BIOCHEMICAL PHARMACOLOGY
·
2020
·
IF 4.96
Nan Wang,Jue Wang,Yongjing Zhang,Yingnan Zeng,Shiling Hu,Haoyun Bai,Yajing Hou,Cheng Wang,Huaizhen He,Langchong He
Background Allergic asthma is a common inflammatory lung disease associated with complex pathogenesis. Mast cell (MC) is one of the key drivers of allergic asthma, Mas-related G protein-coupled receptor X2 (MRGPRX2) on the MC could mediate MC activation and trigger a pseudo-allergic reaction. Imperatorin (IMP), the main active compound of Radix Angelicae Dahuricae , has been reported to exert various pharmacological effects. In this study, we focused on the therapeutical mechanism of IMP on MRGPRX2-induced pseudo-allergy and allergic asthma. Methods We examined the effect of IMP on MRGPRX2 related mast cell activation in mouse peritoneal MC (MPMC), Human Laboratory of Allergic Disease 2 MCs (LAD2 cells) and Mrgprx2 -expressing HEK293 cells. Molecular docking and Surface plasmon resonance (SPR) were taken to reveal the binding character between IMP and MRPGRX2. MRGPRX2 downstream proteins were also detected by western blotting . IgE-independent responses was evaluated by using passive cutaneous anaphylaxis (PCA) and active systemic anaphylaxis (ASA) models. The therapeutic effect of IMP on asthma was evaluated by a lung inflammation mouse model which was induced by ovalbumin (OVA). Results IMP was found to reduce substance P (SP) induced calcium flux and suppressed degranulation of MC. SP can promote the phosphorylation of ERK and CamKII, which regulates the synthesis of inflammatory factors such as MIP-2 and TNF-α in MC. In vivo assays revealed that IMP can mitigate SP-induced mouse PCA and ASA. IMP could also mitigate lung inflammation in an OVA induced mice model by inhibiting MC activation in the lung tissue. Furthermore, IMP binds well to MRGPRX2 protein. The binding constant ( K D ) is 4.48 ± 0.49 × 10 −7 M. The data suggeste that IMP is a novel inhibitor of MRGPRX2 to treat allergic asthma.
研究领域:
药理学
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免疫学
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过敏
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呼吸系统疾病
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产品关联
DOI
PubMed
高影响因子
DYES AND PIGMENTS
·
2025
·
IF 4.2
Xiaoxue Mo,Yuexin Lv,Jiayu Lu,Yajing Hou,Hua Qian,Cheng Wang,Huaizhen He
MAS-related G protein-coupled receptor X2 (MRGPRX2) is a key target receptor and an important drug target for triggering pseudo-allergic reactions. Studying its ligands will help to understand the mechanism of pseudo-allergic reactions and discover anti-allergic drugs. Small-molecule fluorescent probes are widely used to study the localization of G protein-coupled receptors (GPCRs) on the cell membrane, the transport and internalization of GPCRs, and the interaction between receptors and ligands due to their high selectivity, sensitivity and safety. MRGPRX2 is one of GPCRs with an active hydrophobic ligand binding domain, so designing small-molecule fluorescent probes with environmental sensitivity is very important for the study of MRGPRX2 ligands and the visualization of this receptor. In this study, we selected the agonist of MRGPRX2 ( R )-ZINC-3573 as the recognition group and used different fluorophore skeleton structures as the luminescent group to synthesize a series of small-molecule fluorescent probes targeting MRGPRX2. Among them, PoX2-B with BODIPY FL as the fluorescent group had good photostability and environmentally sensitive properties to the hydrophobic ligand binding domain of MRGPRX2, and could be used for living cell imaging. Furthermore, PoX2-B could specifically recognize MRGPRX2, and it was expected to be used to screen effective ligands of MRGPRX2 in the future, so as to play a role in the development of anti-allergy drugs and the pathogenesis of allergic diseases.
