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1.
Acta Pharmaceutica Sinica ; (12): 201-207, 2021.
Article in Chinese | WPRIM | ID: wpr-872599

ABSTRACT

The study is to investigate the effect of glaucocalyxin A (GLA) on mast cell-mediated anaphylaxis. The animal welfare and experimental process of this experiment followed the regulations of the Animal Ethics Committee of Yanbian University. BALB/c mice were used in the animal experiment and randomly divided into five groups, control group, model group, and GLA low, medium, and high dose groups (10, 20, and 40 mg·kg-1). Mice were sensitized by intradermal injection of anti-dinitrophenyl-immunoglobulin E (DNP-IgE) into the ears and challenged with a mixture of DNP-human serum albumin (HSA) and 4% evans blue into the tail veins to prepare an animal skin passive cutaneous anaphylaxis (PCA) model, which was collected from both ears for measurement of dye staining and histology. Rat peritoneal mast cells (RPMCs) were used in the cell experiment and divided into control, IgE + antigen (Ag), and IgE + Ag + GLA groups to determine histamine release as well as calcium influx levels. High-affinity IgE receptor (FcεRI)-mediated signaling pathway proteins and HMGB1/TLR4/NF-κB (high mobility group box 1/toll like receptor 4/nuclear transcription factor kappa B) signaling proteins were detected by Western blot. The results of animal experiments suggest that GLA inhibits PCA, reduces evans blue dye exudation, and reduces ear inflammation and ear thickness in mice. The results of cellular experiments suggested that GLA could reduce histamine release and calcium influx, and inhibit tumor necrosis factor-α (TNF-α), interleukin (IL)-4, IL-13, and IL-1β production; Western blot results showed that GLA inhibited FcεRI-mediated phosphorylation levels of spleen tyrosine kinase (Syk), Lck/Yes novel tyrosine kinase (Lyn), tyrosine kinase Fyn (Fyn), growth-factor receptor-bound protein 2 (Gab2), and phospholipase C (PLC) γ1, while GLA inhibited HMGB1/TLR4 signaling pathway to limit NF-κB p65 nuclear metastasis. The results indicate that GLA inhibits mast cell degranulation and attenuates allergic inflammation through the HMGB1/TLR4/NF-κB signaling pathway.

2.
Article in Chinese | WPRIM | ID: wpr-846665

ABSTRACT

Objective: To study the intervention effect of Gardenia jasminoside var. radicans and its main effective component of geniposide on the degranulation model of RBL-2H3 cells based on metabolomics. Methods: The changed metabolite profile of RBL-2H3 cells was detected by UPLC-QTOF-MS; PCA (principal component analysis) and OPLS-DA (orthogonal partial least squares discriminant analysis) in SIMCA software were used to select the potential biomarkers. Meanwhile, the clustering and heat map analysis for those potential biomarker levels were carried out by MEV software. Result: A total of 54 and 46 relevant biomarkers of G. jasminoside var. radicans and geniposide were selected, of which 31 biomarkers enriched in five disturbed metabolite pathways, including glycine, aspartic acid and glutamate metabolism, glutathione metabolism, histamine metabolism, energy metabolism, and nicotinamide metabolism pathways. Conclusion: G. jasminoside var. radicans and geniposide exerts the inhibitory effect on the degranulation model of RBL-2H3 cells by regulating histamine metabolism, oxidative stress and energy metabolism, and geniposide was one of the main efficacious substance basis of G. jasminoside var. radicans.

3.
Acupuncture Research ; (6): 299-304, 2020.
Article in Chinese | WPRIM | ID: wpr-844171

ABSTRACT

OBJECTIVE: To observe the effect of electroacupuncture (EA) on degranulation of intraperitoneal mast cells (MCs) and expression of mitogen-activated protein kinase (MAPK) signaling related proteins, tumor necrosis factor-α(TNF-α) and interleukin-6 (IL-6) in urticaria rats, so as to reveal its mechanisms underlying improvement of urticaria. METHODS: Thirty-two SD rats were randomly divided into control,model,EA and medication groups (n=8 in each group). The urticaria model was established by using passive cutaneous anaphylaxis (PCA) reaction method. EA (2 Hz /15 Hz, 1 mA) was applied to bilateral "Zusanli"(ST36), "Quchi "(LI11) and "Xuehai"(SP10) for 20 min,once daily for 7 consecutive days before antigen attack. Rats of the medication group were treated by gavage of Loratadine(1 mg•kg-1•d-1)for 7 days. The diameter of cutaneous Evan's blue spots was measured to evaluate the severity of PCA. Intraperitoneal fluid smears were prepared to observe the degranulation state of MCs. The contents of TNF-α and IL-6 in the intraperitoneal fluid were detected by ELISA, and the expression of extracellular signal-regulated kinase (ERK), phosphorylated (p)-ERK, c-Jun N-terminal kinase (JNK), p-JNK, P38MAPK and p-P38MAPK of the acquired intraperitoneal MCs was detected by Western blot. RESULTS: The diameter of cutaneous Evan's blue spot was significantly increased in the model group than that in the control group (P<0.01), and considerably decreased in both EA and medication groups compared with the model group(P<0.01). After modeling,the percentage of degranulated MCs, contents of TNF-α and IL-6, and expression levels of ERK, p-ERK, JNK, p-JNK, P38MAPK and p-P38MAPK were remarkably increased in the mo-del group than those in the control group (P<0.01, P<0.05). After the treatment, the percentage of degranulated MCs, contents of TNF-α and IL-6, and expression levels of p-ERK, JNK, p-JNK and p-P38MAPK were obviously decreased in both EA and medication groups relevant to the model group (P<0.01, P<0.05), while no significant changes were found in the expression of ERK in both EA and medication groups, and P38MAPK in the EA group. Compared with the model and EA groups, expression levels of P38MAPK were down-regulated in the medication group (P<0.05). CONCLUSION: EA can reduce skin allergic reaction in rats with urticaria, which may be related to its effects in inhibiting the degranulation of intraperitoneal MCs, down-regulating the expression of MAPK signaling-related proteins and the level of pro-inflammatory factors TNF-α and IL-6 in intraperitoneal MCs.

