1.The effect of Qi'ao Decoction on ovalbumin induced and lipopolysaccharide enhanced severe asthma mice and its mechanism.
Chen-Xue JIANG ; Xin-Sheng FAN ; Peng-Cheng GU ; Hui-Qin XU
Chinese Journal of Natural Medicines (English Ed.) 2013;11(6):638-644
AIM:
To evaluate the effect of Qi'ao Deocoction (QAD) on the inflammation and hyperresponsiveness of asthma mice.
METHODS:
120 Balb/C mice were randomly divided into six groups: normal group, model group, dexamethasone group, high dose QAD group, medium dose QAD group and low dose QAD group. The asthma model was reproduced in Balb/C mice sensitized by ovalbumin, challenged by OVA and LPS. The mice of the normal group were sensitized, challenged and intranasally instilled by PBS. On day 28-34, 6.7, 13.4 and 26.8 g · kg(-1) Qi'ao Decoction were administrated; 0.002 4 g · kg(-1) dexamethasone solution was given to the dexamethasone group; normal and model groups were given the same amount of normal saline. Bronchoalveolar lavage fluid, airway hyperresponsiveness, lung histopathology and cytokines were then collected and analyzed.
RESULTS:
Compared with normal group, total cellular score, the number of macrophages, lymphocytes, eosinophils and neutrophils of model group significantly increased (P < 0.01). Compared with model group, the administration of dexamethasone induced a significant decrease in eosinophils and neutrophils (P < 0.05, P < 0.01). The number of eosinophils, which plays an important role in airway inflammatory reaction of asthma, of the three QAD groups all decreased (P < 0.01). RL before and after Ach (5 mg · mL(-1)) stimulation in the model group both overtook that in the normal group (P < 0.01). Compared with model group, dexamethasone group, high dose QAD group, medium dose QAD group and low dose QAD group groups all had significantly lower RL before and after Ach stimulation (P < 0.01). Normal pulmonary histopathology was found in the normal group. In the model group, mice exhibited marked increases in inflammatory cell infiltration, mostly including neutrophils and macrophages, perivascular inflammation and thickened alveolus wall (P < 0.01). Dexamethasone application mitigated inflammation around the bronchi (P < 0.05). These histopathological changes were ameliorated in the three decoction groups (P < 0.01, P < 0.05). In addition, alveolus and airway wall lesions of medium dose QAD group and high dose QAD group were reduced, the number of inflammatory cells infiltrated around the walls decreased, no clear degeneration of bronchial epithelial cells was found, and exudates in bronchi declined in different degrees. Compared with normal group, IFN-γ and IL-12 of model group significantly decreased, while IL-4 increased, showing statistic difference (P < 0.05). Compared with model group, IFN-γ and IL-12 level of dexamethasone group went up too, but IL-4 declined (P < 0.05). The level of IFN-γ of medium dose QAD group and high dose QAD group both increased; IL-4 and IL-12 of medium dose group were found significant differences (P < 0.05); but none of the cytokines of low dose QAD group showed statistical significance (P > 0.05).
CONCLUSION
QAD can significantly inhibit airway inflammation and airway hyperresponsiveness of mice with severe asthma induced by ovalumin and lipopolysaccharide, adjust the balance of cytokines, and improve lung histopathological condition. So, it exhibits great effect on severe asthma.
Animals
;
Asthma
;
chemically induced
;
drug therapy
;
immunology
;
pathology
;
Drugs, Chinese Herbal
;
administration & dosage
;
Female
;
Humans
;
Interleukin-12
;
immunology
;
Interleukin-4
;
immunology
;
Lipopolysaccharides
;
adverse effects
;
immunology
;
Lung
;
immunology
;
pathology
;
Mice
;
Mice, Inbred BALB C
;
Ovalbumin
;
adverse effects
;
immunology
2.Acute exposure to silica nanoparticles aggravate airway inflammation: different effects according to surface characteristics.
