Myricetin alleviated immunologic contact urticaria and mast cell degranulation via the PI3K/Akt/NF-κB pathway.

Immunologic contact urticaria (ICU) is characterized by the wheal and flare reaction from direct contact with a chemical or protein agent, which involves a type I hypersensitivity mediated by allergen-specific immunoglobulin E (sIgE). Myricetin (Myr), a bioactive flavonoid, exhibits antiinflammatory activities. Our results showed that treatment with Myr could alleviate ICU symptoms, including a decrease in the number of wheals and scratching, and inhibit ear swelling in the IgE/DNFB-induced mice. The serum level of IgE, histamine, interleukin (IL)-4, TNF-α, and MCP-1 were reduced in Myr-treated mice. Myr also attenuated mast cells (MCs) degranulation and H-PGDS, TSLP, IL-33, PI3K, Akt, and NF-κB mRNA levels in ICU model. The IgE-mediated anaphylaxis mouse models demonstrated anti-allergic effects of Myr. In vitro analysis showed that Myr reduced IgE-induced calcium (Ca2+ ) influx, suppressed degranulation, and chemokine release in LAD2 cells (human primary mast cells). Myr can significantly inhibited PLCγ1, Akt, NF-κB, and p38 phosphorylation. In conclusion, the study demonstrated that Myr alleviate ICU symptoms and inhibit mast cell activation via PI3K/Akt/NF-κB signal pathway.

α-Linolenic acid attenuates pseudo-allergic reactions by inhibiting Lyn kinase activity.

BACKGROUND: Pseudo-allergic reactions are potentially fatal hypersensitivity responses caused by mast cell activation. α-linolenic acid (ALA) is known for its anti-allergic properties. However, its potential anti-pseudo-allergic effects were not much investigated. PURPOSE: To investigate the inhibitory effects of ALA on IgE-independent allergy in vitro, and in vivo, as well as the mechanism underlying its effects. METHODS/STUDY DESIGNS: The anti-anaphylactoid activity of ALA was evaluated in passive cutaneous anaphylaxis reaction (PCA) and systemic anaphylaxis models. Calcium imaging was used to assess intracellular Ca2+ mobilization. The release of cytokines and chemokines was measured using enzyme immunoassay kits. Western blot analysis was conducted to investigate the molecules of Lyn-PLCγ-IP3R-Ca2+ and Lyn-p38/NF-κB signaling pathway. RESULTS: ALA (0, 1.0, 2.0, and 4.0 mg/kg) dose-dependently reduced serum histamine, chemokine release, vasodilation, eosinophil infiltration, and the percentage of degranulated mast cells in C57BL/6 mice. In addition, ALA (0, 50, 100, and 200 µM) reduced Compound 48/80 (C48/80) (30 µg/ml)-or Substance P (SP) (4 µg/ml)-induced calcium influx, mast cell degranulation and cytokines and chemokine release in Laboratory of Allergic Disease 2 (LAD2) cells via Lyn-PLCγ-IP3R-Ca2+ and Lyn-p38/NF-κB signaling pathway. Moreover, ALA (0, 50, 100, and 200 µM) inhibited C48/80 (30 µg/ml)- and SP (4 µg/ml)-induced calcium influx in Mas-related G-protein coupled receptor member X2 (MrgX2)-HEK293 cells and in vitro kinase assays confirmed that ALA inhibited the activity of Lyn kinase. In response to 200 µM of ALA, the activity of Lyn kinase by (7.296 ± 0.03751) × 10-5 units/µl and decreased compared with C48/80 (30 µg/ml) by (8.572 ± 0.1365) ×10-5 units/µl. CONCLUSION: Our results demonstrate that ALA might be a potential Lyn kinase inhibitor, which could be used to treat pseudo-allergic reaction-related diseases such as urticaria.

Inhibitory function of Shikonin on MRGPRX2-mediated pseudo-allergic reactions induced by the secretagogue.

BACKGROUND: Mast cells (MCs) are crucial effectors in allergic disorders by secreting inflammatory mediators. The Mas-related G-protein-coupled receptor X2 (Mrgprx2) was shown to have a key role in IgE-independent allergic reactions. Therefore, potential drug candidates that directly target Mrgprx2 could be used to treat pseudo-allergic diseases. Shikonin, an active ingredient derived from Lithospermum erythrorhizon Sieb. et Zucc has been used for its anti-inflammatory properties since ancient China. PURPOSE: To investigate the inhibitory effects of Shikonin on IgE-independent allergy both in vitro and in vivo, as well as the mechanism underlying its effects. METHODS/STUDY DESIGNS: The anti-anaphylactoid activity of Shikonin was evaluated in PCA and systemic anaphylaxis models, Calcium imaging was used to assess intracellular Ca2+ mobilization. The release of cytokines and chemokines was measured using enzyme immunoassay kits. Western blot analysis was conducted to investigate the molecules of PLCγ-PKC-IP3 signaling pathway. The analytical method of surface plasmon resonance was employed to study the interaction between Shikonin and potential target protein Mrgprx2. RESULTS: Shikonin can suppress compound 48/80 (C48/80)-induced PCA, active systemic anaphylaxis, and MCs degranulation in mice in a dose-dependent manner. In addition, Shikonin reduced C48/80-induced calcium flux and suppressed LAD2 cell degranulation via PLCγ-PKC-IP3 signaling pathway. Moreover, Shikonin was found to inhibit C48/80-induced Mrgprx2 expression in HEK cells, displaying specific interactions with the Mrgprx2 protein. CONCLUSION: Shikonin could be a potential antagonist of Mrgprx2, thereby inhibiting pseudo-allergic reactions through Ca2+ mobilization.