研究领域:
药理学
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生物化学
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分子生物学
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DOI
高影响因子
American Journal of Translational Research
·
2025
·Vol. 17
·期 3
·2178
·
IF 1.6
Yu Zhang,Fang-Mei Liu,Cun-Yu Li,Xue-Jiao Leng,Yun-Feng Zheng,Guo-Ping Peng
Objective: Qingkailing Injection (QKLI) is a traditional Chinese medicine injection mainly used for sedation, heat clearing, and other treatments. However, recent clinical studies have shown a risk of pseudo-allergic reactions. The purpose of this study is to elucidate the underlying mechanism of QKLI-induced mast cell degranulation in Laboratory of Allergic Diseases 2 (LAD2) and to validate QKLI-induced activation of guinea pig IgE-independent allergic responses. Methods: Levels of β-hexosaminidase (β-Hex), histamine (His), and complement pathway-related indicators in guinea pigs and LAD2 cells were assayed using the Enzyme-linked Immunosorbent Assay (ELISA). The release rates of β-Hex and His from LAD2 cells were measured using the o-phthalaldehyde (OPA) fluorimetric method. The antagonist for complement component 3a (C3a) receptors, SB290157 and siRNAs were used to inhibit the C3a pathway and the Mas-related G-protein-coupled receptor X2 (MRGPRX2) pathway. The MRGPRX2 pathway and its downstream proteins were detected by Western Blot (WB). Results: The results show that QKLI significantly increased levels of β-Hex, His, C3a, complement component 5a (C5a), and terminal complement complex C5b-9 (SC5b-9) in guinea pigs, while levels of interleukin 4 (IL-4), interleukin 13 (IL-13), and interleukin 6 (IL-6) were unaffected. The C3a receptor inhibitor SB290157 significantly reduced levels of β-Hex and His. In LAD2 cells, QKLI increased the release rates of β-Hex and His in a time-dependent manner and decreased the phosphorylation of Extracellular Signal-regulated Kinase 1/2 (ERK1/2) proteins downstream of the MRGPRX2 pathway. The effective components of QKL, baicalin (BA) and geniposide (GE), individually enhance the allergic responses of LAD2 cells to some extent. However, the use of QKL is significantly superior to the individual use of its components. Conclusions: We found that QKLI induced pseudoanaphylaxis via an IgE-independent response in guinea pigs and through the MRGPRX2 pathway in human LAD2 cells. Among these, the main ingredients causing pseudoallergic reactions in QKLI were BA and GE. Our research contributes to a better understanding of the mechanisms underlying drug hypersensitivity reactions (DHRs).
研究领域:
药理学
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免疫学
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产品关联
DOI
PubMed
高影响因子
BIOMEDICINE & PHARMACOTHERAPY
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2022
·
IF 7.419
Cheng Wang,Yajing Hou,Shuai Ge,Jiayu Lu,Xiangjun Wang,Yuexin Lv,Nan Wang,Huaizhen He
Anaphylaxis is a severe systemic allergic reaction that exhibits multiple clinical symptoms. The Mas-related G protein-coupled receptor X2 (MRGPRX2) is recognized as a key cell receptor mediating allergic diseases and drug-induced anaphylactoid reactions. Thus, it has been a promising target for preventing and treating these reactions. Based on the potential activity of imperatorin and active structural feature of MRGPRX2, we first demonstrated that the synthetic imperatorin derivatives (IDs) could significantly inhibit MRGPRX2 agonist-induced degranulation and cytokine release in LAD2 cells, as well as alleviate local and systemic anaphylaxis in mice. The IC 50 value of the most promising compound is an order of magnitude lower than that of imperatorin. IDs were further identified to display anti-pseudo-allergic activity by binding MRGPRX2 with the tertiary nitrogen substructures, just liking the reported MRGPRX2-ligand. These results would propose evidence for discovery of agents for treating MCs-dependent allergic disorders.
研究领域:
免疫学
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药理学
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分子生物学
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产品关联
DOI
PubMed
高影响因子
ANALYTICAL BIOCHEMISTRY
·
2021
·
IF 3.365
Yuanyuan Ding,Xiaoqian Li,Qingpeng Gao,Xinyan Dong,Liyun Kong,Shengli Han,Tao Zhang,Langchong He
Mas-related G-protein-coupled receptor X2 (MRGPRX2) has recently been reported to be associated with anaphylaxis. Detection of MRGPRX2 levels in human peripheral blood might serve as a powerful tool for predicting the predisposition of patients to anaphylactic reactions. For rapid measurement of MRGPRX2, we established a paper-based double-antibody sandwich enzyme-linked immunosorbent assay (ELISA) using mouse monoclonal antibody and horseradish peroxidase (HRP)-labelled rabbit polyclonal antibody as capture antibody and detection antibody, respectively. We avoided chemical functionalization of the cellulose paper by introducing bovine serum albumin (BSA) to provide COOH and NH 2 groups for covalent immobilization of the capture antibody. Through amide condensation , a two-layer immobilization strategy was applied with BSA-BSA and BSA-capture antibody networks as the first and second layers, respectively. This strategy improved the quantity, activity and stability of the immobilized antibody . We then established a paper-based ELISA to detect MRGPRX2 in human peripheral blood. Our method is less laborious, easier to implement, and more cost-effective than conventional ELISA, while offering similar sensitivity, specificity, and accuracy. Therefore, it could serve as an innovative clinical point-of-care diagnostic tool, especially in areas that lack advanced clinical equipment.
