本文選題：白色念珠菌不溶性β-葡聚糖 + PBMC�。� 參考：《昆明醫(yī)學院》2008年碩士論文

【摘要】： 【目的】白色念珠菌細胞壁不溶性β-葡聚糖(CAIBG)是一類生物反應調(diào)節(jié)劑(BRM),具有升白細胞、抗腫瘤、抗病毒等廣泛的生物學活性,本研究前期實驗已經(jīng)成功地從白色念珠菌細胞壁中分離出CAIBG,并通過動物實驗證實了CAIBG能對抗環(huán)磷酰胺(CTX)引起的免疫抑制作用,具有升高小鼠外周血中白細胞數(shù)的功能,而且能誘導外周血中TNF-α的升高。本實驗從白色念珠菌細胞壁提取不溶性β-葡聚糖(Candida albicans insolubleβ-glucan,CAIBG),觀察CAIBG對人外周血單核細胞(PBMC)的細胞活性以及TNF-α表達的影響,探討其促進細胞因子分泌的情況及與抗腫瘤的相關(guān)性。 【方法】首先從白色念珠菌細胞壁提取不溶性β-葡聚糖。常規(guī)培養(yǎng)PBMC,在培養(yǎng)液中加入不同濃度的CAIBG用改良MTT法檢測12h、24h、36h時CAIBG對PBMC活性的影響,同時在加入不同濃度CAIBG后,分別作用不同時間用ELISA法檢測TNF-α的含量,以確認PBMC中TNF-α與藥物的量效關(guān)系和時效關(guān)系。培養(yǎng)宣威肺腺癌細胞,PBMC為效應細胞,人肺腺癌細胞系為靶細胞,設5∶1、10∶1和20∶1三個效靶比,隨機分成四組:空白對照組、陰性對照組、陽性對照組、CAIBG組,觀察各組PBMC對肺腺癌細胞的細胞毒作用,以及TNF-α的分泌情況。 【結(jié)果】CAIBG濃度62.5ug/ml和125ug/ml作用與細胞12h、24h和36h時,PBMC活性明顯升高,與空白對照組比較差異有統(tǒng)計學意義(P＜0.05)。CAIBG對PBMC能有效的促進TNF-α的分泌,并且有明顯的時間依賴性,給藥后隨著時間的延長TNF-α的含量有所升高,在給藥后第18h時TNF-α含量升高最明顯,差異有統(tǒng)計學意義(P＜0.05)。CAIBG對PBMC分泌TNF-α有明顯的劑量依賴性,給與不同的藥物濃度后18h時TNF-α的含量均有所升高,但給藥濃度為62.5ug/ml組TNF-α的含量升高最明顯,差異有統(tǒng)計學意義(P＜0.05)。當效應細胞:靶細胞的比例為5∶1、10∶1和20∶1時,給與藥物CAIBG濃度為62.5ug/ml的組PBMC毒作用高于空白對照組,差異有統(tǒng)計學意義(P＜0.05);而CAIBG組與順鉑組比較差異沒有統(tǒng)計學意義(P＞0.05)。三種不同的效靶比時,給與藥物CAIBG濃度為62.5ug/ml的組分泌TNF-α的含量與空白對照組、陰性對照組比較,差異均有非常顯著性(P＜0.01);與順鉑組比較,差異有統(tǒng)計學意義(P＜0.05)。 【結(jié)論】CAIBG可能是通過誘導PBMC產(chǎn)生TNF-α,從而殺傷腫瘤細胞,這可能是CAIBG抗腫瘤的重要機制之一,從而提示CAIBG是一種很有前途的生物反應調(diào)節(jié)劑,值得進一步研究和開發(fā)。
[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- 偽 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- 偽 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- 偽 was detected by Elisa at different time after adding different concentration of CAIBG. To confirm the dose-effect and time-effect relationship between TNF- 偽 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- 偽 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- 偽 in PBMC, and it was time-dependent, and the content of TNF- 偽 increased with time after administration. The content of TNF- 偽 increased most significantly at 18h after administration (P < 0.05). CAIBG was dose-dependent on the secretion of TNF- 偽 by PBMC, and the content of TNF- 偽 increased at 18h after administration of different drug concentrations. The concentration of TNF- 偽 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- 偽 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- 偽, 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.
【學位授予單位】：昆明醫(yī)學院
【學位級別】：碩士
【學位授予年份】：2008
【分類號】：R379

【引證文獻】

相關(guān)碩士學位論文 前1條

1 陶佳;白色念珠菌細胞壁不溶性β-葡聚糖（β-glucan）抗腫瘤細胞的實驗研究[D];昆明醫(yī)學院;2008年

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本文編號：2071615