1.Effect of Liangxue Tuizi Formula (凉血退紫方) on RAF/MEK/ERK Pathway in Skin Tissue and Serum NETs Biomarkers in Henoch-Schönlein Purpura Model Rats with Blood Heat Syndrome
Yingying JIANG ; Manxiang YANG ; Zhenhua YUAN ; Leying XI ; Mingyang CAI ; Diya MA ; Yifan LI ; Yuhang NIU ; Runze LIU ; Jiawen CAO ; Xilin CHEN ; Xianqing REN
Journal of Traditional Chinese Medicine 2025;66(23):2475-2483
ObjectiveTo investigate the potential mechanism of Liangxue Tuizi Formula (凉血退紫方, LXTZF) in treating Henoch-Schönlein Purpura (HSP) by examining its regulatory effect on neutrophil extracellular trap (NETs) dysregulation via the rapidly accelerated fibrosarcoma kinase (RAF)/mitogen-activated protein kinase (MEK)/extracellular signal-regulated kinase (ERK) signaling pathway. MethodsSeventy Wistar rats were randomly allocated into a blank control group (n=14) and a modeling group (n=56). Rats in the modelling group underwent an eight-week modelling period to establish HSP rat models with blood-heat syndrome via modified ovalbumin (OVA) induction method combined with oral administration of heat-property Chinese herbal medicine. Fifty successfully modeled rats were subsequently randomly divided into five groups (n=10 per group), model group, compound glycyrrhizin group, LXTZF group, RAF inhibitor group, and LXTZF + RAF agonist group. Additionally, 10 rats were selected from the original blank control group for the final experiment. From the 11th week of modelling, rats in the blank control group and the model group received 1 ml/(100 g·d) ultrapure water via oral administration, in addition to 0.5 ml/(kg·d) 0.9% sodium chloride solution via intraperitoneal injection. The LXTZF group and the compound glycyrrhizin group received 7.5 g/(kg·d) LXTZF granule suspension via gavage, 13.5 mg/(kg·d) compound glycyrrhizin suspension via gavage, respectively. The RAF inhibitor group received 1 mg/(kg·d) GW5074 suspension via intraperitoneal injection and ultrapure water via oral administration; the LXTZF + RAF agonist group received 7.5 g/(kg·d) LXTZF granule suspension via gavage and 1 mg/(kg·d) paclitaxel suspension via intraperitoneal injection. All administrations were performed once daily for 4 weeks. After intervention, skin tissue histopathology was examined by hematoxylin and eosin (H&E) staining, immunoglobulin A (IgA) deposition was assessed via immunofluorescence, serum levels of neutrophil elastase (NE), tumor necrosis factor-α (TNF-α), and vascular cell adhesion molecule-1 (VCAM-1) were measured using enzyme-linked immunosorbent assay (ELISA), serum myeloperoxidase (MPO) level was determined by a colorimetric assay; the mRNA expression levels of RAF, MEK, and ERK in skin tissue were detected by real-time quantitative polymerase chain reaction (RT-qPCR); and the protein expression of RAF, MEK, ERK, as well as phosphorylated MEK (p-MEK) and phosphorylated ERK (p-ERK), were analyzed by Western Blot. ResultsSkin tissue in the blank control group rats remained normal, whereas the model group exhibited neutrophil infiltration and haemorrhage with red blood cell rupture. In all drug intervention groups, neutrophil infiltration and haemorrhagic exudation reduced markedly, with LXTZF group demonstrating the most pronounced improvement. Compared with the blank control group, rats in the model group exhibited enhanced IgA fluorescence intensity in skin tissue, elevated serum levels of NE, MPO, TNF-α and VCAM-1, increased mRNA expression of RAF, MEK, ERK1 and ERK2, as well as heightened RAF protein levels and p-MEK/MEK and p-ERK/ERK ratios (P<0.05). Compared with the model group, the drug intervention groups exhibited reduced IgA fluorescence intensity in skin tissue, along with decreased serum levels of NE, MPO, TNF-α, and VCAM-1 (P<0.05). In LXTZF group and RAF inhibition groups, reduced mRNA expression of RAF, MEK, ERK1, and ERK2 was observed in rat skin tissue, alongside decreased RAF protein levels and reduced p-MEK/MEK and p-ERK/ERK ratios (P<0.05). Compared with LXTZF + RAF agonist group, the compound glycyrrhizin group, LXTZF group, and RAF inhibitior group exhibited reduced IgA fluorescence intensity in skin tissue, decreased serum NE, MPO, TNF-α, and VCAM-1 levels, and decreased MEK mRNA expression and p-MEK/MEK ratio (P<0.05). ConclusionThe potential mechanism by which LXTZF treats Henoch-Schönlein purpura with blood heat syndrome may involve blocking the RAF/MEK/ERK signaling pathway in skin tissue, and suppressing excessive formation of NETs, thereby reducing IgA deposition in dermal microvessels and attenuating systemic inflammatory responses.