4.
Article in Chinese | WPRIM | ID: wpr-826658

ABSTRACT

OBJECTIVE@#To observe the effect of electroacupuncture (EA) preconditioning on the expressions of tyrosine kinase Lyn and spleen tyrosine kinase (Syk) in mast cells of subcutaneous loose connective tissue in the rats with urticaria and explore the potential biological mechanism of EA in the intervention of urticaria.@*METHODS@#A total of 32 SD rats were randomized into a blank group, a model group, an EA group and a positive medication group, 8 rats in each one. Except of the blank group, the passive cutaneous anaphylaxis (PCA) was adopted to prepare the model of urticaria in the rats of the rest three groups. In the EA group, EA was applied to bilateral "Quchi" (LI 11), "Xuehai" (SP 10) and "Zusanli" (ST 36), with disperse-dense wave, 2 Hz/15 Hz in frequency and 1 mA in current intensity, once daily, for 20 min each time, consecutively for 7 days. In the positive medication group, loratadine (1 mg•kg•d) was for intragastric administration, once daily, consecutively for 7 days. The samples were collected for index detection 30 min after PCA antigen challenge in the rats of each group. Spectrophotometer was adopted to determine the effusion quantity of Evans blue in the allergized site of skin. HE staining was used to observe the morphological changes in the allergized site of skin. Toluidine blue staining was provided to observe mast cell degranulation in subcutaneous loose connective tissue in the allergized site of skin. Immunohistochemistry was applied to determine the protein expressions of Lyn and Syk during degranulation of mast cells.@*RESULTS@#In the rats of the odel group, the eipdermis of allergized site was thickening, cells were disorganized in hierarchy and inflammatory cells were infiltrated largely in the dermis. In the positive medication group and the EA group, the epidermis was getting thin, cell arrangement was clear and the inflammatory cell infiltration was obviously alleviated as compared with the model group. Compared with the blank group, the OD value of skin dye effusion quantity, the degranulation rate of mast cells and the positive expressions of Lyn and Syk were all increased in the model group (<0.01). Compared with the model group, the OD value of skin dye effusion quantity, the degranulation rate of mast cells and the positive expressions of Lyn and Syk were all reduced in the EA group and the positive medication group (<0.01). Compared with the positive medication group, the degranulation rate of mast cells was increased significantly in the EA group (<0.01).@*CONCLUSION@#Electroacupuncture at "Quchi" (LI 11), "Xuehai" (SP 10) and "Zusanli" (ST 36) reduces vascular permeability and gives play to the role of anti-allergy by the way of regulating and controlling the degranulation of mast cells in the rats with urticaria and the effect mechanism of electroacupuncture may be related to the inhibition of protein expressions of Lyn and Syk in mast cells.


Subject(s)
Acupuncture Points , Animals , Connective Tissue , Metabolism , Electroacupuncture , Mast Cells , Metabolism , Random Allocation , Rats , Rats, Sprague-Dawley , Syk Kinase , Metabolism , Urticaria , Therapeutics , src-Family Kinases , Metabolism
5.
Article in Chinese | WPRIM | ID: wpr-873311