Neohesperidin suppresses IgE-mediated anaphylactic reactions and mast cell activation via Lyn-PLC-Ca2+ pathway.

Mast cells play an essential role in IgE-FcεR1-mediated allergic diseases. Citrus aurantium is a prolific source of flavonoids with various biological activities, including anti-inflammatory, antioxidant, and anti-tumor efficacies. Neohesperidin is a novel flavonoid isolated from the leaves of C. aurantium. In this study, the anti-allergic and anti-inflammatory potentials of neohesperidin were investigated along with its molecular mechanism. The anti-anaphylactic activity of neohesperidin was evaluated through hind paw extravasation study in mice. Calcium imaging was used to assess intracellular Ca2+ mobilization. The levels of cytokines and chemokines were measured using enzyme immunoassay kits. Western blotting was used to explore the related molecular signaling pathways. Neohesperidin suppressed IgE-induced mast cell activations, including degranulation and secretion of cytokines and eicosanoids through inhibiting phosphorylation of Lyn kinase. Neohesperidin inhibited the release of histamine and other proinflammatory cytokines through a mast cell-dependent passive cutaneous anaphylaxis animal model. Histological studies demonstrated that neohesperidin substantially inhibited IgE-induced cellular infiltration and attenuated mast cell activation in skin tissue. In conclusion, our study revealed that neohesperidin could inhibit allergic responses in vivo and in vitro, and the molecule may be regarded as a novel agent for preventing mast cell-immediate and delayed allergic diseases.

[Clinical therapeutic effects of scraping therapy on allergic rhinitis of different syndromes].

OBJECTIVE: To compare the differences in the therapeutic effects on allergic rhinitis between scraping therapy and cetirizine and observe the differences in the clinical therapeutic effects of scraping therapy on the disease of different differentiated patterns/syndromes. METHODS: The included participants of allergic rhinitis were randomized into a western medication group and three scraping therapy groups, named a lung qi deficiency and cold group, a spleen qi deficiency group and a kidney yang deficiency group, 20 cases in each one. In the western medication group, cetirizine was prescribed for oral administration, 10 mg, once a day, totally for 4 weeks. In the scraping therapy groups, the scraping therapy was applied to the running courses of the large intestine meridian of hand-yangming, the governor vessel and the bladder meridian of foot-taiyang, focusing on Yingxiang (LI 20), Yintang (GV29), Hegu (LI 4) and Fengmen (BL 12). In the lung qi deficiency and cold group, the scraping focused on the lung meridian of hand-taiyin, Feishu (BL 13), Taiyuan (LU 9), Fengchi (GB 20) and Lieque (LU 7). In the spleen qi deficiency group, the scraping focused on the spleen meridian of foot-taiyin, Pishu (BL 20) and Zusanli (ST 36). In the kidney yang deficiency, the scraping focused on the kidney meridian of foot-shaoyin, Mingmen (GV 4) and Shenshu (BL 23). The scraping therapy was given once a week, 4 treatments as one session and 1 session required (4 weeks). Separately before treatment, after treatment and in 3-month follow-up visit, the total score of the main symptoms of allergic rhinitis (sneezing, runny nose, nasal obstruction and nasal itch) was observed and the clinical therapeutic effects were evaluated. RESULTS: The total symptom scores in the patients of the 4 groups after treatment were all reduced as compared with those before treatment (all P<0.05). After treatment and in the follow-up visit, the total symptom scores of the 3 scraping therapy groups were lower than those in the western medication group (all P<0.05), and the score in the lung qi deficiency and cold group was lower than those in the spleen qi deficiency group and the kidney yang deficiency group (all P<0.05). After treatment and in the follow-up visit, the therapeutic effects in the 3 scraping therapy groups were better than those in the western medication group (all P<0.05). CONCLUSION: The scraping therapy on the basis of the meridians and acupoints selection achieves the definite therapeutic effects on allergic rhinitis of different differentiated patterns/syndromes, which is better than cetirizine. This therapy achieves the much significant short-term and long-term therapeutic effects on allergic rhinitis differentiated as lung qi deficiency and cold.

Effect of Gua sha therapy on perimenopausal syndrome: a randomized controlled trial.