Hye Jung PARK ; Jung Ho SOHN ; Yoon Ju KIM ; Yoon Hee PARK ; Heejae HAN ; Kyung Hee PARK ; Kangtaek LEE ; Hoon CHOI ; Kiju UM ; In Hong CHOI ; Jung Won PARK ; Jae Hyun LEE
Experimental & Molecular Medicine 2015;47(7):e173-
Silica nanoparticles (SNPs) are widely used in many scientific and industrial fields despite the lack of proper evaluation of their potential toxicity. This study examined the effects of acute exposure to SNPs, either alone or in conjunction with ovalbumin (OVA), by studying the respiratory systems in exposed mouse models. Three types of SNPs were used: spherical SNPs (S-SNPs), mesoporous SNPs (M-SNPs), and PEGylated SNPs (P-SNPs). In the acute SNP exposure model performed, 6-week-old BALB/c female mice were intranasally inoculated with SNPs for 3 consecutive days. In the OVA/SNPs asthma model, the mice were sensitized two times via the peritoneal route with OVA. Additionally, the mice endured OVA with or without SNP challenges intranasally. Acute SNP exposure induced significant airway inflammation and airway hyper-responsiveness, particularly in the S-SNP group. In OVA/SNPs asthma models, OVA with SNP-treated group showed significant airway inflammation, more than those treated with only OVA and without SNPs. In these models, the P-SNP group induced lower levels of inflammation on airways than both the S-SNP or M-SNP groups. Interleukin (IL)-5, IL-13, IL-1beta and interferon-gamma levels correlated with airway inflammation in the tested models, without statistical significance. In the mouse models studied, increased airway inflammation was associated with acute SNPs exposure, whether exposed solely to SNPs or SNPs in conjunction with OVA. P-SNPs appear to be relatively safer for clinical use than S-SNPs and M-SNPs, as determined by lower observed toxicity and airway system inflammation.
Animals
;
Asthma/*chemically induced/pathology
;
Female
;
Inflammation/*chemically induced/pathology
;
Interferon-gamma/analysis
;
Interleukins/analysis
;
Lung/drug effects/*pathology
;
Mice, Inbred BALB C
;
Nanoparticles/*adverse effects/chemistry
;
Ovalbumin/adverse effects
;
Polyethylene Glycols/adverse effects/chemistry
;
Silicon Dioxide/*adverse effects/chemistry
;
Surface Properties
3.Mechanism of "Ephedrae Herba-Descurainiae Semen Lepidii Semen" combination in treatment of bronchial asthma based on network pharmacology and experimental verification.
Bei-Bei ZHANG ; Meng-Nan ZENG ; Qin-Qin ZHANG ; Ru WANG ; Ju-Fang JIA ; Peng-Li GUO ; Meng LIU ; Wei-Sheng FENG ; Xiao-Ke ZHENG
China Journal of Chinese Materia Medica 2022;47(18):4996-5007
This study aims to investigate mechanism of "Ephedrae Herba-Descurainiae Semen Lepidii Semen" combination(MT) in the treatment of bronchial asthma based on network pharmacology and in vivo experiment, which is expected to lay a theoretical basis for clinical application of the combination. First, the potential targets of MT in the treatment of bronchial asthma were predicted based on network pharmacology, and the "Chinese medicine-active component-target-pathway-disease" network was constructed, followed by Gene Oncology(GO) term enrichment and Kyoto Encyclopedia of Genes and Genomes(KEGG) pathway enrichment of the potential targets. Molecular docking was used to determine the binding activity of key candidate active components to hub genes. Ovalbumin(OVA, intraperitoneal injection for sensitization and nebulization for excitation) was used to induce bronchial asthma in rats. Rats were classified into control group(CON), model group(M), dexamethasone group(DEX, 0.075 mg·kg~(-1)), and MT(1∶1.5) group. Hematoxylin and eosin(HE), Masson, and periodic acid-Schiff(PAS) staining were performed to observe the effect of MT on pathological changes of lungs and trachea and goblet cell proliferation in asthma