白色念珠菌细胞壁不溶性β-葡聚糖（β-glucan）诱导外周血单核细胞TNF-α的表达

发布时间：2018-06-26 22:21

本文选题：白色念珠菌不溶性β-葡聚糖 + PBMC　；参考：《昆明医学院》2008年硕士论文

【摘要】： 【目的】白色念珠菌细胞壁不溶性β-葡聚糖(CAIBG)是一类生物反应调节剂(BRM),具有升白细胞、抗肿瘤、抗病毒等广泛的生物学活性,本研究前期实验已经成功地从白色念珠菌细胞壁中分离出CAIBG,并通过动物实验证实了CAIBG能对抗环磷酰胺(CTX)引起的免疫抑制作用,具有升高小鼠外周血中白细胞数的功能,而且能诱导外周血中TNF-α的升高。本实验从白色念珠菌细胞壁提取不溶性β-葡聚糖(Candida albicans insolubleβ-glucan,CAIBG),观察CAIBG对人外周血单核细胞(PBMC)的细胞活性以及TNF-α表达的影响,探讨其促进细胞因子分泌的情况及与抗肿瘤的相关性。【方法】首先从白色念珠菌细胞壁提取不溶性β-葡聚糖。常规培养PBMC,在培养液中加入不同浓度的CAIBG用改良MTT法检测12h、24h、36h时CAIBG对PBMC活性的影响,同时在加入不同浓度CAIBG后,分别作用不同时间用ELISA法检测TNF-α的含量,以确认PBMC中TNF-α与药物的量效关系和时效关系。培养宣威肺腺癌细胞,PBMC为效应细胞,人肺腺癌细胞系为靶细胞,设5∶1、10∶1和20∶1三个效靶比,随机分成四组:空白对照组、阴性对照组、阳性对照组、CAIBG组,观察各组PBMC对肺腺癌细胞的细胞毒作用,以及TNF-α的分泌情况。【结果】CAIBG浓度62.5ug/ml和125ug/ml作用与细胞12h、24h和36h时,PBMC活性明显升高,与空白对照组比较差异有统计学意义(P＜0.05)。CAIBG对PBMC能有效的促进TNF-α的分泌,并且有明显的时间依赖性,给药后随着时间的延长TNF-α的含量有所升高,在给药后第18h时TNF-α含量升高最明显,差异有统计学意义(P＜0.05)。CAIBG对PBMC分泌TNF-α有明显的剂量依赖性,给与不同的药物浓度后18h时TNF-α的含量均有所升高,但给药浓度为62.5ug/ml组TNF-α的含量升高最明显,差异有统计学意义(P＜0.05)。当效应细胞:靶细胞的比例为5∶1、10∶1和20∶1时,给与药物CAIBG浓度为62.5ug/ml的组PBMC毒作用高于空白对照组,差异有统计学意义(P＜0.05);而CAIBG组与顺铂组比较差异没有统计学意义(P＞0.05)。三种不同的效靶比时,给与药物CAIBG浓度为62.5ug/ml的组分泌TNF-α的含量与空白对照组、阴性对照组比较,差异均有非常显著性(P＜0.01);与顺铂组比较,差异有统计学意义(P＜0.05)。【结论】CAIBG可能是通过诱导PBMC产生TNF-α,从而杀伤肿瘤细胞,这可能是CAIBG抗肿瘤的重要机制之一,从而提示CAIBG是一种很有前途的生物反应调节剂,值得进一步研究和开发。
[Abstract]:[objective] Candida albicans cell wall insoluble 尾 -glucan (CAIBG) is a kind of biological response regulator (BRM), which has a wide range of biological activities, such as leukocytosis, anti-tumor, anti-virus and so on. CAIBG was successfully isolated from the cell wall of Candida albicans in the early stage of this study. It was proved by animal experiments that CAIBG could antagonize the immunosuppressive effect of cyclophosphamide (CTX) and increase the number of white blood cells in peripheral blood of mice. Moreover, it can induce the increase of TNF- 伪 in peripheral blood. In this study, Candida albicans insoluble 尾 -glucan-CAIBG was extracted from the cell wall of Candida albicans insoluble, and the effects of CAIBG on the cell activity and the expression of TNF- 伪 in human peripheral blood monocytes (PBMCs) were observed. To investigate the relationship between cytokine secretion and antitumor activity. [methods] the insoluble 尾 -glucan was extracted from the cell wall of Candida albicans. The effect of CAIBG on the activity of PBMC was detected by modified MTT method at 24 h and 36 h after conventional culture. At the same time, the content of TNF- 伪 was detected by Elisa at different time after adding different concentration of CAIBG. To confirm the dose-effect and time-effect relationship between TNF- 伪 and drug in PBMC. The lung adenocarcinoma cell line of Xuanwei was cultured as effector cell line and human lung adenocarcinoma cell line as target cell. The cells were randomly divided into four groups: blank control group, negative control group, positive control group and CAIBG group. The cytotoxic effect of PBMCs on lung adenocarcinoma cells and the secretion of TNF- 伪 were observed. [results] the activities of 62.5ug/ml and 125ug/ml were significantly increased at 12h and 36h, respectively. Compared with the control group, the difference was statistically significant (P < 0.05). CAIBG could effectively promote the secretion of TNF- 伪 in PBMC, and it was time-dependent, and the content of TNF- 伪 increased with time after administration. The content of TNF- 伪 increased most significantly at 18h after administration (P < 0.05). CAIBG was dose-dependent on the secretion of TNF- 伪 by PBMC, and the content of TNF- 伪 increased at 18h after administration of different drug concentrations. The concentration of TNF- 伪 in 62.5ug/ml group was significantly higher than that in control group (P < 0. 05). When the ratio of effector cells to target cells was 5: 1 / 10: 1 and 20:1, the toxicity of PBMCs in the group with drug 62.5ug/ml was higher than that in the control group. There was significant difference between CAIBG group and cisplatin group (P < 0.05), but there was no significant difference between CAIBG group and cisplatin group (P > 0.05). The levels of TNF- 伪 secreted by the 62.5ug/ml group were significantly different from those of the control group and the negative control group (P < 0. 01), and were significantly different from those of the cisplatin group (P < 0. 01), and compared with the control group (P < 0. 01) and the negative control group (P < 0. 01). [conclusion] CAIBG may kill tumor cells by inducing PBMC to produce TNF- 伪, which may be one of the important mechanisms of CAIBG anti-tumor. It is suggested that CAIBG is a promising biological response regulator, which is worthy of further study and development.
【学位授予单位】：昆明医学院
【学位级别】：硕士
【学位授予年份】：2008
【分类号】：R379

【引证文献】