Wang Xiao-Jun, Qi Yi-Ding, Guan Hao-Chen, Lin Hua-Gang, He Pei-Qing, Guan Kang-Wei, Fu Lei, Ye Mao-Qing, Xiao Jing, Wu Tao
Department of Traditional Chinese Medicine, Huadong Hospital Affiliated to Fudan University, Shanghai, China.
Shanghai Key Laboratory of Clinical Geriatric Medicine, Huadong Hospital Affiliated to Fudan University, Shanghai, China.
Front Pharmacol. 2021 May 4;12:665398. doi: 10.3389/fphar.2021.665398. eCollection 2021.
Gegen Qinlian decoction (GGQLD) is a typical traditional Chinese medicine (TCM) prescription documented in . Clinically, GGQLD has been utilized to manage the inflammatory symptoms of metabolic diseases and to protect against renal damage in China. In the present study, a hypothesis was proposed that the multi-target solution of GGQLD produced anti-inflammatory effects on ameliorating hyperuricemia (HUA). A total of 30 primary HUA patients receiving GGQLD treatment (two doses daily) for 4 weeks were selected. Then, differences in uric acid (UA) levels and expression of peripheral blood mononuclear cells (PBMCs) and urinary exosomes before and after treatment were analyzed. The therapeutic indexes for the active ingredients in GGQLD against HUA were confirmed through pharmacological subnetwork analysis. Besides, the HUA rat model was established through oral gavage of potassium oxonate and treated with oral GGQLD. In addition, proximal tubular epithelial cells (PTECs) were stimulated by UA and intervened with GGQLD for 48 h. Subsequently, RNA-seq, flow cytometry, and confocal immunofluorescence microscopy were further conducted to characterize the differences in UA-mediated inflammation and apoptosis of human renal tubular epithelial cells pre- and post-administration of GGQLD. In the meanwhile, quantitative real-time PCR (qPCR) was carried out to determine gene expression, whereas a western blotting (WB) assay was conducted to measure protein expression. Our network analysis revealed that GGQLD treated HUA via the anti-inflammatory and antiapoptotic pathways. Additionally, NLPR3 expression significantly decreased in PBMCs and urinary exosomes of HUA patients after GGQLD treatment. , GGQLD treatment alleviated HUA-induced renal inflammation, which was associated with decreased expression of NLRP3 inflammasomes and apoptosis-related mRNAs. Moreover, GGQLD promoted renal UA excretion by inhibiting the activation of GSDMD-dependent pyroptosis induced by NLRP3 inflammasomes and by reducing apoptosis via the mitochondrial apoptosis signaling pathway . This study indicates that GGQLD efficiently reduces inflammatory responses while promoting UA excretion in HUA. Our findings also provide compelling evidence supporting the idea that GGQLD protects against the UA-mediated renal tubular epithelial cell inflammation through the mitochondrial apoptosis signaling pathways. Taken together, these findings have demonstrated a novel therapeutic method for the treatment of HUA.
葛根芩连汤（GGQLD）是《 》中记载的一种典型的中药方剂。在中国，临床上GGQLD已被用于治疗代谢性疾病的炎症症状并预防肾脏损伤。在本研究中，提出了一个假设，即GGQLD的多靶点作用对改善高尿酸血症（HUA）具有抗炎作用。选取了30例接受GGQLD治疗（每日两剂）4周的原发性HUA患者。然后，分析治疗前后尿酸（UA）水平、外周血单个核细胞（PBMCs）和尿液外泌体表达的差异。通过药理子网分析确定了GGQLD中活性成分抗HUA的治疗指标。此外，通过口服氧嗪酸钾建立HUA大鼠模型并用口服GGQLD进行治疗。另外，用UA刺激近端肾小管上皮细胞（PTECs）并用GGQLD干预48小时。随后，进一步进行RNA测序、流式细胞术和共聚焦免疫荧光显微镜检查，以表征GGQLD给药前后UA介导的人肾小管上皮细胞炎症和凋亡的差异。同时，进行定量实时聚合酶链反应（qPCR）以确定基因表达，而进行蛋白质印迹（WB）分析以测量蛋白质表达。我们的网络分析表明，GGQLD通过抗炎和抗凋亡途径治疗HUA。此外，GGQLD治疗后HUA患者的PBMCs和尿液外泌体中NLRP3表达显著降低。 ，GGQLD治疗减轻了HUA诱导的肾脏炎症，这与NLRP3炎性小体和凋亡相关mRNA表达降低有关。此外，GGQLD通过抑制NLRP3炎性小体诱导的GSDMD依赖性焦亡的激活以及通过线粒体凋亡信号通路减少凋亡来促进肾脏UA排泄 。本研究表明，GGQLD在促进HUA患者UA排泄的同时有效减轻炎症反应。我们的研究结果还提供了令人信服的证据，支持GGQLD通过线粒体凋亡信号通路保护免受UA介导的肾小管上皮细胞炎症的观点。综上所述，这些发现证明了一种治疗HUA的新方法。
