苦参碱调控活动期强直性脊柱炎患者组蛋白甲基化酶EZH2表达干预Th17分化偏倚的机制

Mechanism of matrine regulating histone methylase EZH2 expression in patients with active ankylosing spondylitis to interfere with the imbalance of Th17 differentiation

目的:观察中药单体苦参碱对活动期强直性脊柱炎(AS)患者外周血单个核细胞(PBMC)组蛋白甲基化酶EZH2表达对Th17分化转录因子表达的抑制作用。方法:纳入2019年12月就诊于中国中医科学院广安门医院风湿病科门诊活动期AS患者10例,健康志愿者6名,梯度密度离心法分离PBMC,体外苦参碱单体干预24 h,Western Blot法检测苦参碱干预前后EZH2、RORc及H3K27me3蛋白表达情况,RT-PCR法检测苦参碱干预前后EZH2、RORc mRNA表达情况。结果:苦参碱干预24 h后,EZH2蛋白及mRNA表达较干预前显著升高(P<0.01),RORc蛋白及mRNA表达较干预前显著降低(P<0.01)。结论:中药单体苦参碱体外上调活动期AS-PBMC中EZH2表达,调控Th17分化及效应,是治疗AS炎症的可能有效药物之一。

Objective:To observe the inhibitory effect of matrine on the expression of histone methylase EZH2 in peripheral blood mononuclear cell(PBMC)of patients with active ankylosing spondylitis(AS)and on Th17 differentiation.Methods:This study included patients who visited the Department of Rheumatology of Guang’anmen Hospital,Chinese Academy of Traditional Chinese Medicine in December 2019.Among them,there were 10 active AS patients and 6 stable AS patients.PBMC was isolated by ficoll gradient density centrifugation.Subsequently,the extracted PBMC was subjected to 24-hour in vitro intervention with matrine.Western Blot method was used to detect the differences in the expression of proteins EZH2,RORc,and H3K27me3 before and after matrine intervention.Real-time quantitative PCR was used to detect the expression of EZH2 and RORc mRNA before and after matrine intervention.Results:After 24 hours of matrine intervention,the expression of EZH2 protein and mRNA was significantly increased compared to that before the intervention(P<0.01).The expression of RORc protein and mRNA was significantly lower than before the intervention(P<0.01).Conclusion:The traditional Chinese medicine monomer matrine upregulates the expression of EZH2 in active AS-PBMC in vitro.Meanwhile,it regulates Th17 differentiation and effects.Matrine is one of the possible effective drugs for treating AS inflammation.