2.A Review of Classic Formula Tingli Dazao Xiefeitang: Key Information Exploration and Ancient and Modern Applications
Yamin KONG ; Lyuyuan LIANG ; Jialei CAO ; Xuan WANG ; Liyuan CHEN ; Bingqi WEI ; Yujie CHANG ; Yihan LI ; Leying XI ; Bingxiang MA ; Wenli SHI
Chinese Journal of Experimental Traditional Medical Formulae 2024;30(24):208-214
Tingli Dazao Xiefeitang is a classic formula for treating lung abscesses and thoracic fluid retention, recognized throughout history and included in the Catalogue of Ancient Classic Formulas (Second Batch). This article employs bibliometric methods to investigate and analyze the source, origin, and key information of Tingli Dazao Xiefeitang, providing a theoretical basis for the development and application of this renowned formula. The results show that Tingli Dazao Xiefeitang first appears in Synopsis of the Golden Chamber (Jin Kui Yao Lue), where three clinical applications are recorded. The original text clearly states that this formula is used to treat lung abscesses and thoracic fluid retention, with symptoms such as inability to lie down due to wheezing, chest and rib fullness, facial edema, inability to rest due to thoracic fluid retention, and cough inversion. This aligns with findings from ancient application research. In later generations, the methods proposed by ZHANG Zhongjing were predominantly used for medication and formulation. Modern applications of Tingli Dazao Xiefeitang focus primarily on the respiratory system, with pleural effusion being the most common condition. For the preparation and decoction, a single dose of 5 g of Descurainiae Semen Lepidii Semen (dried and mature seeds of Lepidium apetalum) processed as stir-fried seeds and 36 g of Jujubae Fructus (dried and mature fruit of Ziziphus jujuba) prepared as raw materials were recommended. Then 600 mL of water was added and Jujubae Fructus was first boiled until the volume reduced to 400 mL. After filtering out the residue, the supernatant was retained and mixed with stir-fried Descurainiae Semen Lepidii Semen, followed by boiling to 200 mL, which should be consumed all at once. This research provides a theoretical basis for the development and application of this formula.
3.The Application of Sugen Theory in the Pathogenesis of Asthma
Qiongqiong XING ; Rongyi ZHOU ; Leying XI ; Yiwen YU ; Shuzi ZHANG ; Suping YU ; Rui LIN ; Xianqing REN
Journal of Nanjing University of Traditional Chinese Medicine 2024;40(6):645-652
Asthma is a common chronic respiratory disease characterized by repeated attacks and prolonged illness.Traditional Chinese medicine believes that the formation of Sugen is the core pathogenesis of repeated asthma attacks.By tracing the origin of Sug-en theory,summarizing the connotation of ancient asthma Sugen theory and the innovative understanding of modern medical scholars on asthma Sugen,this paper explores the potential connection between the traditional Chinese medicine Sugen theory and the pathogenesis of modern asthma,in order to provide new ideas and methods for the treatment and research of asthma.

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