【摘要】：目的:探究LL-37在感染結(jié)核分枝桿菌(Mycobacterium tuberculosis,Mtb)的巨噬細(xì)胞(macrophage,Mφ)中對(duì)炎性因子分泌的影響及其抗炎作用。方法:(1)使用豆蔻酰佛波醇乙酯(PMA)將THP-1細(xì)胞誘導(dǎo)分化為具有吞噬能力的巨噬細(xì)胞(Mφ),用Mtb感染Mφ,建立細(xì)胞模型,用不同濃度的LL-37處理感染Mtb的巨噬細(xì)胞,并設(shè)置對(duì)照組。(2)實(shí)驗(yàn)分組:(1)正常對(duì)照組:THP-1+normal saline(NS);(2)Mtb組:THP-1+Mtb;(3)Mtb+5μg/ml LL-37組:THP-1+Mtb+5μg/ml LL-37;(4)Mtb+10μg/ml LL-37組:THP-1+Mtb+10μg/ml LL-37;(5)Mtb+20μg/ml LL-37組:THP-1+Mtb+20μg/ml LL-37。(3)不同時(shí)間點(diǎn)收獲細(xì)胞,提取RNA,用(Real-time PCR,RT-PCR)法檢測(cè)促炎因子IL-12p40、TNF-α及抗炎因子IL-4、IL-10 mRNA的表達(dá);用ELISA法測(cè)定以上各組細(xì)胞培養(yǎng)上清中細(xì)胞因子IL-12p40、TNF-α、IL-4、IL-10的分泌量。結(jié)果:與正常對(duì)照組相比,Mtb感染組各時(shí)間段IL-12p40、TNF-α、IL-4及IL-10的mRNA表達(dá)上調(diào),并且其分泌量顯著增加(P0.05);LL-37刺激組與Mtb組相比,各時(shí)間段外源性LL-37降低促炎因子IL-12p40、TNF-αmRNA的表達(dá),IL-12p40、TNF-α分泌下降(P0.05);而IL-4、IL-10 mRNA的表達(dá)升高,抗炎因子IL-4、IL-10分泌增加(P0.05)。結(jié)論:外源性的LL-37可抑制Mtb感染的巨噬細(xì)胞炎性細(xì)胞因子分泌,其作用效應(yīng)與LL-37的濃度、感染Mtb的時(shí)間相關(guān)。本研究為結(jié)核病的治療提供了新的視野和見(jiàn)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- 偽 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- 偽 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- 偽 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- 偽 mRNA and the secretion of IL-12p40 TNF- 偽 (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.
【學(xué)位授予單位】：重慶醫(yī)科大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2017
【分類(lèi)號(hào)】：R52

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