ABSTRACT

Objective::To study the effect of Yupingfeng granule on the degranulation of skin mast cells in chronic urticaria (CU) rats and the intervention mechanism of interleukin-23(IL-23), interleukin-17(IL-17) inflammation axis. Method::Totally 60 SPF SD rats were selected and randomly divided into normal group (normal saline), model group (normal saline), and loratadine group (0.9 mg·kg-1·d-1), high-dose Yupingfeng granules group (4.05 g·kg-1·d-1), middle-dose group (2.7 g·kg-1·d-1), low-dose group (1.35 g·kg-1·d-1). The CU rat model was reproduced through intraperitoneal injection of ovalbumin with aluminum hydroxide suspension and DTP vaccine. Histopathological changes of rat skin were observed by hematoxylin-eosin (HE) staining. Degranulation of mast cells in rat skin was determined by toluidine blue staining. IL-23 and IL-17 protein expressions in skin tissue were determined by immunohistochemistry (IHC). IL-23 and IL-17 mRNA transcription levels in skin tissue were determined by real-time fluorescence quantitative polymerase chain reaction (Real-time PCR). Result::Yupingfeng granules can significantly alleviate the pathological manifestations of dermal edema, collagen beam distance, inflammatory cell infiltration of CU rats, and reduce the degranulation reaction of skin tissue mast cells in CU rats. The IL-23, IL-17 mRNA and protein expressions of the skin of model group were significantly increased compared with the normal group (P<0.05, P<0.01). Compared with the model group, Yupingfeng granules can significantly down-regulate IL-23 mRNA and protein expressions of CU rats (P<0.05, P<0.01). Yupingfeng granules had no significant regulatory effect on IL-17. Conclusion::Yupingfeng granule can significantly reduce the degranulation of mast cells in skin tissue of CU rats, and improve the pathological manifestations, such as dermal edema, serous exudation and inflammatory cell infiltration. The mechanism may be related to inhibiting the secretion of IL-23 pro-inflammatory cytokines and improving CU lesions.

6.
Article in Chinese | WPRIM | ID: wpr-872758

ABSTRACT

Objective:To observe the influence of Chang'an Ⅰ prescription drug-containing serum on IgE-mediated RBL-2H3 cell degranulation model, and explore the mechanism of Chang'an Ⅰ prescription in inhibiting RBL-2H3 activation degranulation and releasing inflammatory mediators with v-yes-1 Yanaguchi sarcoma viral related oncogene homolog (Lyn)/spleen tyrosine protein kinase (Syk)/mitogen-activated protein kinase (MAPK) signal pathway. Method:Preparation for Chang'an Ⅰ prescription serum. Animal group, SD male rats were randomly divided into Chang'an Ⅰ prescription serum high, medium, low dose, and blank control groups with 10 rats in each group. Dosage: 10 mL·kg-1 distilled water was given to blank control group, while Chang'an Ⅰ prescription serum high, medium and low dose groups were respectively given to the Chang'an Ⅰ prescription concentrated crude drug with concentration of 1.15,2.30,4.60 g·kg-1, respectively once a day for 7 days continuously and then blood was taken from aorta ventralis and centrifuged. Ketotifen as the positive control drug. Mast cells are counted with toluidine blue staining. Cellular release of β-aminohexose was detected by colorimetric method. Contents of MCT, TNF-α, MCP-1 and histamine were measured by enzyme-linked immunosorbent assay (ELISA) kits, Lyn/Syk/MAPK protein levels were detected by immunoblotting. Result:For cell activation and degranulation, compared with the blank control group, the model group had more cell degranulation (P<0.05), compared with model group, the cell degranulation rate of each dose group of Chang'an Ⅰ prescription decreased (P<0.05). The release rate of β-hexosamine in each dose group of Chang'an Ⅰ prescription decreased significantly (P<0.01). For the release of active mediators, compared with the blank control group, the contents of histamine, MCT, TNF-α and MCP-1 all increased in the model group (P<0.01), compared with the model group, the contents in each dose group of Chang'an Ⅰ prescription all decreased significantly (P<0.01). Compared with the normal group, the phosphorylation levels of Lyn and Syk, extracellular regulatory protein kinase 1/2(ERK1/2), c-Jun N-terminal kinase (JNK), and mitogen-activated protein kinase p38 increased in the model group (P<0.05). Compared with the model group, the Lyn, Syk and ERK1/2, JNK and p38 protein phosphorylation levels reduced in Chang'an Ⅰ prescription group (P<0.05). Conclusion:Chang'an Ⅰ prescription drug-containing serum down-regulates the phosphorylation levels of proteins Lyn, Syk, and ERK1/2, JNK, and p38, inhibits RBL-2H3 cell activation and degranulation, reduces the release of cytokines and chemokines, such as histamine, MCT, TNF-α and MCP-1, it may be one of its mechanisms for treating IBS-D visceral hypersensitivity.

