Objective: To investigate the effects of Yanghe Pingchuan Granules on airway remodeling in asthmatic rats, and to explore the mechanism of Interleukin-6/Janus kinase 2/ Signal transducing activator of transcription 3(...Objective: To investigate the effects of Yanghe Pingchuan Granules on airway remodeling in asthmatic rats, and to explore the mechanism of Interleukin-6/Janus kinase 2/ Signal transducing activator of transcription 3(IL-6/JAK2/STAT3) signal axis. Methods: We separated 42 healthy male SD rats into two groups, a control group (7) and a model group (35).The model group was sensitized with a combination of ovalbumin (OVA) and aluminum hydroxide for 2 weeks, while the control group was given an equal amount of physiological saline.After 2 weeks, the modeling group was randomly divided into Model group, Yanghe Pingchuan Granules high, medium and low dose groups and Dexamethasone group, each group consisted of 7 animals. After 4 weeks, OVA atomization and gavage were used for stimulation and treatment. Yanghe Pingchuan Granules high, middle and low groups were given 15.48, 7.74, 3.87 g∙kg-1 Yanghe Pingchuan Granules daily, dexamethasone group was given 0.0625 mg∙kg-1 dexamethasone daily, and the other groups were given the same amount of normal saline. HE, PAS and Masson staining were used to observe the lung histopathological changes in rats. The levels of interleukin-6, IL-23 and IL-17A were detected by ELISA. The expression levels of JAK-2, P-JAK2, STAT3 and P-STAT3 in lung tissues were detected by Western blot. Real-time quantitative polymerase chain reaction (qRT-PCR) was used to detect the mRNA expression levels of IL-6, JAK2 and STAT3 in rat lung tissue. Results: The lung tissue structure of the model group was severely damaged compared to the control group, accompanied by a great many of inflammatory cell infiltration, goblet cell hyperplasia, subepithelial collagen fiber deposition and airway epithelial thickening were more obvious. The expressions of IL-6, IL- 23 and IL-17A in serum were significantly increased (P<0.01), the protein expression levels of JAK-2, P-JAK2, STAT3 and P-STAT3 and the mRNA expression levels of IL-6, JAK2 and STAT3 in lung tissue were significantly increased (P<0.01);Compared with the model group, inflammatory cell infiltration, goblet cell proliferation, subepithelial collagen fiber deposition and airway epithelial thickening were significantly reduced in each administration group, and the expressions of IL-6, IL-23 and IL-17A in serum were significantly decreased (P< 0.01). The protein expression levels of JAK-2, P-JAK2, STAT3 and P-STAT3 and mRNA expression levels of IL-6, JAK2 and STAT3 in lung tissue were significantly decreased (P<0.01). Conclusion: Yanghe Pingchuan Granules can significantly alleviate airway remodeling in asthmatic rats, and its mechanism may be through inhibiting the IL-6/JAK2/STAT3 signal axis.展开更多
Objective:To establish and verify aβ_(2)-AR desensitization asthma mice model.Methods:A total of 30 SPF male BALB/c mice were randomly divided into blank group,the common asthma group,andβ_(2)-AR desensitization ast...Objective:To establish and verify aβ_(2)-AR desensitization asthma mice model.Methods:A total of 30 SPF male BALB/c mice were randomly divided into blank group,the common asthma group,andβ_(2)-AR desensitization asthma model group.Asthma model was established,and on this basis,the method of atom-izing inhalation and intraperitoneal injections of salbutamol was used to prepareβ_(2)-AR desensitization asthma model.After the last stimulation on the 21st day of modeling,the airway resistance of mice was measured.ELISA was used to detect the content of serum IgE;HE staining was used to observe the lung organization degree of infla-mmatory cell infiltration;Western blot method was used to detect theβ_(2)-AR content in lung tissue,RT-PCR was used to detect theβ_(2)-ARmRNA expressionin lung tissue.Results:Compared with the blank group,as acetyl choline(Mch)levels increased,groups of OVA induced airway resistance increases;but theβ_(2)-AR desensitization asthma model group increased airway resistance was more significant(P<0.05);compared with the blank group,IgE levels of common asthma group andβ_(2)-AR desensitization asthma model group elevated(P<0.01).The pathological histology observation found theβ_(2)-AR desensitization asthma airway inflammation infiltration in mice,the excessive mucus secretion and collagen deposition,and the pathological performance obviously increase compared with the common asthma group;β_(2)-AR content in the lung tissue ofβ_(2)-AR desensitization asthma model in mice,β_(2)-AR mRNA expression level in the blank group and common asthma model group were significantly decreased(P<0.05).Conclusion:Theβ_(2)-AR desensitization asthma mouse model was successfully established,and the buildingcycle was short.展开更多