rats. The levels of transforming growth factor(TGF)-β1, interleukin(IL)6, and IL10 in rat serum were detected by enzyme-linked immunosorbent assay(ELISA), and the mRNA and protein levels of mitogen-activated protein kinase 8(MAPK8), cyclin D1(CCND1), IL6, epidermal growth factor receptor(EGFR), phosphatidylinositol 3-kinase(PI3 K), and protein kinase B(Akt) by qRT-PCR and Western blot. Network pharmacology predicted that MAPK8, CCND1, IL6, and EGFR were the potential targets of MT in the treatment of asthma, which may be related to PI3 K/Akt signaling pathway. Quercetin and β-sitosterol in MT acted on a lot of targets related to asthma, and molecular docking results showed that quercetin and β-sitosterol had strong binding activity to MAPK, PI3 K, and Akt. In vivo experiment showed that MT could effectively alleviate the symptoms of OVA-induced asthma rats, improve the pathological changes of lung tissue, reduce the production of goblet cells, inhibit the inflammatory response of asthma rats, suppress the expression of MAPK8, CCND1, IL6, and EGFR, and regulate the PI3 K/Akt signaling pathway. Therefore, MT may relieve the symptoms and inhibit inflammation of asthma rats by regulating the PI3 K/Akt signaling pathway, and quercetin and β-sitosterol are the candidate active components.
Animals
;
Asthma/drug therapy*
;
Cyclin D1
;
Dexamethasone/adverse effects*
;
Drug Combinations
;
Drugs, Chinese Herbal/therapeutic use*
;
Eosine Yellowish-(YS)/adverse effects*
;
Ephedra
;
ErbB Receptors
;
Hematoxylin/therapeutic use*
;
Interleukin-10
;
Interleukin-6
;
Mitogen-Activated Protein Kinase 8/therapeutic use*
;
Molecular Docking Simulation
;
Network Pharmacology
;
Ovalbumin/adverse effects*
;
Periodic Acid/adverse effects*
;
Phosphatidylinositol 3-Kinases
;
Proto-Oncogene Proteins c-akt/metabolism*
;
Quercetin
;
RNA, Messenger
;
Rats
4.Effect of Yupingfeng San against OVA-induced allergic asthma in mice.
Hui-Zhu WANG ; Min HONG ; Li-Li GUI ; Yong-Qing HUA ; Hui-Qin XU
China Journal of Chinese Materia Medica 2013;38(7):1052-1055
OBJECTIVETo observe the effect of Yupingfeng San (YPFS) against OVA-induced allergic asthma in mice.
METHODMice were injected with OVA to establish the allergic asthma model. They were abdominally injected with 20 microg OVA on day 0 and 14, and inhaled aerosol 0.5% OVA solution for 20 min for seven days. The blank control group was administrated with equal volume of saline. YPFS groups with different doses were administrated intragastrically with YPFS every day, with the crude drug dosage of 3.25, 6.5, 13 g x kg(-1), respectively. The model group and control group were administrated with equal volume of saline. The positive control group was given intraperitoneally injected with 1 mg x kg(-1) DEX since aerosol inhalation. Blood was drawn after the last OVA aerosol inhalation to count the number of Eosnophils (Eos) in blood and detect IgE in serum; BALF was collected to count the number of cells and classify; right lung tissues were evenly grinded to detect cytokines IL-4 and IFN-gamma, and left upper lung lobes were collected for pathologic histology.
RESULTThe level of Eos and IgE in serum increased significantly in the model group, and a large number of Eos were detected in BALF. Histopathological changes in lung showed bronchial serous exudation, tubular epithelial cells exfoliation, tube narrowing, widened alveolar septum, and bronchial periarterial lymphocytes infiltration. Homogenate of lung tissues showed increase of IL-4, and decrease in IFN-gamma/IL-4 ratio. YPFS groups with different doses displayed decrease of Eos in blood and BALF and IgE content in serum, and relief of pathologic changes in above models. Meanwhile, IL-4 content in homogenate of lung tissues decreased, with the increase in IFN-gamma/IL-4 ratio.