7.
Article in Chinese | WPRIM | ID: wpr-773650

ABSTRACT

This paper was aimed to establish screening methods of anaphylactoid reaction caused by safflower yellow for injection based on RBL-2 H3 cell degranulation model and mice model for acute anaphylactoid reaction,and evaluate the hypersensitivity caused by safflower yellow for injection from different batches. An in vitro cell model was used to keep the cells stimulated for an hour with different batches of safflower yellow for injection as the drug group,serum-free MEM medium as negative control group and 30 mg·L-1 C48/80 as positive control group respectively. The supernatant was then absorbed,and neutral red staining technique was used to detect the effect of safflower yellow injection on the degranulation of RBL-2 H3 cells with the positive cell rate of degranulation as the indicator.An in vivo model was established to validate the experimental results,and mice model for acute anaphylactoid reaction and ELISA method were adopted to detect the plasma histamine content,and screen the hypersensitivity caused by safflower yellow for injection at the animal level by using plasma histamine content as a test index. The results of the neutral red staining experiments showed that the positive control C48/80 could cause cell degranulation,and most of the cells were deeply stained. There was significant difference in positive cell rate between different batches of safflower yellow and positive control group. In the mice model for acute anaphylactoid reaction,it was found that the positive control C48/80 significantly increased the histamine content in the plasma of mice,while the safflower yellow in each batch did not cause a significant increase in plasma histamine( P<0. 000 1). The mechanism of anaphylactoid reaction is relatively complicated. This study was mainly based on the release of histamine and other active substances by degranulation of mast cells. No significant degranulation reaction of RBL-2 H3 cells induced by safflower yellow for injection was detected,nor was the plasma histamine level significantly increased in mice from the in vitro and in vivo aspects.


Subject(s)
Anaphylaxis , Animals , Cell Degranulation , Cells, Cultured , Chalcone , Histamine , Blood , Mast Cells , Mice
8.
Chinese Journal of Dermatology ; (12): 241-247, 2019.
Article in Chinese | WPRIM | ID: wpr-745772

ABSTRACT

Objective To evaluate the clinical efficacy of autologous whole blood injections (AWBI) combined with antihistamines for the treatment of patients with refractory chronic spontaneous urticaria and positive autologous serum skin test (ASST),to evaluate its effect on the expression of the high-affinity IgE receptor (FcεR Ⅰ) and CD63 on basophils,and to analyze the possible mechanism underlying the treatment of ASST-positive chronic urticaria with AWBI.Methods Eighty patients with ASST-positive chronic intractable urticaria were enrolled from Department of Dermatology,The First Hospital Affiliated to Army Medical University between November 2017 and June 2018,and randomly and equally divided into two groups by a random number table:AWBI group and control group were both conventionally treated with oral loratadine and ebastine,and AWBI group were additionally treated with AWBI once a week for 12 sessions.Before the treatment and after 12-week treatment,urticaria activity score of 7 days (UAS7) and dermatology life quality index (DLQI) in the two groups were evaluated.Among 30 patients in the AWBI group,flow cytometry was performed to determine the expression of FcεRⅠ and CD63 on the basophils in the peripheral blood at the baseline,weeks 4,8 and 12 after the initial treatment.Statistical analysis was carried out with GraphPad Prism 7.00 software by t test for the comparison of UAS7 or DLQI scores,Mann-Whitney U test for the comparison of FcεR Ⅰ α expression,paired Wilcoxon signed rank test for comparing FceR Ⅰ α or CD63 expression between two different time points,and Spearman correlation analysis for analyzing the correlation between FcεR Ⅰ α and CD63 expression.Results Before the treatment,no significant differences in UAS7 or DLQI scores were observed between the AWBI group and control group (UAS7:27.15 ± 4.53 vs.26.90 ± 5.22;DLQI:16.88 ± 6.01 vs.17.08 ± 6.79;both P > 0.05).After 12-week treatment,UAS7 and DLQI scores both significantly decreased in the two groups compared with those before the treatment (all P < 0.01),and were significantly lower in the AWBI group than in the control group (UAS7:14.25 ± 7.56 vs.19.93 ± 6.32;DLQI:8.48 ± 4.15 vs.13.93 ± 5.43;both P < 0.01).At the baseline,weeks 4,8 and 12 after the initial treatment,the fluorescence intensities of FcεR Ⅰα on basophils (M [P25,P75]) in the AWBI group were 22 532 (16 740,29 220),16 911 (10 240,21 816),13 282 (7 600,16 848) and 11 466 (7 161,14 578) respectively,and the proportions of CD63+ basophils induced by ASST-positive serum (M [P25,P75]) in the AWBI group were 35.25% (26.75%,49.13%),25.95% (19.37%,37.54%),13.57% (7.79%,19.57%) and 9.87% (6.43%,16.52%) respectively.At week 4 after the initial treatment,the expression of FcεR Ⅰα and CD63 on basophils in the AWBI group both significantly decreased compared with those at the baseline (both P < 0.01),but significantly increased compared with those at week 8 (both P < 0.01).The changes in FcεR Ⅰ α expression from baseline to week 4,from week 4 to week 8,and from week 8 to week 12 were positively correlated with the changes in CD63 expression induced by ASST-positive serum (r =0.364,0.422,0.455,respectively,all P < 0.05).Conclusion AWBI combined with antihistamines can improve the clinical symptoms of ASST-positive refractory chronic urticaria,likely by affecting the expression of FcεR Ⅰ and CD63 on basophils.

