓塞组(P <0.05). (表2)栓塞组术后ALT升高, 3 d达峰值, 然后逐渐下降, 2 wk左右降至正常, 总胆红素水平变化不明显.
分组 | 肿瘤体积(cm3) | 术后3 wk肿瘤增长率 (倍) | |||
栓塞前 | 栓后1 wk | 栓后2 wk | 栓后3 wk | ||
对照组 | 0.96±0.26 | 5.24±1.55 | 13.1±2.96 | 26.2±5.31 | 28±5.82 |
5-FU化疗组 | 0.99±0.49 | 5.22±1.96 | 11.9±2.16 | 23.1±4.76 | 25.72±8.28 |
碘油+5-FU组 | 0.99±0.39 | 1.95±0.27a | 3.00±1.18a | 6.66±1.98a | 6.99±1.94a |
白芨胶+5-FU组 | 1.00±0.31 | 1.43±0.29a | 2.14±0.88a | 4.20±1.76a | 4.27±0.99a |
5-FU白芨微球组 | 1.00±0.25 | 1.17±0.26ac | 1.72±1.24a | 2.84±1.69ac | 2.97±1.86ac |
中药白芨是一种集载体、栓塞、缓释及抗肿瘤作用为一体的较理想的栓塞剂[26-29], 在白芨胶的基础上, 我们进一步制备了5-FU白芨微球, 并用其对兔VX2移植性肝癌模型进行栓塞化疗研究, 以判断其疗效及机制. 兔VX2肿瘤是一种常用的富血管性肿瘤 [34-36], 我们在国内外首次采用了直视下肝动脉直接穿刺法, 此法实验成功率高, 并可在栓塞的同时阻断肝右动脉, 达到了选择性栓塞的目的, 防止了误栓, 提高了术后动物的存活率. 结果显示, 5-FU白芨微球在抑制肿瘤生长率方面显示了较好的疗效, 使肿瘤坏死更彻底, 有4例肿瘤缩小, 并有2例镜下可见肿瘤完全性坏死, 优于单纯化疗组及碘油栓塞组, 与白芨胶栓塞组相近并稍优. 但同时对正常肝脏的损伤也相对较重, 大体肝标本可见斑片, 片状坏死区, 镜下可见正常肝组织变性严重, 并见点状、灶状坏死. 术后ALT有一过性升高, 但可较快恢复.
5-FU白芨微球有良好的末梢栓塞作用, 5-FU白芨微球粒径为50-200 mm, 在栓塞后病理切片上显示其栓塞水平为肝动脉3级以下分支, 最远可达肝窦前动脉水平, 其层面在动-门脉间及肝内潜在吻合支水平以远 [27,28], 减少了栓塞后侧支循环形成的可能性. 栓塞区域门脉末梢分支内常可见血栓形成, 推测微球本身及其分解产物可能通过动-门脉间吻合支进入门脉末梢 [28,29], 起到了动-门脉双重栓塞的作用. 白芨有较强的促凝血作用, 5-FU有较强的致血管炎作用, 可刺激血管产生血管炎、继发性栓塞, 加重了栓塞程度, 延长了栓塞时间. 白芨微球具有较好的载体和缓释作用, 可维持5-FU瘤内局部高浓度, 延长药物在肿瘤内的作用时间, 与栓塞起协同增效作用. 白芨本身有一定的抗肿瘤作用, 其主要成分黏液质是一种广谱抗肿瘤成分, 对肿瘤的发生、发展均有显著的抑制作用.
与造影剂混合后, 微球悬浮性和分散性好, 不粘结, 易经小管径导管注射, 易于操作, 使用方便. 而肝窦、毛细血管、小静脉内及肺部标本中未见微球栓塞作为栓塞剂, 说明5-FU白芨微球注射后能固定于靶器官内, 没有滤过毛细血管, 造成异位栓塞. 所有动物于栓塞后均生长良好. 这些结果均说明5-FU白芨微球具有良好的生物相容性, 安全、可靠. 本研究表明, 5-FU白芨微球具有良好的血管栓塞作用, 使用安全、方便, 是一种理想的末梢性栓塞剂, 但对正常肝组织的损伤也相对较重, 故在实际应用中应注意选择适当病例, 采用超选择性插管, 尽量避免损伤正常肝组织.
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