■桥本甲状腺炎(HT)是一种甲状腺自身免疫性疾病,其特征是淋巴细胞浸润和甲状腺破坏。夏枯草(PV)是一种传统的中草药,具有治疗HT的临床疗效。我们先前报道了PV在甲状腺细胞中的免疫调节作用;然而,PV的生物活性成分尚不清楚。本研究旨在阐明PV治疗HT的关键成分及其作用机制。
■网络药理学用于预测HT的关键PV成分。测试预测的成分以确定它们是否可以在人甲状腺细胞中发挥PV的免疫调节作用。有限的蛋白水解质谱(Lip-MS)用于探索人甲状腺细胞中与PV成分相互作用的蛋白质。使用微尺度热电泳结合测定来评估PV组分与靶蛋白的亲和力。
■从公共数据库中收集了具有192个组分靶标和3415个HT相关基因的11个PV组分。有了网络药理学,建立了一个“成分-目标-疾病”网络,其中包括槲皮素,木犀草素,山奈酚,Morin,和植物甾醇,β-谷甾醇被预测为HTPV的关键成分。在受刺激的原代人甲状腺细胞或Nthy-ori-31细胞中,关键成分抑制炎性细胞因子肿瘤坏死因子α(TNF-α)的基因表达,白细胞介素-6(IL-6),和干扰素-β(IFN-β),细胞凋亡,核因子κB(NF-κB)和干扰素调节因子3(IRF-3)的激活。热休克蛋白90α,A类,成员1(HSP90AA1),被Lip-MS鉴定为与PV中的类黄酮相互作用。Morin与HSP90AA1的亲和力最高(KD=122.74μM),其次是山奈酚(KD=168.53μM),木犀草素(KD=293.94μM),和槲皮素(KD=356.86μM)。
■槲皮素,木犀草素,山奈酚,Morin,β-谷甾醇在受刺激的人甲状腺细胞中再现了PV的抗炎和抗凋亡作用,这可能有助于PV在HT中的治疗效果。
UNASSIGNED: Hashimoto\'s thyroiditis (HT) is a thyroid autoimmune disease characterized by lymphocytic infiltration and thyroid destruction. Prunella vulgaris (PV) is a traditional Chinese herbal medicine with documented clinical efficacy in treating HT. We previously reported an immunoregulatory effect of PV in thyrocytes; however, the bioactive components of PV remained unclear. This study aimed to elucidate key components of PV for treating HT and their acting mechanisms.
UNASSIGNED: Network pharmacology was used to predict key PV components for HT. The predicted components were tested to determine whether they could exert an immunoregulatory effect of PV in human thyrocytes. Limited proteolysis-mass spectrometry (Lip-MS) was used to explore interacting proteins with PV components in human thyrocytes. Microscale thermophoresis binding assay was used to evaluate the affinity of PV components with the target protein.
UNASSIGNED: Eleven PV components with 192 component targets and 3415 HT-related genes were gathered from public databases. With network pharmacology, a \'component-target-disease\' network was established wherein four flavonoids including quercetin, luteolin, kaempferol, morin, and a phytosterol, β-sitosterol were predicted as key components in PV for HT. In stimulated primary human thyrocytes or Nthy-ori-31 cells, key components inhibited gene expressions of inflammatory cytokines including tumor necrosis factor α (TNF-α), interleukin-6 (IL-6), and interferon-β (IFN-β), cellular apoptosis, and activation of nuclear factor κB (NF-κB) and interferon regulatory factor 3 (IRF-3). Heat shock protein 90 alpha, class A, member 1 (HSP90AA1), was identified to interact with flavonoids in PV by Lip-MS. Morin had the highest affinity with HSP90AA1 (KD = 122.74 μM), followed by kaempferol (KD = 168.53 μM), luteolin (KD = 293.94 μM), and quercetin (KD = 356.86 μM).
UNASSIGNED: Quercetin, luteolin, kaempferol, morin, and β-sitosterol reproduced an anti-inflammatory and anti-apoptosis effect of PV in stimulated human thyrocytes, which potentially contributed to the treatment efficacy of PV in HT.