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目的:通过观察华蟾素酯蟾毒配基类有效组分在体内对免疫相关细胞因子的影响,分析药物对机体免疫系统的调节作用,为阐明其免疫作用机制提供药理学和药效学依据。方法:体外培养人胰腺癌肿瘤细胞As PC-1,建立As PC-1胰腺癌荷瘤裸鼠动物模型,待给药结束后,通过流式细胞术的高通量多因子检测技术对小鼠血清进行粒细胞巨噬细胞集落刺激生物因子(GM-CSF),肿瘤坏死因子-α(TNF-α),干扰素-γ(IFN-γ),白细胞介素-1α(IL~(-1)α),白细胞介素-2(IL-2),白细胞介素-4(IL-4),白细胞介素-5(IL-5),白细胞介素-6(IL-6),白细胞介素-10(IL~(-1)0)检测,并用蛋白质免疫印迹法(Western blot)检测组织中相应细胞因子的蛋白含量,评价其体内免疫效果。结果:酯蟾毒配基类有效组分体内作用As PC-1胰腺癌荷瘤裸鼠免疫效果观察发现,与吉西他滨和华蟾素注射液比较,配基类组分能更多的增强细胞因子的表达。同时低剂量组对脾脏有一定的保护作用,高剂量组对脾脏的作用与华蟾素注射液和化疗药物吉西他滨相当。结论:华蟾素酯蟾毒配基类有效组分在体内能引起多种免疫相关细胞因子的分泌,增强机体的免疫功能,但具体的免疫作用机制还有待进一步研究。
OBJECTIVE: To investigate the effect of drugs on the immune system by analyzing the effects of the active components of the bufalin toxin on the immune-related cytokines in vivo and to provide pharmacological and pharmacodynamic evidence for elucidating the mechanism of immune action . METHODS: As PC-1 human pancreatic cancer cell line was cultured in vitro. Animal models of As PC-1 pancreatic cancer bearing nude mice were established. After the administration, high-throughput multi-factor flow cytometry Serum levels of GM-CSF, TNF-α, IFN-γ, IL-1α and IL- α, interleukin-2, interleukin-4, interleukin-5, interleukin-6, interleukin- -10 (IL ~ (-1) 0). Western blot was used to detect the protein content of the corresponding cytokines in the tissue and the in vivo immune response was evaluated. Results: The immune effect of Aspergillus fumigatus active ingredients in vivo on As PC-1 pancreatic cancer-bearing nude mice was observed. Compared with gemcitabine and cinobufacini injection, ligand-like components could enhance cytokines expression. At the same time, the low dose group had a certain protective effect on the spleen. The effect of the high dose group on the spleen was comparable to that of cinobufacini injection and chemotherapy drug gemcitabine. CONCLUSION: The active components of cinobufogenin toxoid can induce the secretion of multiple immune-related cytokines and enhance the immune function in vivo. However, the specific immunological mechanism remains to be further studied.