亚高温盆腔区域热疗对原位膀胱肿瘤大鼠免疫状态影响的研究

The immunologic changes of rat loading with orthotopic bladder tumor post treatment by non-invasive pelvic cavity regional moderate hyperthermia

目的研究亚高温盆腔区域热疗致原位膀胱肿瘤大鼠发生热休克反应后机体免疫状态的改变。方法经尿道膀胱灌注N-甲基亚硝基脲(MNU)制备大鼠膀胱肿瘤动物模型35只,随机分入短期组,包括肿瘤对照组(A组)、单次热疗组(B组)、间隔24h再次热疗组(C组)、间隔72h再次热疗组(D组)和长期组,包括肿瘤对照组(E组)、间隔24h再次热疗组(F组)、间隔72h再次热疗组(G组)。用内生场热疗系统从体外加热大鼠盆腔至41℃。热疗后对肿瘤、引流淋巴结、脾脏中免疫细胞的变化进行病理研究,检测血清鼠IFN-γ含量。结果透射电镜观察到B、C、D组肿瘤组织中有较多中性粒细胞、巨噬细胞;F、G组中性粒细胞聚集,巨噬细胞吞噬凋亡小体。短期组中B、C、D组膀胱肿瘤组织、淋巴结及脾脏中S-100蛋白阳性细胞率均高于A组(P<0.01)。膀胱肿瘤组织及脾脏中CD8阳性细胞率、血清鼠IFN-γ含量A、B、C、D组间差异均无统计学意义(P=0.657、0.440、0.523)。长期组中F、G组的各项数据均高于E组(P<0.01)。结论随着亚高温盆腔区域热疗次数的增加肿瘤鼠的免疫细胞重新分布,有利于肿瘤抗原提呈及效应细胞攻击肿瘤。

Objective To study the immunologic changes of rats with orthotopic bladder tumor post treatment by non-invasive regional moderate hyperthermia in pelvic cavity. Methods 35 six-week old female inbred line F344 rats were instilled MNU transurethrally once every other week for 6 times. Groups： Short-term group contained 4 groups. Group A was control. Group B wag heated once. Group C w^ls heated twice with a interval of 24 h. Group D was heated twice inter- val 72 h. Long-term groups contained 3 groups. Group E was control. Group F was heated twice with a interval of 24 h. Group G heated twice with a interval of 72 h. Each group contained 5 rats. A rat pelvic was heated to 41 ~C and persisted an hour by NRL-00II Double RF Tumor Hyperthermia System. The pathologic changes of tumor, lymphonode of draining domain around bladder and spleen were detected by electron microscopy and light microscope respectively. The quantity of Rat IFN-y was detected by ELISA. Results Electron microscopy indicated that there were some neu- trophile granulocytes and macrophages emerged in tumor o{ group B, C, and D. The aggregation of neutrophile granulocytes and macrophages containing apoptosis bodies were observed in group F and G. The positive cells of S-100 protein in tumor, lymphonode and spleen of group B, C, and D were significant than that of group A （P d0.01）. The positive ceils of CD8 in tumor and in spleen and Rat IFN-T quanti- ties were non-significant difference among of group A, B, C and D respectively （P= 0. 657, 0. 440, 0. 523）. The positive cells of S-100 protein and CD8 in tumor, lymphonode of draining domain around bladder and spleen and Rat IFN-γ quantities in group F and G were significant than that of group E （P〈0.01）. Conclusions The redistribution of immunocytes of rats loading with orthotopic bladder tumor was followed with the increasing of frequency of non-invasive pelvic cavity regional moderate hyperthermia. These changes were benefit to tumor antigen presentation and ef- fective immunocytes attack to tumor cell.