CONCLUSIONYPFS shows the inhibitory effects on OVA-induced allergic asthma, involving down regulation of Eos and IgE levels in blood of asthma mice, and infiltration of inflammatory cells in lung tissues. Meanwhile, it can reduce IL-4 in lung homogenates, increase IFN-gamma/IL-4, and inhibits Th2 polarization.
Animals ; Asthma ; chemically induced ; drug therapy ; immunology ; Disease Models, Animal ; Drugs, Chinese Herbal ; administration & dosage ; Eosinophils ; drug effects ; immunology ; Humans ; Interferon-gamma ; immunology ; Interleukin-4 ; immunology ; Male ; Mice ; Mice, Inbred BALB C ; Ovalbumin ; adverse effects
5.Effects of fulvotomentoside on inflammatory factors and antiinflammatory factors in intestine of ovalbumin-sensitized BALB/c mice.
Chinese Journal of Pediatrics 2010;48(7):520-525
OBJECTIVETo investigate the effects of fulvotomentoside (Ful) on inflammatory factors and antiinflammatory factors in intestine of ovalbumin (OVA)-sensitized BALB/c mice, and to explore the mechanisms of its anti-food allergy effect.
METHODTwenty-four female BALB/c mice aged 6 weeks fed with ovalbumin-free feed were randomly divided into 3 groups, food allergy (FA) group, Ful group and normal saline (NS) group. Mice in FA and Ful groups were sensitized intraperitoneally two times with OVA and challenged intragastrically with OVA. Mice in Ful group were treated with 200 mg/kg of Ful by subcutaneous injection once daily for 22 days. The mice in FA and NS groups were used as positive control and negative control, respectively, and were treated with normal saline solution by subcutaneous injection for 22 days. Just 48 hours after the last challenge, the mice in each group were sacrificed and specimens of jejunum were taken. The mRNA expressions of transforming growth factor β1 (TGF-β1), interleukin-6 (IL-6), interleukin-17A (IL-17A) and forkhead box P3 (Foxp3) in jejunum were detected by reverse transcription-PCR (RT-PCR). The protein expressions of TGF-β1, IL-6, and IL-17A in jejunum were detected by immunohistochemical method. The activation of neutrophils in jejunum was assayed by the levels of MPO.
RESULTThe expressions of TGF-β1, IL-6, IL-17A mRNA [(0.370 ± 0.013), (0.475 ± 0.015), (0.541 ± 0.013)] and related protein [(53,075.70 ± 20,727.06), (256,881.66 ± 36,561.79), (435,064.25 ± 69,911.48)] in jejunum were increased and the Foxp3 mRNA [(0.231 ± 0.014) vs. (0.365 ± 0.015)] expression was decreased in group FA. After the treatment with Ful, IL-6 and IL-17A mRNA [(0.196 ± 0.005), (0.204 ± 0.008)] and protein [(114,040.30 ± 20,295.25), (218,200.74 ± 30,077.69)] expressions were decreased and Foxp3 mRNA (0.578 ± 0.021) expression was increased, and no change of TGF-β1 expression was unchanged. There were no significant differences of the levels of MPO among the three groups.
CONCLUSIONInflammatory reaction which was characterized by the increase of IL-6 and IL-17A expressions was found in intestine of ovalbumin-sensitized BALB/c mice. Ful could decrease overexpression of IL-6 and IL-17A, and increase the expression of specific transcription factor Foxp3 of regulatory T cells significantly in intestine. It may be one of the mechanisms that Ful improved intestinal inflammation.
Animals ; Female ; Food Hypersensitivity ; metabolism ; Forkhead Transcription Factors ; metabolism ; Inflammation ; Interleukin-17 ; metabolism ; Interleukin-6 ; metabolism ; Intestines ; drug effects ; metabolism ; Mice ; Mice, Inbred BALB C ; Oleanolic Acid ; analogs & derivatives ; pharmacology ; Ovalbumin ; adverse effects ; Saponins ; pharmacology ; Transforming Growth Factor beta1 ; metabolism
6.Allergic airway response associated with the intestinal microflora disruption induced by antibiotic therapy.
