双歧杆菌联合顺铂对黑色素瘤小鼠的治疗作用及机制

Inhibitory activity of Bifieobacterium adolescent and cisplatin on the growth of melanoma in mice and its regulatory mechenism

目的:探讨双歧杆菌、顺铂及二者联合对黑色素瘤小鼠的抑瘤作用及其机制.方法:建立黑色素瘤荷瘤小鼠模型,分别以皮下及腹腔注射双歧杆菌和顺铂,处死小鼠后测瘤质量计算抑瘤率,病理切片后HE染色观察瘤组织及瘤细胞变化,流式细胞仪测瘤细胞周期,MTT法测小鼠淋巴细胞活性,CD3,CD4,CD8双荧光标记法测小鼠T细胞亚群变化.结果:注射1×1010cfu/L的双歧杆菌其抑瘤率可达54.0%,与顺铂组相似,与顺铂联合作用后其抑瘤作用更强,达74.4%;病理切片显示双歧杆菌抑制瘤细胞生长,且加强了顺铂杀伤瘤细胞的作用;流式细胞仪检测结果显示:双歧杆菌作用后瘤细胞生长阻滞于G1期,顺铂作用后瘤细胞生长阻滞于S期;双歧杆菌作用后小鼠细胞免疫功能增强,主要表现为CD4+比例增加,与正常组比较差异有统计学意义(P<0.01).结论:双歧杆菌联合顺铂对黑色素瘤小鼠具有较强的抑瘤作用,其机制与抑制瘤细胞生长及增强小鼠细胞免疫功能有关.

AIM： To study the inhibitory activity of Bifidobacte- rium adolescent and cisplatin on the growth of melanoma （B16） in mice and its regulatory mechanism. METHODS： B16 cells were injected to C57/65 mice to establish animal model bearing melanoma. Bifidobacterium adolescent and cisplatin were injected subcutaneously and intraperitoneally respectively. The rates of in- hibitory activity on the growth of melanoma （ B16 ） were calculat- ed. Pathological changes of the tumor, tissue and cells were observed by HE staining. B16 cell cycles were determined by flow cytometry. Lymphocytes proliferation was measured by MTT colo- rimetric assay. T-cell subsets were monitored by double labelled fluorescence. RESULTS： The rate of inhibitory activity on the growth of melanoma （B16） by injecting bifidobacterium of 1 x 10^10 cfu/L reached 54%, similar with that in cisplatin group. The inhibitory activity was most strong after bifidobacterium was com- bined with cisplatin, with the inhibition rate reached 74.45%. Bifidobacterium treatment inhibited proliferation of B16 cells and enhanced the killing activity of cisplatin on B16 cells. Flow cytometry revealed that the B16 cells proliferation was inhibited to G1 stages after bifidobacterium treatment, and to S stages after cisplatin treatment. T-cell immune activity was enhanced after bifidobacterium treatment, which was featured by the increased CD4^＋ ratio, and the difference was significant compared with the normal group （ P 〈 0. 01 ）. CONCLUSION： Combination of bifidobacterium with cisplatin exerts strong inhibitory activity on the growth of melanoma. The mechanism is correlated to the inhibitory activity on B16 cell proliferation and the enhancedimmune activity in mice.