甲基强的松龙和香烟烟雾提取物对人类肺泡Ⅱ型上皮细胞（A549细胞）NF-κB、I-κBα、IL-6因子的影响
发布时间：2018-08-05 12:19
【摘要】：目的：香烟烟雾提取物(CSE)诱导人类肺泡Ⅱ型上皮细胞(A549)核因子-κB(NF-κB)、抑制蛋白-κBα(I-κ Bα)、白细胞介素-6(IL-6)的变化后，观察甲基强的松龙(Mp)对其的干预作用。方法：体外培养A549细胞系，根据目的和条件的不同将其分为三组：1)对照组(NS组)以无血清DMEM处理；2)刺激组(CSE组)仅以10％浓度CSE处理；3)治疗组(CSE+Mp组)同上CSE刺激后加入0.015％Mp处理。于不同时间段收集各组细胞裂解物后应用免疫组化法、免疫印迹法(Westernblot)，酶联免疫法(ELISA)等观察NF-κ B、I-κ B α的表达量与时间的关系及IL-6的变化。结果：组间有明显差异：1)对照组无变化；2)刺激组CSE可导致NF-κ B被活化，I-κ B α从处理后15分钟开始后减少至30分钟后最终消失，60-90分钟后恢复到刺激前水平，IL-6分泌增加；3)治疗组检测到NF-κ B表达量剧降，而I-κ Bα在15-30分钟内始终有本底表达，60-90分钟后恢复到刺激前水平，，IL-6分泌减少，组间比较发现I-κ Bα蛋白的表达水平明显低于对照组，高于刺激组，差异有统计学意义。(组间：F=19.633P＜0.01时间：F=14.88P＜0.01)。结论：甲基强的松龙能使CSE刺激后A549细胞NF-κ B活化减少，I-κ Bα蛋白表达增加，抑制NF κ B活性，减少了炎性细胞因子IL-6的分泌从而在呼吸道炎性疾病中起抗炎作用。
[Abstract]:Objective: cigarette smoke extract (CSE) induced the nuclear factor kappa B (NF- kappa B) of human alveolar type II epithelial cells (A549), inhibition of protein kappa B alpha (I- kappa B a) and interleukin -6 (IL-6), and observed the intervention effect of methylprednisolone (Mp) on it. Methods: the extracellular culture of A549 cell lines was divided into three groups according to the different purpose and conditions. 1) 1) the control group (group NS) was treated with serum-free DMEM; 2) the stimulation group (group CSE) was treated with only 10% concentration of CSE; 3) the treatment group (group CSE+Mp) was added to the 0.015%Mp treatment after the same CSE stimulation. The immunoblotting (Westernblot) and enzyme linked immunoassay (ELISA) were used to observe the NF- kappa B, I- kappa was observed in the treatment group (CSE+Mp group) after the same CSE stimulation. The relationship between the expression of B alpha and the time and the change of IL-6. Results: there were significant differences between the groups: 1) there was no change in the control group; 2) the stimulation group CSE could lead to the activation of NF- kappa B, and the I- kappa B alpha decreased from 15 minutes after treatment to 30 minutes after the treatment, and finally disappeared after 60-90 minutes, and the secretion of IL-6 increased in the 60-90 minutes, and 3) the treatment group detected NF. The expression of kappa B was dramatically reduced, and the expression of I- kappa B alpha always had the background expression in 15-30 minutes. After 60-90 minutes, the level of IL-6 secretion was reduced. The expression level of I- kappa B alpha protein was significantly lower than that of the control group. The difference was statistically significant. (group: F=19.633P < 0.01: F=14.88P < 0.01). Conclusion: Methylprednisolone can reduce the activation of NF- kappa B in A549 cells after CSE stimulation, increase the expression of I- kappa B alpha protein, inhibit the activity of NF kappa B, and reduce the secretion of inflammatory cytokines IL-6, so as to play an anti-inflammatory role in respiratory inflammatory diseases.
【学位授予单位】：新疆医科大学
【学位级别】：硕士
【学位授予年份】：2006
【分类号】：R363
本文编号：2165777
[Abstract]:Objective: cigarette smoke extract (CSE) induced the nuclear factor kappa B (NF- kappa B) of human alveolar type II epithelial cells (A549), inhibition of protein kappa B alpha (I- kappa B a) and interleukin -6 (IL-6), and observed the intervention effect of methylprednisolone (Mp) on it. Methods: the extracellular culture of A549 cell lines was divided into three groups according to the different purpose and conditions. 1) 1) the control group (group NS) was treated with serum-free DMEM; 2) the stimulation group (group CSE) was treated with only 10% concentration of CSE; 3) the treatment group (group CSE+Mp) was added to the 0.015%Mp treatment after the same CSE stimulation. The immunoblotting (Westernblot) and enzyme linked immunoassay (ELISA) were used to observe the NF- kappa B, I- kappa was observed in the treatment group (CSE+Mp group) after the same CSE stimulation. The relationship between the expression of B alpha and the time and the change of IL-6. Results: there were significant differences between the groups: 1) there was no change in the control group; 2) the stimulation group CSE could lead to the activation of NF- kappa B, and the I- kappa B alpha decreased from 15 minutes after treatment to 30 minutes after the treatment, and finally disappeared after 60-90 minutes, and the secretion of IL-6 increased in the 60-90 minutes, and 3) the treatment group detected NF. The expression of kappa B was dramatically reduced, and the expression of I- kappa B alpha always had the background expression in 15-30 minutes. After 60-90 minutes, the level of IL-6 secretion was reduced. The expression level of I- kappa B alpha protein was significantly lower than that of the control group. The difference was statistically significant. (group: F=19.633P < 0.01: F=14.88P < 0.01). Conclusion: Methylprednisolone can reduce the activation of NF- kappa B in A549 cells after CSE stimulation, increase the expression of I- kappa B alpha protein, inhibit the activity of NF kappa B, and reduce the secretion of inflammatory cytokines IL-6, so as to play an anti-inflammatory role in respiratory inflammatory diseases.
【学位授予单位】：新疆医科大学
【学位级别】：硕士
【学位授予年份】：2006
【分类号】：R363
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