Objective:To evaluate the therapeutic effect of Glycyrrhetinic Acid on cough variant asthma(CVA)mice and to investigate the possible mechanism in reducing lung inflammation.Methods:48 young female Balb/c mice were div...Objective:To evaluate the therapeutic effect of Glycyrrhetinic Acid on cough variant asthma(CVA)mice and to investigate the possible mechanism in reducing lung inflammation.Methods:48 young female Balb/c mice were divided into Control,CVA,Prednisone Acetate,Glycyrrhetinic Acid high-dose,Glycyrrhetinic Acid middle-dose and Glycyrrhetinic Acid lowdose groups randomly,with 8 mice in each group.The CVA mice model was established by ovalbumin(OVA)sensitization and OVA challenge,the animal asthma behavior was observed after drug administration,and the index of the lung of mice were recorded.The level of OVAsIgE in the bronchoalveolar lavage fluid(BALF)was tested by ELISA.The pathological changes of the lung tissue were observed by Hematoxylin and Eosin(H&E)staining.The protein expressions of T-bet,IFN-γ,Gata3,IL-4 and IL-13 in the lung tissue were determined by Western blot.Results:Compared with the CVA group,the index of lung of mice,the OVA-sIgE level in BALF and expression levels of Th2-related factor in the lung tissue of mice in Prednisone Acetate and Glycyrrhetinic Acid groups were significantly decreased(P<0.05 or P<0.01),the infiltration of inflammatory cells in the lung tissue was reduced,while expressions of Th1-related factor in the lung tissue was significantly increased(P<0.05 or P<0.01).Conclusion:Glycyrrhetinic acid has therapeutic effect on CVA mice,the underlying mechanism of Glycyrrhetinic acid alleviating lung impairment and airway inflammation may be associated with mediating the Th1/Th2 imbalance in the lung tissue.展开更多
Objective: To study the influence of glycyrrhetinic acid(GA) on bronchial asthma(BA)smooth muscle proliferation and apoptosis as well as inflammatory factor expression and its molecular mechanism.Methods: Male SD guin...Objective: To study the influence of glycyrrhetinic acid(GA) on bronchial asthma(BA)smooth muscle proliferation and apoptosis as well as inflammatory factor expression and its molecular mechanism.Methods: Male SD guinea pigs were selected and made into asthma models, bronchial asthma smooth muscle cells were cultured and divided into BA group, GA group and GA + LM group that were treated with serum-free RPMI1640 culture medium, serumfree RPMI1640 culture medium containing 50 ng/mL glycyrrhetinic acid, serum-free RPMI1640 culture medium containing 50 ng/mL glycyrrhetinic acid and 100 ng/mL LM22B-10 respectively; normal guinea pigs were collected and bronchial smooth muscle cells were cultured as control group. The cell proliferation activity as well as the expression of proliferation and apoptosis genes, inflammatory factors and p-ERK1/2 was determined.Results: Proliferation activity value and m RNA expression of Bcl-2, TNF-α, IL-4, IL-6,YKL-40, protein expression of p-ERK1/2 of airway smooth muscle cell in BA group were significantly higher than those of control group while m RNA expression levels of Bax,caspase-9 as well as caspase-3 were significantly lower than that of control group(P < 0.05); proliferation activity value and m RNA expression of Bcl-2, TNF-α, IL-4, IL-6, YKL-40, protein expression of p-ERK1/2 of airway smooth muscle cell in GA group were significantly lower than those of BA group(P < 0.05) while the m RNA expression levels of Bax, caspase-9 as well as caspase-3 were significantly higher than those of BA group(P < 0.05); proliferation activity value and m RNA expression of Bcl-2, TNF-α, IL-4, IL-6, YKL-40 of airway smooth muscle cell in GA + LM group were significantly higher than those of GA group(P < 0.05) while m RNA expression levels of Bax, caspase-9 as well as caspase-3 were significantly lower that of GA group(P < 0.05).Conclusion: GA can inhibit the proliferation of bronchial smooth muscle cells and reduce the expression of inflammatory factors by inhibiting the phosphorylation of ERK1/2.展开更多
