目的 研究藁本内酯调控干扰素基因刺激蛋白(STING)/TANK结合激酶1(TBK1)/核因子-κB(NF-κB)信号对脂多糖(LPS)诱导的急性肺损伤小鼠模型肺组织损伤的影响.方法 将小鼠分成对照组、模型组(LPS肺损伤小鼠模型)、实验低剂量组(LPS肺损伤小鼠模型,20 mg·kg-1藁本内酯)、实验中剂量组(LPS肺损伤小鼠模型,60 mg·kg-1藁本内酯)、实验高剂量组(LPS肺损伤小鼠模型,100 mg·kg-1的藁本内酯)、DMXAA组(LPS肺损伤小鼠模型,100 mg·kg-1的藁本内酯、STING信号激动药DMXAA).用蛋白质印迹法检测STING、p-TBK1、TBK1、p-p65、p65蛋白的表达水平,用二喹啉甲酸(BCA)法检测肺泡灌洗液总蛋白,用瑞士吉姆萨染色计数肺泡灌洗液中炎性细胞数目,用酶联免疫吸附测定法检测肿瘤坏死因子-α(TNF-α)含量,用硫代巴比妥酸法检测肺组织中丙二醛(MDA)含量.结果 对照组、模型组和实验低、中、高剂量组及DMXAA组的STING蛋白相对表达水平分别为0.13±0.03、0.46±0.02、0.36±0.04、0.27±0.02、0.20±0.03 和 0.38±0.04,p-TBK1/TBK1 蛋白比值分别为 0.15±0.02、0.56±0.02、0.43±0.01、0.31±0.01、0.20±0.02 和0.30±0.02,p-p65/p65 蛋 白比值分别为 0.24±0.01、0.66±0.03、0.41±0.02、0.32±0.04、0.22±0.02和0.42±0.03,肺泡灌洗液中总蛋白含量分别为(173.79±15.44)、(304.21±19.29)、(243.59±17.60)、(212.51±18.24)、(187.11±11.59)和(241.69±30.12)μg·mL-1,巨噬细胞数目分别为(0.11±0.02)×108、(0.94±0.09)× 108、(0.72±0.08)× 108、(0.57±0.04)× 108、(0.41±0.04)×108和(0.71±0.08)×108cell·mL-1,模型组的上述指标与对照组相比,实验低、中、高剂量组的上述指标与模型组相比,DMXAA组的上述指标与实验高剂量组相比,在统计学上差异均有统计学意义(均P＜0.05).结论 藁本内酯抑制STING/TBK1/NF-κB信号减轻LPS诱导的急性肺损伤小鼠模型肺组织损伤.
Objective To study the effect of ligustilide regulating stimulator of interferon genes(STING)/Tank-binding kinase 1(TBK1)/nuclear factor-κB(NF-κB)signaling on lung tissue damage in mouse model of lipopolysaccharide(LPS)-induced acute lung injury.Methods Mice were divided into control group,model group(LPS lung injury mouse model),experimental-L group(LPS lung injury mouse model,20 mg·kg-1 ligustilide treatment),experimental-M group(LPS lung injury mouse model,60 mg·kg-1 ligustilide treatment),experimental-H group(LPS lung injury mouse model,100 mg·kg-1 ligustilide treatment),DMXAA group(LPS lung injury mouse model,100 mg·kg-1 ligustilide,STING signal agonist DMXAA treatment),Western blot was used to detect the STING,p-TBK1,TBK1,p-p65,p65 protein expression,bicinchoninic acid(BCA)method was used to detect total protein in bronchoalveolar lavage fluid,Swiss Giemsa staining was used to count the number of inflammatory cells in bronchoalveolar lavage fluid,the content of tumor necrosis factor-alpha(TNF-α)was detected by enzyme-linked immunosorbent assay(ELISA),and the content of malondialdehyde(MDA)in lung tissue was detected by thiobarbituric acid method.Results The relative expression levels of STING protein in the control,model,experimental-L,experimental-M,experimental-H groups,and DMXAA group were 0.13±0.03,0.46±0.02,0.36±0.04,0.27±0.02,0.20±0.03 and 0.38±0.04,p-TBK1/TBK1 protein ratios were 0.15±0.02,0.56±0.02,0.43±0.01,0.31±0.01,0.20±0.02 and 0.30±0.02,p-p65/p65 protein ratios were 0.24±0.01,0.66±0.03,0.41±0.02,0.32±0.04,0.22±0.02 and 0.42±0.03,the total protein contents in bronchoalveolar lavage fluid were(173.79±15.44),(304.21±19.29),(243.59±17.60),(212.51±18.24),(187.11±11.59)and(241.69±30.12)μg·mL-1;the number of macrophages were(0.11±0.02)×108,(0.94±0.09)×108,(0.72±0.08)×108,(0.57±0.04)×108,(0.41±0.04)×108 and(0.71±0.08)×108 cell·mL-1.The difference of the above indicators between model group and control group were all statistically significant(all P＜0.05);the difference between experimental-L,experimental-M,experimental-H groups and model group were all statistically significant(all P＜0.05);there were statistically significant difference between DMXAA group and experimental-H group(all P＜0.05).Conclusion Ligustolide inhibits STING/TBK1/NF-κB signaling to alleviate lung tissue damage in a mouse model of LPS-induced acute lung injury.
郑振羽;佘晖
福州市第二医院呼吸与危重症医学科,福建福州 350007
中医学
藁本内酯;急性肺损伤;干扰素基因刺激蛋白/TANK结合激酶1/核因子-κB信号;炎症;氧化损伤
ligustilide;acute lung injury;stimulator of interferon genes/Tank-binding kinase 1/nuclear factor-κB signaling;inflammation;oxidative damage
《中国临床药理学杂志》 2024 (011)
1588-1592 / 5
10.13699/j.cnki.1001-6821.2024.11.010
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