基于网络药理学结合药动学及靶点验证探究黄芩清热除痹胶囊治疗类风湿关节炎质量标志物

To explore the quality markers of Huangqin Qingre Chubi capsule in the treatment of rheumatoid arthritis based on network pharmacology combined with pharmacokinetics and target verification

黄芩清热除痹胶囊(Huangqin Qingre Chubi capsule, HQC)作为临床治疗类风湿关节炎(rheumatoid arthritis, RA)的有效方剂,质量控制尚属空白。本研究拟通过网络药理学、药动学结合实验验证探究HQC治疗RA的质量标志物(quality markers, Q-marker)。通过构建“药效成分-靶点-通路”可视化网络,初步预测HQC治疗RA潜在质量标志物。建立风湿热痹证胶原诱导性关节炎(collagen-induced arthritis, CIA)大鼠模型,阐明HQC药效成分在大鼠疾病状态下的动态量化规律。建立肿瘤坏死因子-α (tumor necrosis factor-α, TNF-α)诱导的RA成纤维样滑膜细胞(MH7A)体外炎症模型,检测活性成分对鞘氨醇激酶1 (sphingosin kinase-1, Sphk1)、p-SphK1蛋白表达影响。网络药理学结果显示,黄芩苷、栀子苷、木犀草素、薏苡素和苦杏仁苷为HQC治疗RA的重要活性成分。定量分析结果进一步验证这5种成分的可测性。蛋白免疫印迹证实栀子苷和黄芩苷显著抑制Sphk1和p-SphK1蛋白表达。以上研究确定上述5种成分可作为HQC治疗RA的质量标志物。本实验经安徽中医药大学实验动物伦理委员会审核通过(批准号:AHUCM-rats-2021049),所有程序均严格按照动物使用和护理的原则进行。

As an effective prescription for the treatment of rheumatoid arthritis(RA),Huangqin Qingre Chubi capsule(HQC)is still blank in quality control.This study aims to explore quality markers(Q-markers)for HQC in the treatment of RA by integrating network pharmacology and pharmacokinetics.By constructing the visualization network of"pharmacodynamic ingredient-target-pathway",the potential Q-Marker of HQC treatment for RA was preliminatively predicted.A rat model of rheumatic heat obstruction syndrome collagene-induced arthritis(CIA)was established to elucidate the dynamic quantification law of pharmacodynamic components of HQC in the disease state of rats.To establish the inflammatory model of RA synovial fibroblasts(MH7A)induced by tumor necrosis factor-α(TNF-α)in vitro.The effects of active ingredients on protein expression of sphingosin kinase-1(Sphk1)and p-SphK1 were detected.The network pharmacological results showed that baicalin,geniposide,luteolin,coixol and amygdalin were the important active components of HQC treatment for RA.Quantitative analysis results further verified the measurability of these five components.The expression of Sphk1 and p-SphK1 was significantly inhibited by geniposide and baicalin by Western blotting.The above studies determined that the above 5 components could be used as Q-markers in the treatment of RA by HQC.This experiment was approved by the Experimental Animal Ethics Committee of Anhui University of Chinese Medicine(approval number:AHUCM-rats-2021049).All procedures were conducted in strict accordance with the principles of animal use and care.