Chong-hai LIU ; Xi-qiang YANG ; Chun-hua LIU ; Yun HE ; Li-jia WANG
Chinese Journal of Pediatrics 2007;45(6):450-454
OBJECTIVEOver the past several decades, there has been a significant increase in allergy and asthma in the world, which correlates with alterations in microflora and widespread use of antibiotics. The authors have developed a mouse model of antibiotics-induced microbiota disruption. In that model, mice were challenged by intranasal exposure to Aspergillus fumigatus allergens to explore the relation of allergic airway response and intestinal microflora disruption.
METHODSSixty female BALB/c mice were divided at random into 6 groups with 10 mice in each. (1) First antibiotic therapy group: the mice were given oral cefoperazone for 7 days, on day 7, mice were inoculated with Candida albicans (10(9)/ml, 50 microl) orally. (2) First control group: the mice were treated as first antibiotic therapy group, but cefoperazone and Candida albicans were replaced by saline. The mice in groups (1) and (2) were sacrificed on day 8, and cecal contents were collected for quantitative analysis of the intestinal bacterial flora. (3) Antibiotic therapy and challenge group: the mice were treated as the first antibiotic therapy group, then challenged (day 9 and 16) by intranasal exposure to Aspergillus fumigatus allergen. (4) Second antibiotic therapy group: the mice were treated as the first antibiotic therapy group, then challenged (day 9 and 16) by intranasal exposure to saline. (5) Challenge group: the mice were treated as the first control group, then challenged (day 9 and 16) by intranasal exposure to Aspergillus fumigatus allergen. (6) Second control group: the mice were treated as the first control group, then challenged (day 9 and 16) by intranasal exposure to saline. The mice in (3) - (6) group were killed for analysis of allergic airway response on day 19.
RESULTSThe quantity of Enterobacteriaceae, Enterococcus, Bifidobacterium and Lactobacillus in first antibiotic therapy group was significantly lower than that in the first control group, the quantity of Candida albicans increased in the first antibiotic therapy group as compared with the first control group. Mice intestinal microflora were disrupted with weight reduction and increased moisture in feces. After challenging with Aspergillus fumigatus allergens via intranasal inhalation, the total cell count, eosinophils, lymphocytes and neutrophils increased in BALF, especially in bronchoalveolar lavage fluid (BALF) from the mice in antibiotic therapy and challenge groups. IL-4 level in BALF from antibiotic therapy and challenge group (45.35 +/- 2.36) pg/ml was higher than that in the second control group (35.32 +/- 2.53) pg/ml. The expression of GATA-3 mRNA in the mice lung tissue (0.569 +/- 0.023) was higher than that in the second control group (0.410 +/- 0.020), and the ratios of T-bet/GATA-3 (0.578 +/- 0.021) decreased as compared with that in the second control group (0.804 +/- 0.035). IFN-gamma level in BALF from any group was not significantly different. In the absence of antibiotics, mice exposed to Aspergillus fumigatus allergen did not develop an allergic response in the airways.
CONCLUSIONSThe allergic (Th2) immune response can be induced by airway challenge with Aspergillus fumigatus allergen in the mice in which the intestinal microflora disruption resulted from antibiotic therapy, this result suggests that the intestinal microflora disruption resulted from antibiotic therapy is a risk factor for allergy and asthma.
Animals ; Anti-Bacterial Agents ; adverse effects ; Antibiosis ; Aspergillus fumigatus ; chemistry ; growth & development ; Asthma ; drug therapy ; microbiology ; Bronchoalveolar Lavage Fluid ; microbiology ; Cefoperazone ; therapeutic use ; Disease Models, Animal ; Eosinophils ; drug effects ; microbiology ; Female ; Hypersensitivity ; drug therapy ; microbiology ; Hypersensitivity, Immediate ; microbiology ; Intestines ; drug effects ; microbiology ; physiopathology ; Lung ; drug effects ; microbiology ; Mice ; Mice, Inbred BALB C ; Ovalbumin ; adverse effects ; immunology ; Respiratory System ; microbiology
7.Therapeutic effects of tolerogenic dendritic cells on ovalbumin allergic BALB/c mice.
