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目的:以THP-1细胞为载体,观察荆芥挥发油抗炎作用及与NLRP3炎症小体激活相关的调控作用机制。方法:MTS法测定荆芥挥发油对THP-1细胞的毒性作用;利用佛波酯(PMA)将THP-1细胞诱导分化为巨噬细胞,然后采用LPS与激活物(ATP、Nigericin、CPPD或Ca Cl2)进行造模,观察荆芥挥发油对NLRP3炎症小体激活的影响,ELISA法检测细胞上清中IL-1β、IL-18水平;RT-PCR法检测细胞中NLRP3、Caspase-1、IL-1β、IL-1α、IL-6 mRNA的表达。结果:荆芥挥发油浓度≤0.4 mg/mL时对THP-1巨噬细胞无毒;0.2 mg/mL荆芥挥发油对4种激活物诱导的THP-1巨噬细胞上清中IL-1β的高分泌具有显著抑制作用(P<0.05或P<0.01),且对ATP或Nigericin诱导下THP-1巨噬细胞NLRP3、Caspase-1、IL-1β、IL-1α、IL-6 mRNA的高表达具有显著下调作用(P<0.05或P<0.01)。结论:荆芥挥发油体外能抑制ATP、Nigericin、CPPD或Ca Cl2诱导的THP-1巨噬细胞IL-1β高分泌,抑制ATP或Nigericin诱导下THP-1巨噬细胞裂解液中NLRP3、Caspase-1、IL-1β、IL-1α、IL-6 mRNA的高表达,其抗炎作用的发挥与干预NLRP3炎症小体的激活有关。
OBJECTIVE: To investigate the anti-inflammatory effect of Nepeta volatile oil and the regulatory mechanism related to the activation of NLRP3 inflammasome using THP-1 cells as carrier. METHODS: Toxicity of Nepeta volatile oil to THP-1 cells was determined by MTS method. THP-1 cells were induced to differentiate into macrophages by phorbol ester (PMA), and then induced by LPS and activator (ATP, Nigericin, CPPD or Ca Cl2) to observe the effect of volatile oil of Nepeta on the activation of NLRP3 inflammasome. The levels of IL-1β and IL-18 in the supernatant of the cells were detected by ELISA. The expressions of NLRP3, Caspase-1 and IL- 1β, IL-1α, IL-6 mRNA expression. Results: Nepeta volatile oil concentration ≤ 0.4 mg / mL of THP-1 macrophages is non-toxic; 0.2 mg / mL Nepeta volatile oil of the four activators of THP-1 macrophage supernatant IL-1β high (P <0.05 or P <0.01), and high expression of NLRP3, Caspase-1, IL-1β, IL-1α and IL-6 mRNA in THP-1 macrophages induced by ATP or Nigericin had Significantly down-regulated (P <0.05 or P <0.01). Conclusion: Nepeta volatile oil could inhibit the secretion of IL-1β induced by ATP, Nigericin, CPPD or Ca Cl2 in THP-1 macrophages in vitro and inhibit the expression of NLRP3, Caspase-1 in THP-1 macrophage lysate induced by ATP or Nigericin , IL-1β, IL-1α, IL-6 mRNA expression, and its anti-inflammatory effects play an important role in the activation of NLRP3 inflammasome.