9.
Article in English | WPRIM | ID: wpr-716686

ABSTRACT

Asthma and autoimmune diseases both result from a dysregulated immune system, and have been conventionally considered to have mutually exclusive pathogenesis. Autoimmunity is believed to be an exaggerated Th1 response, while asthma with a Th2 underpinning is congruent with the well-accepted Th1/Th2 paradigm. The hypothesis of autoimmune involvement in asthma has received much recent interest, particularly in the adult late-onset non-atopic patients (the “intrinsic asthma”). Over the past decades, circulating autoantibodies against diverse self-targets (beta-2-adrenergic receptors, epithelial antigens, nuclear antigens, etc.) have been reported and subsequently dismissed to be epiphenomena resulting from a chronic inflammatory condition, primarily due to lack of evidence of causality/pathomechanism. Recent evidence of ‘granulomas’ in the lung biopsies of severe asthmatics, detection of pathogenic sputum autoantibodies against autologous eosinophil proteins (e.g., eosinophil peroxidase) and inadequate response to monoclonal antibody therapies (e.g., subcutaneous mepolizumab) in patients with evidence of airway autoantibodies suggest that the role of autoimmune mechanisms be revisited. In this review, we have gathered available reports of autoimmune responses in the lungs, reviewed the evidence in the context of immunogenic tissue-response and danger-associated molecular patterns, and constructed the possibility of an autoimmune-associated pathomechanism that may contribute to the severity of asthma.


Subject(s)
Adult , Antigens, Nuclear , Asthma , Autoantibodies , Autoimmune Diseases , Autoimmunity , Biopsy , Eosinophils , Humans , Immune System , Immunoglobulin G , Lung , Neutrophils , Sputum
10.
Chinese Journal of Pathophysiology ; (12): 1081-1088, 2018.
Article in Chinese | WPRIM | ID: wpr-701243

ABSTRACT

AIM:To detect the expression of CBir1 in the serum and colon tissue and mast cell degranulation in the tissue of 2,4,6-trinito-benzene-sulfonic acid ( TNBS)-induced colitis in mice with different interventions. ME-THODS:SPF male BALB/c mice were randomized into 6 groups (12 mice in each group):normal control group, normal saline group, 50% alcohol group, 50% alcohol+TNBS group, 50% alcohol+TNBS+lipopolysaccharide (LPS) +ovalbu-min (OVA) group and 50% alcohol+TNBS+ketotifen group. Corresponding treatment was given to each group, and the disease activity index (DAI) of the mice was evaluated. The mice were sacrificed on day 22 after treatment. The colon tis-sues were evaluated by histological index (HI) scoring. Serum concentrations of anti-CBir1, mast cell tryptase (MCT) and histamine were measured by ELISA. The expression of CBir1, toll-like receptor 5 (TLR5) and MCT in the colon tissues was detected by immunohistochemistry. RESULTS:Compared with the normal control group, the DAI score, HI score and CBir1, anti-CBir1, MCT, TLR5, histamine concentrations in colon tissues and serum were all significantly higher in 50% alcohol+TNBS group, 50% alcohol+TNBS+ketotifen group and 50% alcohol+TNBS+LPS+OVA group (P<0.05). The DAI score, HI score and anti-CBir1, CBir1, MCT, histamine levels in 50% alcohol+TNBS group were lower than those in 50% alcohol+TNBS+LPS+OVA group (P<0.05). The DAI score, HI score and anti-CBir1, TLR5, hista-mine, CBir1 Levels in 50% alcohol+TNBS group were higher than those in 50% alcohol+TNBS+ketotifen group ( P<0.05). Normal saline group and 50% alcohol group had no statistically significant difference in comparison with normal control group. In TNBS model group, serum concentration of anti-CBir1 was positively correlated with MCT concentration (r=0.648, P<0.01) and histamine concentration (r=0.751, P<0.01). CONCLUSION:The heavier degree of in-flammation in TNBS-induced colitis, the higher levels of the CBir1 and the degranulation of mast cells. There is a positive correlation between the expression of CBir1 and the degranulation of mast cells in TNBS-induced colitic mice.

11.
Immune Network ; : e18-2018.
Article in English | WPRIM | ID: wpr-715082

ABSTRACT

Mast cells integrate innate and adaptive immunity and are implicated in pathophysiological conditions, including allergy, asthma, and anaphylaxis. Cross-linking of the high-affinity IgE receptor (FcεRI) initiates diverse signal transduction pathways and induces release of proinflammatory mediators by mast cells. In this study, we demonstrated that hyperactivation of mechanistic target of rapamycin (mTOR) signaling using the mTOR activator MHY1485 suppresses FcεRI-mediated mast cell degranulation and cytokine secretion. MHY1485 treatment increased ribosomal protein S6 kinase (S6K) and eukaryotic translation initiation factor 4E-binding protein 1 (4E-BP1) phosphorylation, which are downstream targets of mTOR complex 1 (mTORC1), but decreased phosphorylation of Akt on mTOR complex 2 (mTORC2) target site serine 473. In addition, this activator decreased β-hexosaminidase, IL-6, and tumor necrosis factor α (TNF-α) release in murine bone marrow-derived mast cells (BMMCs) after FcεRI stimulation. Furthermore, MHY1485-treated BMMCs showed significantly decreased proliferation when cultured with IL-3. These findings suggested hyperactivation of mTORC1 as a therapeutic strategy for mast cell-related diseases.


