LL-37抑制结核分枝杆菌感染巨噬细胞炎性因子的分泌研究
发布时间：2018-10-08 13:43
【摘要】：目的:探究LL-37在感染结核分枝杆菌(Mycobacterium tuberculosis,Mtb)的巨噬细胞(macrophage,Mφ)中对炎性因子分泌的影响及其抗炎作用。方法:(1)使用豆蔻酰佛波醇乙酯(PMA)将THP-1细胞诱导分化为具有吞噬能力的巨噬细胞(Mφ),用Mtb感染Mφ,建立细胞模型,用不同浓度的LL-37处理感染Mtb的巨噬细胞,并设置对照组。(2)实验分组:(1)正常对照组:THP-1+normal saline(NS);(2)Mtb组:THP-1+Mtb;(3)Mtb+5μg/ml LL-37组:THP-1+Mtb+5μg/ml LL-37;(4)Mtb+10μg/ml LL-37组:THP-1+Mtb+10μg/ml LL-37;(5)Mtb+20μg/ml LL-37组:THP-1+Mtb+20μg/ml LL-37。(3)不同时间点收获细胞,提取RNA,用(Real-time PCR,RT-PCR)法检测促炎因子IL-12p40、TNF-α及抗炎因子IL-4、IL-10 mRNA的表达;用ELISA法测定以上各组细胞培养上清中细胞因子IL-12p40、TNF-α、IL-4、IL-10的分泌量。结果:与正常对照组相比,Mtb感染组各时间段IL-12p40、TNF-α、IL-4及IL-10的mRNA表达上调,并且其分泌量显著增加(P0.05);LL-37刺激组与Mtb组相比,各时间段外源性LL-37降低促炎因子IL-12p40、TNF-αmRNA的表达,IL-12p40、TNF-α分泌下降(P0.05);而IL-4、IL-10 mRNA的表达升高,抗炎因子IL-4、IL-10分泌增加(P0.05)。结论:外源性的LL-37可抑制Mtb感染的巨噬细胞炎性细胞因子分泌,其作用效应与LL-37的浓度、感染Mtb的时间相关。本研究为结核病的治疗提供了新的视野和见解。
[Abstract]:Aim: to investigate the effect of LL-37 on the secretion of inflammatory factors in macrophages infected with Mycobacterium tuberculosis (Mycobacterium tuberculosis,Mtb) and its anti-inflammatory effect. Methods: (1) THP-1 cells were induced to differentiate into phagocytic macrophages (M 蠁) by (PMA), and M 蠁 was infected with Mtb. The cell model was established. Macrophages infected with Mtb were treated with different concentrations of LL-37. (2) the control group: (1) normal control group: Mtb group: THP-1 Mtb; (3) Mtb 5 渭 g/ml LL-37 group: THP-1 Mtb 5 渭 g/ml LL-37; (4) Mtb 10 渭 g/ml LL-37 group: THP-1 Mtb 10 渭 g/ml LL-37; (5) Mtb 20 渭 g/ml LL-37 group: THP-1 Mtb 20 渭 g/ml LL-37. (3). The expression of proinflammatory factor IL-12p40,TNF- 伪 and anti-inflammatory factor IL-4,IL-10 mRNA was detected by Real-time PCR,RT-PCR method, and the secretion of IL-12p40,TNF- 伪 IL-4 IL-10 in the supernatant of above groups was determined by ELISA method. Results: compared with the normal control group, the mRNA expression of IL-12p40,TNF- 伪 IL-4 and IL-10 was up-regulated in the Mtb infected group, and the secretion of IL-4 and IL-10 was significantly increased (P0.05) compared with the Mtb group. Exogenous LL-37 decreased the expression of pro-inflammatory factor IL-12p40,TNF- 伪 mRNA and the secretion of IL-12p40 TNF- 伪 (P0.05), while the expression of IL-4,IL-10 mRNA increased and the secretion of anti-inflammatory factor IL-4,IL-10 increased (P0.05). Conclusion: exogenous LL-37 can inhibit the secretion of inflammatory cytokines in macrophages infected with Mtb, and its effect is related to the concentration of LL-37 and the time of Mtb infection. This study provides a new vision and insight for the treatment of tuberculosis.
【学位授予单位】：重庆医科大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R52
本文编号：2256968
[Abstract]:Aim: to investigate the effect of LL-37 on the secretion of inflammatory factors in macrophages infected with Mycobacterium tuberculosis (Mycobacterium tuberculosis,Mtb) and its anti-inflammatory effect. Methods: (1) THP-1 cells were induced to differentiate into phagocytic macrophages (M 蠁) by (PMA), and M 蠁 was infected with Mtb. The cell model was established. Macrophages infected with Mtb were treated with different concentrations of LL-37. (2) the control group: (1) normal control group: Mtb group: THP-1 Mtb; (3) Mtb 5 渭 g/ml LL-37 group: THP-1 Mtb 5 渭 g/ml LL-37; (4) Mtb 10 渭 g/ml LL-37 group: THP-1 Mtb 10 渭 g/ml LL-37; (5) Mtb 20 渭 g/ml LL-37 group: THP-1 Mtb 20 渭 g/ml LL-37. (3). The expression of proinflammatory factor IL-12p40,TNF- 伪 and anti-inflammatory factor IL-4,IL-10 mRNA was detected by Real-time PCR,RT-PCR method, and the secretion of IL-12p40,TNF- 伪 IL-4 IL-10 in the supernatant of above groups was determined by ELISA method. Results: compared with the normal control group, the mRNA expression of IL-12p40,TNF- 伪 IL-4 and IL-10 was up-regulated in the Mtb infected group, and the secretion of IL-4 and IL-10 was significantly increased (P0.05) compared with the Mtb group. Exogenous LL-37 decreased the expression of pro-inflammatory factor IL-12p40,TNF- 伪 mRNA and the secretion of IL-12p40 TNF- 伪 (P0.05), while the expression of IL-4,IL-10 mRNA increased and the secretion of anti-inflammatory factor IL-4,IL-10 increased (P0.05). Conclusion: exogenous LL-37 can inhibit the secretion of inflammatory cytokines in macrophages infected with Mtb, and its effect is related to the concentration of LL-37 and the time of Mtb infection. This study provides a new vision and insight for the treatment of tuberculosis.
【学位授予单位】：重庆医科大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R52
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