研究领域:
医学
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免疫学
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生物化学
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DOI
高影响因子
JOURNAL OF AGRICULTURAL AND FOOD CHEMISTRY
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2017
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IF 3.154
Ting Wang,Xiao-Chun Hu,Zhi-Peng Cai,Josef Voglmeir,Li Liu
Recent progress in the relationship between carbohydrate cross-reactive determinants (CCDs) and allergic response highlights the importance of carbohydrate moieties in the innate immune system. Previous research pointed out that the protein allergen in Ginkgo biloba seeds is glycosylated, and the oligosaccharides conjugated to these proteins might also contribute to the allergy. The aim of this study was to analyze carbohydrate moieties, especially N-linked glycans, of glycoproteins from Ginkgo seeds originating from different places for detailed structures, to enable further research on the role played by N-glycans in Ginkgo-caused allergy. Results of monosaccharide composition and immunoblotting assays indicated the existence of N-glycans. Detailed structural elucidation of the N-glycans was further carried out by means of hydrophilic interaction ultraperformance liquid chromatography (HILIC-UPLC) and matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS). In total, 14 out of 16 structures detected by UPLC were confirmed by MALDI-TOF-MS and tandem mass spectrometry, among which complex-type N-glycans bearing Lewis A determinants and high-mannose-type N-glycans were identified from Ginkgo seeds for the first time. Precise quantification of N-glycans was performed by use of an external standard, and both the absolute amount of each N-glycan and the percentage of different types of N-glycan showed significant diversity among the samples without any pattern of geographic variation.
研究领域:
植物学
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免疫学
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生物化学
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产品关联
DOI
PubMed
高影响因子
Frontiers in Nutrition
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2025
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IF 5.1
Yanchu Li,Yuying Yang,Junyao Li,Rui Guo,Zhiqiang Niu,Weicheng Hu,Shucheng Liu,Shuai Wei
Introduction: Tropomyosin (TM) is the primary allergen in Litopenaeus vannamei, which usually causes allergic reactions that may be health or even life-threatening for consumers. Therefore, exploring the sensitization mechanism is of great significance for the prevention and treatment of tropomyosin allergy.Methods and results: In this study, TM sensitization models were using Balb/c mice, Caco-2 cells and RBL-2H3 cells to reveal the sensitization effect. The results of ELISA and RT-qPCR showed that TM can exacerbate the allergic reaction by reducing the mRNA expression of tight junction (TJ) proteins (such as ZO-1, claudin-3, Occludin) in the jejunum, destroying the intestinal barrier function, increasing the permeability, and promoting the release of inflammatory factors (such as IL-8, TNF-α) and histamine. The pathological results of intestinal tissue sections showed that TM also caused an increase in intestinal inflammatory infiltration in mice. RNA-seq analysis revealed that key genes (CCL2, HSP1A, GM-CSF, etc.) and PI3K/Akt signaling pathway were involved in the sensitization process. In vitro experiments were conducted to construct TM sensitized Caco-2 and RBL-2H3 cell models at a dose of 100 mg/mL. The results indicated that TM upregulated the expression of phosphorylated PI3K/ Akt and NFκB pathways in Caco-2 cells, further damaged the TJ structure of intestinal epithelial cells and promoted the release of inflammatory factors. The RBL-2H3 cell degranulation assay indicated that TM could directly stimulate the release of TNF-α from mast cells.Conclusion: The above experimental results indicated that PI3K/Akt signaling pathways play a crucial role in the induction of TM allergic responses, which provides a theoretical basis for the occurrence, development and prevention of TM allergy.