Li WANG ; Yan HU ; Qian CHENG
Chinese Journal of Pediatrics 2010;48(11):839-842
OBJECTIVETo investigate the therapeutic effects of AdCTLA4 Ig/Adα4β7-dendritic cell (DC) in the ovalbumin (OVA) allergic BALB/c mice.
METHODTwenty-four Female BALB/c mice aged 4-6 weeks fed on the ovalbumin-free feed were randomly divided into 3 groups: (1) treatment group: the OVA sensitized mice were injected with tolerogenic AdCTLA4-Ig/Adα4β7-DC after being challenged with OVA. (2) negative control group: mice were sensitized and challenged with normal saline. (3) positive control group: mice were sensitized and challenged with OVA. The level of the OVA-specific IgE in serum was measured by ELISA. The jejunal samples were observed histologically after HE staining. The level of IL-10 and TGF-β in murine intestine was detected by immunohistochemical and immunofluorescent methods, respectively.
RESULTCompared with the positive group, caudal vein injection with AdCTLA4Ig/Adα4β7-DC in treatment group reduced the level of the OVA-specific IgE in the serum remarkably, inhibited the inflammatory reactions of the intestine and increased the expression of the IL-10 in intestine significantly. There was no significant change in the expression of TGF-β in each group.
CONCLUSIONThis study demonstrated that caudal vein injection with tolerogenic AdTLA4Ig/Adα4β7-DC may be of potential research value in the treatment of food allergy by inducing immune tolerance in vivo.
Animals ; Dendritic Cells ; immunology ; Disease Models, Animal ; Egg Hypersensitivity ; therapy ; Female ; Immune Tolerance ; Immunoglobulin E ; blood ; Inflammation ; Interleukin-10 ; immunology ; Mice ; Mice, Inbred BALB C ; Ovalbumin ; adverse effects ; Transforming Growth Factor beta ; immunology
8.Effect of food allergy early in life on visceral hyperalgesia in the developing rats and role of substance P abnormal expression in colon.
Bin WU ; Qian-kun LAI ; Jun-hong LIU ; Chun LIN
Chinese Journal of Pediatrics 2013;51(3):172-177
OBJECTIVETo explore the effect of food allergy (FA) on the development of visceralgia sensibility and the substance P (SP) expression in colon of developing rats with FA.
METHODThree-week old female Sprague-Dawley (SD) rats were randomly divided into two groups (n = 10 in each). The rats in FA group were sensitized with ovalbumin (OVA) 40 µg and Al(OH)3 1 mg suspension solution (0.2 ml) intraperitoneal (i.p.) injection on day 0, only OVA 40 µg solution i.p. on day 2, 4, 7, 9, 11, respectively, and the rats were challenged by gavage with OVA solution 30 mg (2 ml) on day 20, 24, 28, 30. The rats in non-sensitized (NS) group were not challenged except handled in the same ways. The serum OVA-IgE were determined by enzyme-linked immuno sorbent assay (ELISA) on day 0, 30. Jejunum segments were used to observe morphological structure, the expression of eosinophils, and the density and the percentage of degranulation of mast cells (MC). The rats were appraised for the pain sensibility of intestinal tract under colorectal distension irritation by the electrophysiological method on external oblique in the 18-24 hr after the last challenge. Colons were used to analyze the expression of SP through immunohistochemical staining and computer image analyzing system.
