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目的探讨双歧杆菌脂磷壁酸(lipoteichoi cacid of bifidobacterium,BLTA)对黑色素瘤B16小鼠Fas/FasL表达的影响。方法将黑色素瘤B16细胞接种于C57BL/6小鼠(n=40)皮下,待触及肿块后于荷瘤小鼠瘤周注射不同剂量的BLTA。实验动物分为4组,每组10只:生理盐水(对照)组、低浓度BLTA(50mg/LBLTA)组、中浓度BLTA(100mg/LBLTA)组、高浓度BLTA(150mg/LBLTA)组。采用MTT法检测荷瘤小鼠CTL杀伤活性;免疫组织化学染色检测肿瘤组织浸润的CD4+、CD8+淋巴细胞的表达情况;用RT-PCR检测肿瘤组织和脾脏组织Fas、FasL mRNA的表达变化;免疫组织化学染色和Western blot检测肿瘤组织和脾脏淋巴细胞Fas、FasL蛋白的表达变化。结果与对照组比较,BLTA处理各组荷瘤小鼠的CTL杀伤率明显增加(P<0.05),肿瘤组织内Fas、FasL mRNA和蛋白表达明显增加(P<0.05),而荷瘤小鼠脾脏组织的Fas、FasLmRNA和蛋白表达明显下调(P<0.05),肿瘤组织内CD8+淋巴细胞的浸润增多(P<0.05),并具有剂量依赖性。CD4+淋巴细胞的浸润无明显增加(P>0.05)。结论 BLTA能够通过调节Fas/FasL系统,增强CTL细胞的杀伤活性,促进肿瘤细胞的凋亡,逆转荷瘤小鼠的肿瘤免疫逃逸现象,进而提高机体的抗肿瘤作用。
Objective To investigate the effect of lipoteichoic acid lipoteichoic acid (BLTA) on Fas / FasL expression in melanoma B16 mice. Methods Melanoma B16 cells were inoculated subcutaneously in C57BL / 6 mice (n = 40). After tumor mass was reached, different doses of BLTA were injected peritumically into tumor bearing mice. The experimental animals were divided into 4 groups with 10 rats in each group: saline (control) group, low concentration of BLTA (50mg / LBLTA) group, medium concentration of BLTA (100mg / LBLTA) group and high concentration of BLTA (150mg / LBLTA) group. The cytotoxic activity of CTL in tumor-bearing mice was detected by MTT assay. The expression of CD4 + and CD8 + lymphocytes infiltrating tumor tissue was detected by immunohistochemical staining. The expressions of Fas and FasL mRNA in tumor and spleen were detected by RT-PCR. The expressions of Fas and FasL protein in tumor and spleen lymphocytes were detected by chemical staining and Western blot. Results Compared with the control group, the killing rate of CTL in BLTA-treated mice was significantly increased (P <0.05), while the expression of Fas and FasL mRNA and protein in tumor tissues was significantly increased (P <0.05) The expressions of Fas, FasL mRNA and protein were significantly downregulated (P <0.05), and the infiltration of CD8 + lymphocytes in tumor tissues was increased (P <0.05) in a dose - dependent manner. No significant increase in infiltration of CD4 + lymphocytes (P> 0.05). Conclusion BLTA can enhance the anti-tumor activity of CTL cells by regulating the Fas / FasL system, enhancing the cytotoxicity of CTL cells, promoting the apoptosis of tumor cells and reversing the tumor immune escape phenomenon in tumor-bearing mice.