研究领域:
免疫学
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分子生物学
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药理学
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生物医学
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产品关联
DOI
PubMed
高影响因子
ANALYTICAL AND BIOANALYTICAL CHEMISTRY
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2012
·Vol. 404
·期 3
·887-893
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IF 3.778
Hu Xia,Wu Gong-ping,Zhang Meng-hui,Pan Shan-qing,Wang Rong-rong,Ouyang Jie-hu,Liu Jun-ge,Chen Zi-yuan,Tian Hong,Liu Dong-bo
Clinical definition and appropriate management of anaphylaxis is a clinical challenge because there is large variability in presenting clinical signs and symptoms. Monitoring of the metabolic status of anaphylaxis may be helpful in understanding its pathophysiological processes and diagnosis. The purpose of this study was to conduct GC–MS serum metabolic profiling of anaphylaxis animal models and search for potential biomarkers of anaphylaxis. Thirty-six guinea pigs were randomly divided into an ovalbumin group ( n = 12), a cattle albumin group ( n = 12), and a control group ( n = 12). The IgE level in the serum of the guinea pigs was evaluated by use of ELISA kits and the major metabolic changes in serum were detected by gas chromatography–mass spectrometry. Typical clinical symptoms appeared after the animals had been challenged with ovalbumin or cattle albumin. The IgE levels in serum of both model groups were significantly higher than those of the control group. Clustering trend of the three groups based on variables was observed and nine out of 858 metabolomic features were found to be significantly different between control group and model groups. Among the nine features, six features were tentatively identified as metabolites related to energy metabolism and signal transduction in anaphylaxis. In conclusion, GC–MS-based metabolic profiling analysis might be an effective auxiliary tool for investigation of anaphylaxis.
Chromatographic profile of the derivatized serum
研究领域:
临床医学
·
代谢组学
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DOI
高影响因子
JOURNAL OF DRUG DELIVERY SCIENCE AND TECHNOLOGY
·
2023
·
IF 5
Yupeng Feng,Peifu Xiao,Zixu Liu,Mei Li,Qingqing Wang,Tian Yin,Yu Zhang,Haibing He,Jingxin Gou,Yanjiao Wang,Xing Tang
Andrographolide (AND) and 14-deoxy-11, 12-dide-Hydroandrographolide (dehydroandrographolide, 14-DDA) are widely utilized traditional Chinese medicines for the treatment of inflammation. However, allergic reactions continue to be a concern. This research aimed to reduce allergic reactions and enhance the anti-inflammatory properties of 14-DDA by means of a liposome delivery system. To overcome the low encapsulation efficiency of conventional 14-DDA liposomes, a new 14-DDA derivative, dehydroandrographolide-divaline ester hydrochloride (DADH), was created by introducing valine groups at the C-3 and C-19 positions. DADH liposomes (DADH-lip) achieved high encapsulation efficiency (over 90%) via an ethanol injection method - pH gradient method. The lowest IC 50 value (16.34 μg/mL) of DADH-lip against P815 cells indicated that DADH-lip significantly reduced allergic reactions. In addition, DADH solution (DADH-sol) and DADH liposome decreased the amount of histamine released at both the high (1 mg/mL) and low (20 μg/mL) concentration dosages compared to the commercially available preparation. DADH-lip and DADH-sol also had significant effects in the LPS-induced RAW 264.7 macrophage model. In vivo anti-inflammatory results demonstrated that, compared to commercially available preparations, DADH-lip has a superior therapeutic effect and significantly reduces rat paw swelling. Collectively, DADH-lip with high encapsulation efficiency reduced allergic reactions significantly and enhanced therapeutic effect, suggesting that it may be a promising platform for inflammatory therapy.
