Shi Lu, An Yongcheng, Cheng Long, Li Yiyang, Li Huimin, Wang Chen, Lv Yinglan, Duan Yuhui, Dai Hongyu, He Changhao, Zhang Huilin, Huang Yan, Fu Wanxin, Wang ShengPeng, Zhao Baosheng, Wang Yitao, Zhao Yonghua
Department of Pharmacology, School of Chinese Materia Medica, Beijing University of Chinese Medicine, Beijing, 102488, China.
School of Life Sciences, Beijing University of Chinese Medicine, Beijing, 102488, China.
Chin Med. 2022 Jan 4;17(1):1. doi: 10.1186/s13020-021-00565-5.
Qingwei San (QWS), one of classic Chinese Medicine prescripts, has been widely used to treat stomach heat syndrome which manifests oral ulcer (OU), periodontitis and upper gastrointestinal bleeding for seven hundred years. However, the therapeutic effects of QWS on diabetic OU subjected to stomach heat syndrome are still ambiguous. In the study, we investigated the pharmacological mechanisms.
The main components of QWS aqueous extract were analyzed by LC-MS, and potential pathways of QWS targeting OU were predicted by network pharmacology. The db/db mice were administered with the decoction of dried Zingiber officinale Rosc. rhizome combined with NaOH cauterization to establish the model of diabetic OU subjected to stomach heat syndrome. Subsequently, the model mice were treated with QWS, and OU wound healing status were recorded. The pathological changes of gastric tissue and oral mucosa were evaluated using hematoxylin-eosin staining, and the morphology of collagen fibers in oral mucosa was assessed by Masson staining. The levels of thromboxane B (TXB), 6-Keto-prostaglandin F1α (6-keto-PGF1α), interleukin-1 β (IL-1β), IL-2, IL-6, tumor necrosis factor-α (TNF-α), β-endorphin (β-EP) and 5-Hydroxytryptamine (5-HT) were determined by ELISA assay. The protein expressions of Toll-like receptor 4 (TLR4), TNF receptor associated factor 6 (TRAF6), myeloid differentiation factor 88 (MyD88), inhibitor of NF-κB alpha (IκΒα), p-IκΒα and nuclear factor kappa-B (NF-κB) p65 were measured by Western Blotting.
A total of 183 compounds in QWS were identified by LC-MS, and identified 79 bioactive compounds corresponded to 269 targets and 59 pathways. QWS high-dose treatment significantly reduced the level of TXB and the ratio of TXB/6-keto-PGF1α. Meanwhile, it improved mucosal pathological morphology, and reduced the area of OU and local edema. Simultaneously, the levels of TNF-α, IL-1β, IL-6, IL-2 and 5-HT, and the expressions of TLR4, TRAF6, MyD88, p-IκΒα and NF-κB p65 were decreased.
QWS treatment facilitates the healing of OU, ameliorates pathological morphologies of gastric and oral mucosa and decreases the levels of pro-inflammatory cytokines in db/db mice subjected to stomach heat syndrome, whose mechanism may be associated with the inhibition of TLR4/MyD88/NF-κB signaling pathway to exert anti-inflammatory effects.
清胃散作为经典的中医方剂之一，七百年来一直被广泛用于治疗胃热证，其表现为口腔溃疡、牙周炎和上消化道出血。然而，清胃散对胃热证型糖尿病性口腔溃疡的治疗效果仍不明确。在本研究中，我们探究了其药理机制。
采用液相色谱-质谱联用（LC-MS）分析清胃散水提取物的主要成分，并通过网络药理学预测清胃散作用于口腔溃疡的潜在途径。采用干姜水煎剂联合氢氧化钠烧灼法建立胃热证型糖尿病性口腔溃疡模型。随后，给予模型小鼠清胃散治疗，并记录口腔溃疡愈合情况。采用苏木精-伊红染色评估胃组织和口腔黏膜的病理变化，采用Masson染色评估口腔黏膜胶原纤维形态。采用酶联免疫吸附测定（ELISA）法检测血栓素B（TXB）、6-酮-前列腺素F1α（6-keto-PGF1α）、白细胞介素-1β（IL-1β）、IL-2、IL-6、肿瘤坏死因子-α（TNF-α）、β-内啡肽（β-EP）和5-羟色胺（5-HT）的水平。采用蛋白质免疫印迹法检测Toll样受体4（TLR4）、肿瘤坏死因子受体相关因子6（TRAF6）、髓样分化因子88（MyD88）、核因子κB抑制因子α（IκΒα）、磷酸化IκΒα（p-IκΒα）和核因子κB（NF-κB）p65的蛋白表达。
通过LC-MS共鉴定出清胃散中的183种化合物，其中79种生物活性化合物对应269个靶点和59条通路。清胃散高剂量治疗显著降低TXB水平及TXB/6-keto-PGF1α比值。同时，改善了黏膜病理形态，减小了口腔溃疡面积和局部水肿。同时，TNF-α、IL-1β、IL-6、IL-2和5-HT水平以及TLR4、TRAF6、MyD88、p-IκΒα和NF-κB p65的表达均降低。
清胃散治疗可促进胃热证型db/db小鼠口腔溃疡的愈合，改善胃和口腔黏膜的病理形态，降低促炎细胞因子水平，其机制可能与抑制TLR4/MyD88/NF-κB信号通路发挥抗炎作用有关。