RESULTThe serum OVA-IgE concentration and the eosinophils, mast cell, the percentage of mast cells degranulation in FA group were more than NS group (P < 0.01). The amplitudes of spike external oblique muscle of abdomen (EOMA, µV) of the FA group under the colorectal distension (CRD) pressures at 0, 15, 30, 45, 60, 75 mm Hg were (17.74 ± 0.72), (18.63 ± 1.72), (22.55 ± 1.70), (28.63 ± 7.00), (33.97 ± 7.34), (37.26 ± 8.40), and (17.43 ± 1.18), (17.27 ± 1.16), (17.73 ± 1.42), (19.55 ± 3.54), (23.29 ± 5.46), (25.20 ± 4.75) in NS group. With the CRD pressure increased, the amplitudes of spike EOMA increased significantly. There were significant differences between groups under the CRD pressures at 30, 45, 60, 75 mm Hg (F = 47.470, 13.367, 13.317, 15.390, P < 0.01). The expressions of colons SP in FA group and NS group are 247.12 ± 90.83 and 103.90 ± 58.94, respectively (t = 4.183, P < 0.01).
CONCLUSIONSensitization through i.p. pathway and challenge by gavage with OVA in early life could result in FA in young SD rats. FA in early life enabled the amplitudes of spike EOMA and the expression of colons SP increase significantly. It may be related to increase in amount and degranulation of MC and SP abnormal expression in colon, which could lead to the development of visceralgia sensibility.
Animals ; Colonic Diseases, Functional ; metabolism ; Disease Models, Animal ; Electrophysiology ; Female ; Food Hypersensitivity ; complications ; metabolism ; Hyperalgesia ; etiology ; metabolism ; physiopathology ; Intestinal Mucosa ; metabolism ; pathology ; Mast Cells ; metabolism ; Ovalbumin ; adverse effects ; Pain Threshold ; Rats ; Rats, Sprague-Dawley ; Stress, Psychological ; Substance P ; metabolism
9.Effect of Maxing Shigan Decoction and dissembled prescriptions against airway inflammation in RSV-aggravated asthma and mechanism of regulating TRPV1.
Meng-Wen LI ; Xin-Sheng FAN ; Li-Ping ZHOU ; Mo LIU ; Er-Xin SHANG
China Journal of Chinese Materia Medica 2022;47(21):5872-5881
This study investigated the effect of Maxing Shigan Decoction(MXSGD) and its disassembled prescriptions against the airway inflammation in respiratory syncytial virus(RSV)-aggravated asthma and the regulation of transient receptor potential vanilloid-1(TRPV1). To be specific, ovalbumin(OVA) and RSV were used to induce aggravated asthma in mice(female, C57BL/6). Then the model mice were intervened by MXSGD and the disassembled prescriptions. The eosinophil(EOS) in peripheral blood, inflammatory cells in bronchoalveolar lavage fluid(BALF), enhanced pause(Penh) variation, and lung pathological damage in each group were observed, and the changes of interleukin(IL)-4, IL-13, substance P(SP), and prostaglandin E2(PGE2) in BALF were mea-sured by enzyme-linked immunosorbent assay(ELISA). Quantitative real time polymerase chain reaction(qPCR) and Western blot were used to detect mRNA and protein of TRPV1 in mouse lung tissue. In the in vitro experiment, 16 HBE cells were stimulated with IL-4 and RSV. Then the changes of TRPV1 expression after the intervention with the serum containing MXSGD and its disassembled prescriptions were observed. Besides, the intracellular Ca~(2+) level after the stimulation with TRPV1 agonist was evaluated. The results showed that the mice in the model group had obvious asthma phenotype, the levels of various inflammatory cells in the peripheral blood and BALF and Penh were significantly increased(P<0.05, P<0.01), and the lung tissue was severely damaged compared with the control group. Compared with the model group, the levels of EOS in the peripheral blood and BALF were significantly decreased in the MXSGD group, the SG group and the MXC group(P<0.05, P<0.01). The levels of WBC and neutrophils in BALF were significantly decreased in the MXSGD group and SG group(P<0.01), the levels of neutrophils in BALF were decreased in the MXC group(P<0.05). The improvement effect