摘要
目的:观察具有诱导癌细胞凋亡的作用,并作为免疫调节剂的温莪术提取物β-榄香烯是否可协同抗CD3-抗胶质瘤双特异性抗体免疫抑制胶质瘤生长。方法:实验于2003-12/2005-08在苏州大学附属第二医院脑肿瘤研究室完成。①选用重度联合免疫缺陷SCID小鼠30只,雌雄不拘,鼠龄4～6周。温莪术提取物β-榄香烯由大连市医药科学研究所馈赠。采用化学偶联法将由解放军第二军医大学长海医院免疫室提供的CD3单抗和本室制备的SZ39抗人脑胶质瘤单抗制备为双特异性抗体。②取培养生长的人脑胶瘤体外细胞系SHG-44细胞3×108L-1,于人外周血单个核细胞转输后次日,直接注射于SCID鼠右腋皮下。将种瘤后SCID小鼠按随机抽签法分为5组:β-榄香烯+抗体+白细胞介素2+单个核细胞组:每天腹腔注射β-榄香烯(1.5mg/只)和双特异性抗体200μg/只,每天尾静脉注射白细胞介素2(5×105U/只),接种肿瘤前1天及第14天尾静脉注射人外周血单个核细胞(3.16×1010L-1,0.5mL,取自苏州大学附属第二医院健康志愿者2名外周血);抗体+白细胞介素2+单个核细胞组:注射3种物质剂量和方法同上组;白细胞介素2+单个核细胞组:注射2种物质剂量和方法同上组;外周单个核细胞组:注射外周单个核细胞剂量和方法同上组;对照组:尾静脉注射培养液。③记录肿瘤增殖的潜伏期,即接种日至皮下扪及微结节的时间。接种肿瘤28d后,称肿瘤质量。荧光原位杂交法测算每克肿瘤人淋巴细胞数量。苏木精-伊红染色,Olympus光学显微镜(×200)下观察肿瘤组织病理学变化。结果:SCID小鼠30只均进入结果分析。①神经胶质瘤组织病理学观察结果:β-榄香烯+抗体+白细胞介素2+外周血单个核细胞组和抗体+白细胞介素2+外周单个核细胞组肿瘤细胞有部分坏死,并有淋巴细胞浸润,其余各组形态相近,淋巴细胞浸润相对较少。②肿瘤增殖潜伏期:β-榄香烯+抗体+白细胞介素2+单个核细胞组明显长于其他4组(P<0.05),抗体+白细胞介素2+单个核细胞组明显长于白细胞介素2+单个核细胞组、外周单个核细胞组和对照组(P<0.05),白细胞介素2+单个核细胞组明显长于外周单个核细胞组和对照组(P<0.05)。③肿瘤质量:β-榄香烯+抗体组明显高于其他4组(P<0.01)。④肿瘤中人淋巴细胞浸润数量:β-榄香烯+抗体组明显多于其他4组(P<0.01)。结论:温莪术提取物可协同双特异性抗体抑制神经胶质瘤生长。
AIM： To observe the inhibitory effects of curcuma zedoary extract β- elemene, which can induce the tumor cell apoptosis and be used as an immunomedulator, cooperating with anti-CD3/anti-glioma bispecific antibodies （BsAb） on glioma.
METHODS： The experiment was completed in the Institute of Tumor, the Second Affiliated Hospital of Suzhou University from December 2003 to August 2005. ①A total of 30 severe combined immunedeficiency （SCID） mice of either sex and 4-6 weeks old were selected. Curcuma zedoary extract β-elemene was presented by Dalian Institute of Medical Sciences. Chemical coupling method was applied to prepare BsAb with CD3 monoantibody （provided by Changhai Hospital of the Second Military Medical University of Chinese PLA） and. SZ39 anti-human brain glioma antibody （prepared by the Second Affiliated Hospital of Suzhou University）. ②3×10s L^-1 SHG-44, the cultured human brain glioma in vitro cell line, were injected directly into right axilla of mice on the second day after human peripheral blood lymphocyte （HuPBL） transportation. Then these SCID mice were randomized into 5 groups： A group [β-elemene ＋BsAb ＋ interleukin-2 （IL-2）＋HuPBL]： The intraperitoneal injection of 1.5 mg β- elemene and 200μg BsAb was given to each mice everyday. In addition, 5×10^5 U IL-2 was daily injected intravenously into each mice tail, and HuPBL （3.16×10^10 L^-1, 0.5 mL, obtained from the peripheral blood of two healthy volunteers of the Second Affiliated Hospital of Suzhou University） was injected intravenously into each mice tail on one day before and 14 days after tumor inoculation; B group （BsAb＋IL-2＋HuPBL）; C group （IL-2＋ HuPBL）; D group （HuPBL）; Control group： Culture medium was injected into tail veins, while other groups were similar with A group in dosage and administration methed.③The latent period of tumor growth was estimated by the time from the inoculation day to subcutaneous nodules palpation. The tumor was weighted at the 28^th day after inoculation, and the number of lymphocytes in each gram tumor was calculated with fluorescence in situ hybridization （FISH）; Then histopathological change of tumor were observed by hematoxylin-eosin stain and Olympus light microscope （×200）.
RESULTS： All the 30 mice were involved in the result analysis.①The histopathology of neuroglioma： A group and B group occurred necrosis partly and lymphocytes infiltration, while there was similar form and few lymphocytes infiltration in other groups. ②The latent period of tumor was obviously longer in A group than in other 4 groups （P 〈 0.05）, and obviously longer in B group than in c group, D group and control group （P 〈 0.05）, also obviously longer in C group than D group and control group （P 〈 0.05）.③The weight of tumor： It was notably higher in A group then in other 4 groups （P 〈 0.01）. ④The number of lymphocytes infiltration in A group was siguificantly increased compared with other 4 groups （P 〈 0.01）.
CONCLUSION： β-elemene can effectively inhibit the growth of glioma cooperating with BsAb.
出处
《中国临床康复》
CSCD
北大核心
2006年第23期124-126,F0003,共4页
Chinese Journal of Clinical Rehabilitation