研究领域:
药理学
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中药学
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纳米医学
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产品关联
DOI
高影响因子
APPLIED MICROBIOLOGY AND BIOTECHNOLOGY
·
2011
·Vol. 93
·期 1
·191-201
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IF 3.28
Wang Bingnan,Yang Xiufen,Zeng Hongmei,Liu Hua,Zhou Tingting,Tan Beibei,Yuan Jingjing,Guo Lihua,Qiu Dewen
PevD1, a novel protein elicitor from the pathogenic cotton verticillium wilt fungus, Verticillium dahliae , induced a hypersensitive response in tobacco plants. In this paper, the elicitor was purified and analyzed using de novo sequencing. The protein-encoding pevD1 gene consists of a 468-bp open reading frame that produces a polypeptide of 155 amino acids, with a theoretical molecular weight of 16.23 kDa. The sequence of elicitor protein PevD1 was matched to the genomic sequence (GenBank accession no. ABJE 01000445.1) of a putative protein from V. dahliae strain vdls.17, but a function had not yet been reported. The pevD1 gene was expressed in Escherichia coli , and the recombinant protein was characterized for its ability to confer systemic acquired resistance to tobacco mosaic virus (TMV). Recombinant PevD1-treated plants exhibited enhanced systemic resistance compared to control, including a significant reduction in the number and size of TMV lesions on tobacco leaves. The elicitor protein-induced hydrogen peroxide production, extracellular-medium alkalization, callose deposition, phenolics metabolism, and lignin synthesis in tobacco. Our results demonstrate that elicitor-PevD1 triggers defense responses in intact tobacco plants.
研究领域:
植物病理学
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分子生物学
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蛋白质化学
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DOI
PubMed
高影响因子
PHYTOTHERAPY RESEARCH
·
2021
·Vol. 35
·期 6
·3181-3193
·
IF 5.882
Rui Liu,Shiling Hu,Yuanyuan Ding,Jue Wang,Yuejin Wang,Jiapan Gao,Langchong He
Anaphylactoid reactions are potentially fatal allergic diseases caused by mast cells (MCs), which release histamine and lipid mediators under certain stimuli. Therefore, there is an urgent need to develop new drug candidates to treat anaphylactoid reactions. The MrgX2 receptor mediates anaphylactoid reactions that cause inflammatory diseases. Cortex dictamni is a Chinese herb used for treating allergy-related diseases; however, its active compound is still unknown and its mechanism of action has not yet been reported. The aim of this study was to screen the anti-anaphylactoid compound from C . dictamni extracts. An MrgX2/CMC-HPLC method was established for screening MrgX2-specific compounds retained from the alcohol extract of C . dictamni . A mouse model of hindpaw extravasation was used to evaluate the anti-anaphylactoid effect of this ingredient. Intracellular Ca 2+ mobilization was assessed using a calcium imaging assay. Enzyme immunoassays were performed to measure cytokine and chemokine release levels. The molecular signaling pathways were explored by western blotting. As a result, dictamnine was identified as an effective compound using the MrgX2/CMC method, which remarkably suppressed MC intracellular Ca 2+ mobilization and the release of de novo degranulated substances, and inhibited PKC-PLCγ-IP3R-associated protein signaling molecules. Hence, dictamnine is a novel therapeutic candidate for anaphylactoid reactions via MrgX2.
研究领域:
药理学
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免疫学
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中药学
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分子生物学
✓ 已验证
产品关联
DOI
PubMed
高影响因子
Toxicology Research
·
2021
·Vol. 10
·期 3
·467-475
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IF 3.524
Gao Jiapan,Che Delu,Du Xueshan,Zheng Yi,Jing Huiling,Wang Nan
Imidazolidinyl urea (IU) is used as an antimicrobial preservative in cosmetic and pharmaceutical products. IU induces allergic contact dermatitis, however, the mechanism has not yet been elucidated. Mas-related G protein-coupled receptor-X2 (MRGPRX2) triggers drug-induced pseudo-allergic reactions. The aims of this study were to determine whether IU activated mast cells through MRGPRX2 to further trigger contact dermatitis. Wild-type (WT) and KitW-sh/HNihrJaeBsmJNju (MUT) mice were treated with IU to observe its effects on local inflammation and mast cells degranulation in vivo. Laboratory of allergic disease 2 cells were used to detect calcium mobilization and release of inflammatory mediators in vitro. WT mice showed a severe local inflammatory response and contact dermatitis, whereas only slight inflammatory infiltration was observed in MUT mice. Thus, MRGPRX2 mediated the IU-induced activation of mast cells. However, histamine, a typical allergen, was not involved in this process. Tryptase expressed by mast cells was the major non-histaminergic inflammatory mediator of contact dermatitis. IU induced anaphylactic reaction via MRGPRX2 and further triggering non-histaminergic contact dermatitis, which explained why antihistamines are clinically ineffective against some chronic dermatitis.
研究领域:
免疫学
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皮肤科
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药理学
✓ 已验证
DOI
PubMed
高影响因子
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