of the MXGSD on the level of inflammatory cells in peripheral blood and BALF was better than that of two disassembled groups(P<0.05, P<0.01). After 50 mg·mL~(-1) acetylcholine chloride stimulation, the Penh values of the MXSGD group and the MXC group significantly decreased(P<0.01), and the Penh value of the SG group decreased(P<0.05). The levels of IL-4, IL-13, PGE2 and SP in BALF could be significantly decreased in the MXSGD group(P<0.05, P<0.01), the levels of IL-13 and PGE2 in BALF could be decreased in the MXC group(P<0.05, P<0.01), and the levels of IL-13, PGE2 and SP in BALF could be decreased in the SG group(P<0.05, P<0.01). MXSGD could down-regulate the protein and mRNA expression of TRPV1 in lung tissue(P<0.05, P<0.01). The serum containing MXSGD and its disassembled prescriptions could down-regulate TRPV1 expression in 16 HBE cells stimulated by IL-4 combined with RSV and inhibit the inward flow of Ca~(2+) induced by TRPV1 agonist, especially the serum containing MXSGD which showed better effect than the serum containing disassembled ones(P<0.05). In vivo and in vitro experiments verified the protective effect of MXSGD and its disassembled prescriptions against airway inflammation in RSV-exacerbated asthma, the whole decoction thus possessed synergy in treating asthma, with better performance than the dissembled prescriptions. Different groups of prescription had made contributions in improving airway hyperresponsiveness, anti-allergy and anti-inflammation. The mechanism is the likelihood that it regulates TRPV1 channel and levels of related inflammatory mediators.
Female
;
Mice
;
Animals
;
Interleukin-13/metabolism*
;
Interleukin-4/metabolism*
;
Dinoprostone
;
Mice, Inbred BALB C
;
Mice, Inbred C57BL
;
Asthma/chemically induced*
;
Lung
;
Bronchoalveolar Lavage Fluid
;
Ovalbumin/adverse effects*
;
Inflammation/metabolism*
;
RNA, Messenger/metabolism*
;
Prescriptions
;
Disease Models, Animal
;
TRPV Cation Channels/metabolism*
10.The expression and significance of NF-kappaB in the rat model of otitis media with effusion experimentally induced by ovalbumin.
Shouqin ZHAO ; Hua LIU ; Demin HAN ; Yin XIA ; Erzhong FAN ; Yang WANG ; Jie LI ; Yuanyuan REN
Journal of Clinical Otorhinolaryngology Head and Neck Surgery 2008;22(16):751-755
OBJECTIVE:
To investigate the T helper cells (Th) predominant differentiation and the modulation of nuclear transcription factors kappa B (NF-kappaB) in middle ear of rat model of otitis media with effusion (OME).
METHOD:
Sixteen SD rats were randomly divided into OME (Exp group) and control group (Con group). The expression of NF-kappaB were observed by immunohistochemistry. The level of IFN-gamma and IL-4 in tympanic lavage fluid (TLF) were determined by ELISA.
RESULT:
As compared to the Con group , the level of IL-4 and the ratio of Th2/Th1 (IL-4/IFN-gamma) in TLF of Exp group significantly increased (P<0.05), when no significant difference in IFN-gamma levels in TLF was found. The ratio of NF-kappaB p65 positive cells to white cells in temporal bone marrow smears and middle ear mucosa of Exp group was significantly higher than that of Con group (P<0.05). The expression of NF-kappaB p65 in temporal bone marrow smears and middle ear mucosa was signficantly positively correlated with the concentration of IL-4 in TLF of Exp group (P<0.05).
CONCLUSION
The middle ear is capable of mounting an allergic response and subsequent formation of effusion. There is Thl/Th2 immune response imbalance, which polarizes toward Th2 response in the middle ear microenvironment of allergic OME rat model. Moreover , NF-kappaB may participate in regulating Th2 predominant reaction.
Animals
;
Interleukin-4
;
metabolism
;
Male
;
NF-kappa B
;
metabolism
;
Otitis Media with Effusion
;
chemically induced
;
immunology
;
metabolism
;
Ovalbumin
;
adverse effects
;
Rats
;
Rats, Sprague-Dawley
;
Th1 Cells
;
immunology
;
Th2 Cells
;
immunology