Subject(s)
Adaptive Immunity , Anaphylaxis , Asthma , Cell Degranulation , Cell Proliferation , Hypersensitivity , Immunoglobulin E , Interleukin-3 , Interleukin-6 , Mast Cells , Peptide Initiation Factors , Phosphorylation , Ribosomal Protein S6 Kinases , Serine , Signal Transduction , Sirolimus , Tumor Necrosis Factor-alpha
12.
Article in English | WPRIM | ID: wpr-121497

ABSTRACT

Mast cells are major immune cells in allergy to secrete allergic mediators by a degranulation process and make and secrete inflammatory lipids and cytokines in response to antigen stimulation. An amino acid tryptophan regulates immune functions. Tryptophan ameliorates inflammatory colitis in which mast cells are engaged. However, its effects on mast cells remain to be solved. We investigated the effect of tryptophan on IgE-mediated allergic responses in the mast cells and mice. IgE-mediated passive cutaneous anaphylaxis (PCA) in mice were examined. Also IgE-mediated mast cell activation responses such as degranulation of stored granules and secretion of inflammatory lipid LTB₄ and cytokines (TNF-α and IL-4) were measured. Intraperitoneal administration of tryptophan suppressed PCA in mice. Also, in the cellular level tryptophan inhibited IgE-mediated mast cell activation such as IgE-mediated degranulation and the production of LTB₄. Also, it inhibited production of inflammatory cytokines TNF-α and IL-4. In summary, tryptophan suppressed IgE-mediated allergic activation in vivo and in vitro. Tryptophan supplementation is beneficial for IgE-mediated allergy.


Subject(s)
Animals , Colitis , Cytokines , Hypersensitivity , Immunoglobulin E , In Vitro Techniques , Interleukin-4 , Leukotriene B4 , Mast Cells , Mice , Passive Cutaneous Anaphylaxis , Tryptophan
13.
Article in Chinese | WPRIM | ID: wpr-507693

ABSTRACT

Objective To investigate the analgesic effect of warm needling and explore its relationship with the function of mast cells in the acupoint area and the specificity of acupoints.Method Sixty SD rats were randomized into blank (C), model (M), warm needling Zusanli (Z), contralateral warm needling (O), manual acupuncture (A), warm needling Futu (F), warm needling Yanglingquan(Y), warm needling Weizhong (W), disodium cromoglycate + warm needling (DSCG+Z) and saline + warm needling (Saline+Z) groups. A rat with adjuvant arthritis was used as a model of inflammatory pain (AA model). Hindpaw withdrawal latency was measured in therats during warm needling at Zusanli (ST36), Yanglingquan (SP9), Futu (ST32) and Weizhong (BL40). Pre-treatment and post-treatment rates of mast cell degranulation in the acupoint area were compared and the effect of acupoint area injection of disodium cromoglycate on the degranulation was observed by acupoint tissue section staining.Result Obvious degranulation of mast cells in the acupoint area appeared after warm needling (P>0.05). The analgesic effect of warm needling was better than that of manualacupuncture (P>0.05). After treatment, pain threshold was significantly higher in the warm needling Zusanli group than in the warm needling Futu, warm needling Yanglingquan and warm needling Weizhong groups (P>0.05). Acupuncture-induced mast cell degranulation could be prevented by disodium cromoglycate (P>0.05).Conclusion Warm needling at point Zusanli can produce an analgesic effect in adjuvant arthritis rats. Its analgesic effect is better than that of manual acupuncture. The rate of mast cell degranulation in the acupoint area is higher in the warm needling Zusanli group than in the manual acupuncture group. The analgesic effect of warm needling is reduced after pretreatment with disodium cromoglycate, a mast cell stabilizer. In an AA model, the analgesic effect of warm needling at point Zusanli is better than those of warm needling at points Yanglingquan, Futu and Weizhong.