Background: Airway inflammation is the core pathological process of asthma, with the key inflammatory regulators incompletely defined. Recently, fibroblast growth factor 2(FGF2) has been reported to be an inflammatory...Background: Airway inflammation is the core pathological process of asthma, with the key inflammatory regulators incompletely defined. Recently, fibroblast growth factor 2(FGF2) has been reported to be an inflammatory regulator;however, its role in asthma remains elusive. This study aimed to investigate the immunomodulatory role of FGF2 in asthma.Methods: First, FGF2 expression was characterised in clinical asthma samples and the house dust mite(HDM)-induced mouse chronic asthma model. Second, recombinant mouse FGF2(rm-FGF2) protein was intranasally delivered to determine the effect of FGF2 on airway inflammatory cell infiltration. Third, human airway epithelium-derived A549 cells were stimulated with either HDM or recombinant human interleukin-1β(IL-1β) protein combined with or without recombinant human FGF2. IL-1β-induced IL-6 or IL-8 release levels were determined using enzyme-linked immunosorbent assay, and the involved signalling transduction was explored via Western blotting.Results: Compared with the control groups, the FGF2 protein levels were significantly upregulated in the bronchial epithelium and alveolar areas of clinical asthma samples [(6.70±1.79) vs.(16.32±2.40), P=0.0184;(11.20±2.11) vs.(21.00±3.00), P=0.033, respectively] and HDM-induced asthmatic mouse lung lysates [(1.00±0.15) vs.(5.14±0.42),P<0.001]. Moreover, FGF2 protein abundance was positively correlated with serum total and anti-HDM IgE levels in the HDM-induced chronic asthma model(R^(2)=0.857 and 0.783, P=0.0008 and 0.0043, respectively). Elevated FGF2protein was mainly expressed in asthmatic bronchial epithelium and alveolar areas and partly co-localised with infiltrated inflammatory cell populations in HDM-induced asthmatic mice. More importantly, intranasal instillation of rm-FGF2 aggravated airway inflammatory cell infiltration [(2.45±0.09) vs.(2.88±0.14), P=0.0288] and recruited more subepithelial neutrophils after HDM challenge [(110.20±29.43) cells/mm^(2) vs.(238.10±42.77) cells/mm^(2), P=0.0392]without affecting serum IgE levels and Th2 cytokine transcription. In A549 cells, FGF2 was upregulated through HDM stimulation and promoted IL-1β-induced IL-6 or IL-8 release levels [up to(1.41±0.12)-or(1.44±0.14)-fold change vs.IL-1β alone groups, P=0.001 or 0.0344, respectively]. The pro-inflammatory effect of FGF2 is likely mediated through the fibroblast growth factor receptor(FGFR)/mitogen-activated protein kinase(MAPK)/nuclear factor kappa B(NF-κB)pathway.Conclusions: Our findings suggest that FGF2 is a potential inflammatory modulator in asthma, which can be induced by HDM and acts through the FGFR/MAPK/NF-κB pathway in the airway epithelial cells.展开更多
This study examined the expression of the anterior gradient-2 （AGR2） protein and Muc5ac protein in the lung tissues of asthmatic mice and the effect of dexamethasone, with an at- tempt to explore the role of AGR2 in...This study examined the expression of the anterior gradient-2 （AGR2） protein and Muc5ac protein in the lung tissues of asthmatic mice and the effect of dexamethasone, with an at- tempt to explore the role of AGR2 in the over-secretion of mucus in the airway. Eighteen BALB/c mice were divided into asthma group, control group and dexamethasone group. In dexamethasone group, dexamethasone was intraperitoneally administered. Expression of AGR2 protein and Muc5ac protein in the murine lung tissues was immunohistochemically detected. IL-13 level was determined in the bronchoalveolar lavage fluid （BALF） by ELISA. The results exhibited that the expression of AGR2 protein in asthma group （0.522±0.041） was significantly higher than that in normal controls （0.361±0.047） （P〈0.01） and bore a positive linear relationship to the expression of Muc5ac protein （r=0.873, P〈0.05） and IL-13 level （r=0.828, P〈0.05）. Expression of AGR2 protein in the dexa- methasone group （0.456±0.049） was significantly lower than that in the asthma group. It was concluded that： （1） the expression of AGR2 protein was significantly higher in asthmatic mice as com- pared with their normal counterparts; （2） the expression was obviously related to the expression of Muc5ac protein and IL-13; （3） dexamethasone could down-regulate the expression of AGR2 protein. Our findings suggested that AGR2 might be involved in the over-secretion of mucus in the airway in asthma.展开更多