OBJECTIVE: This study aims to evaluate the effectiveness and safety of Gua sha therapy on perimenopausal symptoms, quality of life, and serum female hormones in participants with perimenopausal syndrome. METHODS: A prospective, randomized, controlled clinical trial was conducted at the First Affiliated Hospital of Nanjing University of Chinese Medicine in China. Eighty women with perimenopausal syndrome were recruited and randomized into an intervention group or a control group. Participants in the intervention group received 15-minute Gua sha treatment sessions once a week plus conventional treatment for 8 weeks, whereas participants in the control group received conventional treatment alone. The primary outcome was the change in perimenopausal symptoms and quality of life as obtained through the modified Kupperman Index (KI) and the Menopause-Specific Quality of Life. The secondary outcome was the change of serum female hormones including estrogen, follicle-stimulating hormone, and luteinizing hormone. RESULTS: Seventy-five out of 80 participants (93.8%) completed the study-38 in the intervention group and 37 in the control group. The baseline levels of demographic and outcome measurements were comparable between the two groups. After eight sessions of intervention, the reduction in the total modified KI score was, however, 16.32 ±â€Š4.38 in the intervention group and 11.46 ±â€Š5.96 in the control group, with a difference of 4.86 ±â€Š6.15 (P < 0.01) between the two groups. Also the reductions of hot flash/sweating, paresthesia, insomnia, nervousness, melancholia, fatigue, and headache were greater in the intervention group than in the control group (P < 0.05). The reduction in the total Menopause-Specific Quality of Life score was 17.87 ±â€Š3.84 in the intervention group and 13.62 ±â€Š7.40 in the control group, with a difference of 4.46 ±â€Š7.52 (P < 0.01) between the two groups. And the scores for vasomotor, psychosocial, and physical domains in the intervention group were significantly lower than those in the control group (P < 0.05). There were no significant differences in serum estrogen, follicle-stimulating hormone, and luteinizing hormone between the two groups. CONCLUSIONS: The results of this study suggest that Gua sha therapy was effective and safe in relieving perimenopausal symptoms and improving the quality of life in participants with perimenopausal syndrome. The therapy may serve as a promising, effective, nondrug treatment for perimenopausal syndrome in clinical work. Additional research is needed to better understand its effectiveness and examine its mechanism for treating perimenopausal syndrome.

Sinapine reverses multi-drug resistance in MCF-7/dox cancer cells by downregulating FGFR4/FRS2α-ERK1/2 pathway-mediated NF-κB activation.

BACKGROUND: Sinapine, an alkaloid derived from seeds of the cruciferous species, shows favorable biological properties, such as antioxidant and radio-protective activities. The inhibitory effect of sinapine on acquired chemoresistance in tumor cells and the underlying molecular mechanisms remain unknown. AIM: We examined the effect of sinapine on reversal of chemoresistance in Michigan Cancer Foundation 7 (MCF-7)/dox breast cancer cells. RESULTS: Combination treatment with sinapine and doxorubicin synergistically increased the cytotoxicity of doxorubicin in MCF-7/dox cells, as shown using a cell apoptosis assay. An accumulation assay demonstrated that sinapine increased the intracellular concentration of doxorubicin in a dose-dependent manner. Immunoblotting and real time polymerase chain reaction (RT-PCR) analysis showed that sinapine downregulated multi-drug resistance 1 (MDR1) expression. A significant correlation was observed between the expression of MDR1, phospho-factor receptor substrate (FRS), phospho-extracellular signal regulated kinase (ERK)1/2, and nuclear factor kappa B (NF-κB). Chromatin immunoprecipitation (ChIP) assay indicated that sinapine inhibited binding of the transcription factor NF-κB to the MDR1 promoter. CONCLUSIONS: Our findings indicated that sinapine played an important role in the downregulation of MDR1 expression through suppression of fibroblast growth factor receptor (FGFR)4/FRS2α-ERK1/2 mediated NF-κB activation in MCF-7/dox cancer cells.

[Observation on the therapeutic effect of scraping therapy on perimenopausal syndrome].

OBJECTIVE: To observe the clinical effects of scraping therapy on perimenopausal syndrome. METHODS: Twenty women with perimenopausal syndrome were treated with scraping therapy and the dorsal course of the Governor Vessell and the Urinary Bladder Meridian of Foot-Taiyang were scraped, especially on the Back-shu points and Ashi points. The clinical symptoms were observed and compared with a modified Kupperman score before and after treatment. RESULTS: In all the 20 patients, 3 cases were cured, 6 cases were markedly effective, 9 cases were effective, 2 cases were ineffective, and the total effective rate was 90.0%. The Kupperman total score after treatment of (10.4 +/- 7.5) was significantly lower than the score before treatment of (25.0 +/- 5.3) (P < 0.001), in which, hot flushes and sweating, insomnia, fatigue, paresthesia, anxiety/irritability, hypaphrodisia, urinary system infection, tinnitus, dizziness, memory deterioration and headache were eased significantly (P < 0.001, P < 0.05). CONCLUSION: The scraping therapy has a good clinical effect on perimenopausal syndrome and can significantly improve the clinical symptoms.