14.
Article in English | WPRIM | ID: wpr-819420

ABSTRACT

OBJECTIVE@#To investigate the effects of friedelin (terpenoid) and 8-hydroxyisocapnolactone-2-3-diol (coumarin) with concentration 10 μM, 30 μM, and 100 μM on inhibiting mast cells (MCs) degranulation.@*METHODS@#The investigation was performed in vitro by administering each compound into rat peritoneal MCs and rat basophilic leukemia-2H3 cells followed by activation with 50 μg/mL of compound 48/80 or 1 μM of ionomycin. The concentration of histamine released from each group was measured by a high-performance liquid chromatography-fluorometry system with post-column derivatization using o-phthalaldehyde.@*RESULTS@#8-Hydroxyisocapnolactone-2-3-diol inhibited degranulation of compound 48/80 activated-rat peritoneal MCs with the histamine release percentages of 74.57%, 72.21% and 51.79% when the 10 μM, 30 μM and 100 μM concentrations were used, respectively. Where as about 81% histamine was released by the control group. Degranulation inhibition ability was also observed in ionomycin-activated rat basophilic leukemia-2H3 cells. In contrast, friedelin failed to inhibit degranulation in either cell type. The inhibition of 8-hydroxyisocapnolactone-2-3-diol was not related to the depletion of histamine synthesis as implied by the total histamine measurement.@*CONCLUSIONS@#These results exhibit the promising of 8-hydroxyisocapnolactone-2-3-diol is a potential parent structure for developing a MCs stabilizer.

15.
Anatomy & Cell Biology ; : 275-283, 2017.
Article in English | WPRIM | ID: wpr-47825

ABSTRACT

Allergic diseases are a significant health concern in developing countries. Type-A procyanidin polyphenols from cinnamon (Cinnamomum zeylanicum Blume) bark (TAPP-CZ) possesses antiasthmatic and antiallergic potential. The present study was aimed at the possible anti-allergic mechanism of TAPP-CZ against the compound 48/80 (C48/80)–induced mast cell degranulation in isolated rat peritoneal mast cells (RPMCs). TAPP-CZ (1, 3, 10, and 30 µg/ml) was incubated for 3 hours with isolated, purified RPMCs. The C48/80 (1 µg/ml) was used to induce mast cell degranulation. The mast cell viability was assessed using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay whereas histamine, β-hexosaminidase (β-HEX), and interleukin-4 (IL-4) levels were determined in RPMCs. TAPP-CZ (3, 10, and 30 µg/ml) showed significant and dose-dependent decrease in a number of degranulated cells and levels of markers (histamine, β-HEX, and IL-4) as compared with C48/80 control. In conclusion, TAPP-CZ stabilizes mast cell and cause inhibition of the allergic markers such as histamine, IL-4, and β-HEX in IgE-mediated manner. The present study supports mast cell stabilization as a possible mechanism of action of TAPP-CZ against immune respiratory disorders such as asthma and allergic rhinitis.


Subject(s)
Animals , Asthma , Cinnamomum zeylanicum , Developing Countries , Histamine , Interleukin-4 , Mast Cells , Polyphenols , Proanthocyanidins , Rats , Rhinitis, Allergic
16.
Basic & Clinical Medicine ; (12): 479-483, 2017.
Article in Chinese | WPRIM | ID: wpr-513861

ABSTRACT

Objective To explore the anti-infection mechanism of carboxyamidotriazole (CAI) through studying the effects of CAI on the proliferation, apoptosis and degranulation of RBL-2H3 mass cells.Methods Compound 48/80 (C48/80) was used to induce the model of activation and degranulation in RBL-2H3 cells.The morphological change of cell degranulation was observed by neutral red staining.The release levels of histamine and β-hexosaminidase were measured by ELISA method and chromogenic assay, respectively.The cell activity was determined by CCK-8 method.And cell apoptosis was detected by Hoechst 33342 fluorescent staining.Results Compared with the control group, 10, 20, 40 μmol/L CAI inhibited C48/80-induced degranulation of RBL-2H3 cells in different degrees.CAI (20, 40 μmol/L) reduced the histamine release (P<0.01), and CAI (40 μmol/L) decreased the β-hexosaminidase release (P<0.01).In addition, the viability and apoptosis of RBL-2H3 cells were not affected at the concentrations of CAI used.Conclusions CAI can effectively inhibit the activation and degranulation of RBL-2H3 mast cells, and this effect is not through cytotoxicity.The anti-infection effect of CAI may partially due to the down-regulation of mast cell activity.

17.
Article in Chinese | WPRIM | ID: wpr-250469

ABSTRACT

Anaphylactoid reaction (AR) is the most common adverse reaction of injection formulations, however, there are obvious drawbacks in available methods for AR detection. A novel in vitro detection method for AR was established based on fluorescent labeling and high content screen (HCS) system in present study. With the use of RBL-2H3 cells degranulation model, positive cell count was determined with specific cellular membrane fluorescent dye FM4-64 labeling vesicle recycle, and total cells count was determined with specific nucleus fluorescent dye Hochest 3334, and then the ratio of cells degranulation after drug stimulation was calculated. In order to verify the reliability of this HCS method, positive drug Compound 48/80 was first used to confirm the consistence of HCS method with the traditional β-hexosaminidase release test and the Evans blue staining ears test in mice. The results showed high consistence between HCS method and traditional testing methods, and the HCS method showed higher sensitivity than the other two tests. Then 30 samples of Danhong injection (DHI) with clinical allergy symptoms further were used to confirm the reliability of this HCS method. The HCS results showed high consistence with the clinical report, and the HCS method had the advantage in reducing the interference by drug color. Therefore, this HCS method is reliable, sensitive, simple and high-throughput method in detection of AR, applicable for the AR evaluation of injection formulations, and can provide guidance for safety of clinical application in clinical practice.

18.
Article in English | WPRIM | ID: wpr-728533

ABSTRACT

Mast cells are primary mediators of allergic inflammation. Beta-1,3-glucan (BG) protects against infection and shock by activating immune cells. Activation of the BG receptor induces an increase in intracellular Ca2+, which may induce exocytosis. However, little is known about the precise mechanisms underlying BG activation of immune cells and the possible role of mitochondria in this process. The present study examined whether BG induced mast cell degranulation, and evaluated the role of calcium transients during mast cell activation. Our investigation focused on the role of the mitochondrial calcium uniporter (MCU) in BG-induced degranulation. Black mouse (C57) bone marrow-derived mast cells were stimulated with 0.5 microg/ml BG, 100 microg/ml peptidoglycan (PGN), or 10 microM A23187 (calcium ionophore), and dynamic changes in cytosolic and mitochondrial calcium and membrane potential were monitored. BG-induced mast cell degranulation occurred in a time-dependent manner, and was significantly reduced under calcium-free conditions. Ruthenium red, a mitochondrial Ca2+ uniporter blocker, significantly reduced mast cell degranulation induced by BG, PGN, and A23187. These results suggest that the mitochondrial Ca2+ uniporter has an important regulatory role in BG-induced mast cell degranulation.


Subject(s)
Animals , Calcimycin , Calcium , Cytosol , Exocytosis , Inflammation , Ion Transport , Mast Cells , Membrane Potentials , Mice , Mitochondria , Peptidoglycan , Ruthenium Red , Shock
19.
International Eye Science ; (12): 1032-1035, 2016.
Article in Chinese | WPRIM | ID: wpr-637837

ABSTRACT

? AIM: To investigate the immunosuppression of artemisinin in the early-phase response( EPR) of allergic conjunctivitis in a murine model induced by pollen.?METHODS:Twenty-five Balb/c mice were divided into five groups at random: A: model group, B: artemisinin topical treatment group, C: oral artemisinin group, D:dexamethasone topical treatment group, E: negative control group. Balb/c mice were first sensitized with mixture of ragweed crude extract and complete Freund’s adjuvant by left footpad and root of tails injection at days 0. At days 7 and 14, mice were strengthen sensitized by intraperitoneal injection. Negative control group replacd with a mixture of equal amounts of Freund’s adjuvant and PBS mixture. Treatment time was between 21-27d:group B received 1% topical artemisinin eye drops 4 times per day; group C received artemisinin at doses of 600mg/kg orally once per day; group D received topical dexamethasone eye drops 4 times per day. The model and naive groups replaced with solvent for control. After treatment, mice were excited by ragweed pollen crude infusion drops. Within 1h after the excitation, the eyes were taken for histopathology testing, the serum for detection of specific IgE for ragweed pollen.?RESULTS:After excitation, allergy symptoms of model group were more significantly compared with negative control group and the treatment groups; allergy symptom score, mast cell degranulation ratios and the ragweed pollen specific IgE in serum in the topical treatment groups and oral group were higher than that in the negative control group, but significantly lower than that in model group.?CONCLUSION: Artemisinin topical treatment and oral treatment for mice could inhibit the symptoms of pollen allergic conjunctivitis, early-phase response of mast cell degranulation ratio and specific IgE generation for ragweed pollen, suggesting that artemisinin has some therapeutic effect for pollen allergic conjunctivitis.

20.
Chinese Journal of Immunology ; (12): 1153-1156, 2016.
Article in Chinese | WPRIM | ID: wpr-495094

ABSTRACT

Objective:To investigate the effect of Luteolin on the degranulation function of RBL-2H3 mast cells and its mechanism of action.Methods:DNP-BSA-IgE was used to establish the sensitized cell model in RBL-2H3 cells.Cell viability was exa mined by MTT assay at different concentrations (5,15,25 μmol/L) of Luteolin pretreated RBL-2H3 cells.The effect of Luteolin on β-hexosa minidase(β-hex) and TNF-αwas evaluated by ELISA.The change of Ca2+influx was detected by Flou-4AM calcium ion fluorescent probe.The expression of AKT and P-AKT were detected by Western blot.Results:The sensitized cell model was established successfully .Cell viability had not significantly changes stimulated by different concentrations of Luteolin ,and a significantly inhibition role showed the release of β-hex and TNF-αfrom RBL-2H3 cells.The concentration of intracellular Ca 2+ was significantly decreased.However,the Western blot results showed that the level of Phosphorylation AKT was decreased .Conclusion: Luteolin suppress RBL-2H3 cell degranulation in dose dependence ,which may via impacting the Ca2+influx and the activity of AKT.

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