Objective: To explore the effect of PPAR γ agonist(rosiglitazone) on the secretion of Th2 cytokines and the proportion of immune cell subsets in asthma mice,Methods: Ovalbumin(OVA)-sensitized mice were used to build ...Objective: To explore the effect of PPAR γ agonist(rosiglitazone) on the secretion of Th2 cytokines and the proportion of immune cell subsets in asthma mice,Methods: Ovalbumin(OVA)-sensitized mice were used to build asthma models,Those mice were divided into the normal control group,model group and rosiglitazone group,Differences of the changes in lung histopathology of mice in the three group were observed through hematoxylin and eosin(HE) strain,and the numbers of the total cells,eosinophils and neutrophils in BALF of mice in the three groups were compared,ELISA and real-time PCR were employed to detect the protein levels of interleukin(IL)-5,IL-13,IL-4 and IL-10 and m RNA level,respectively,Flow cytometry number was implied to analyze the proportion of immune cell subsets in peripheral blood of mice,Results: Compared with the mice in the control group,and mice of the model group,the infiltration of inflammatory cells in BALF increased,bronchial smooth muscle became thickened,a large amount of collagen deposited,the secretion of Th2 cytokine increased significantly,the ratio of regulatory T cells(Treg) decreased,the ratio of T17 cells rose distinctly; while in mice of the rosiglitazone group,the changes of their lung histopathology were improved obviously,the number of infiltration of inflammatory cells declined,the thickened smooth muscle relieved,the deposition of collagen decreased,the secretion of Th2 cytokine was inhibited,the ratio of Treg went up,and the increased of the ratio of T17 cells was inhibited but still not return to normal level,Conclusions: Rosiglitazone can regulate the proportion of Treg and Th17 cells and inhibit the secretion of Th2 cytokines,which inhibit the airway inflammatory response for asthma mice effectively.展开更多
BACKGROUND： Several studies have demonstrated that SH2-B13 is over-expressed in the lung, C7T5 spinal ganglia, and the corresponding spinal dorsal horn of asthmatic mice. SH2-Bβ expression has been shown to positive...BACKGROUND： Several studies have demonstrated that SH2-B13 is over-expressed in the lung, C7T5 spinal ganglia, and the corresponding spinal dorsal horn of asthmatic mice. SH2-Bβ expression has been shown to positively correlate with nerve growth factor （NGF） expression levels. This indicates that SH2-Bβ, in the presence of NGF, may participate in asthmatic attacks. OBJECTIVE： To observe the effects of anti-NGF on SH2-Bβ expression in primary afferent neurons （C7-T5 spinal ganglia and corresponding spinal dorsal horn） and in the lung to further investigate the regulatory effects of NGF on SH2-B/3 expression. DESIGN, TIME AND SETTING： A completely randomized block design experiment was performed at the Department of Neurobiology, China Medical University between March 2004 and July 2005. MATERIALS： Thirty-six male, BALB/c mice were included in this study. Ovalbumin solution was purchased from Sigma, USA. SH2-Bβ polyclonal antibody was provided by Santa Cruz, USA. Anti-NGF reagent was obtained from Wuhan Boster Bioengineering Co., Ltd., China. METHODS： Thirty-six mice were randomly and evenly divided into three groups： control, model, and anti-NGF. In the model group, asthma was induced by intraperitoneal injection and aerosol inhalation of ovalbumin solution. Mice in the anti-NGF group received anti-NGF through the nasal cavity 3 hours prior to aerosol inhalation. In the control group, mice were subjected to experimental procedures similar to the model group, except that ovalbumin solution was replaced by phosphate buffered saline （PBS）. MAIN OUTCOME MEASURES： SH2-Bβ expression in primary afferent neurons （C7T5 spinal ganglia and the corresponding spinal dorsal horn） and the lung was detected by immunohistochemistry and Western blot. Immunostaining intensity level of SH2-Bβ was analyzed using the MetaMorph image analysis system. RESULTS： lmmunohistochemistry results revealed that the mean intensity of SH2-Bβ expression in the C7 T5 spinal ganglia, the corresponding spinal dorsal horn, and the lungs was significantly greater in the model group than in the control group （P 〈 0.01）. However, expression was significantly less in the anti-NGF group, compared with the model group （P 〈 0.01）. Western Blot results demonstrated that SH2-Bβ expression was significantly greater in the model group C7-T5 spinal ganglia and lung, compared with the control group （P 〈 0.01）. However, expression was significantly less in the anti-NGF group, compared with the model group （P 〈 0.01）. CONCLUSION： The present study showed that, in the primary afferent neurons and the lung, SH2-Bβ participated in asthmatic attack. Anti-NGF down-regulated SH2-Bβ expression in C7-T5 spinal ganglia and the corresponding spinal dorsal horn, as well as the lung, of asthmatic mice. These results indicate that SH2-Bβ could be an important signaling molecule in mediating effects of NGF during asthmatic attack.展开更多
Four regulatory single nucleotide polymorphisms (rSNPs) (rs2238631, rs2238632, rs2238633 and rs2238634) in intron one, two rSNPs (rs1131882 and rs4523) in exon 3 and one rSNP (rs5756) in the 3’UTR of the thromboxane ...Four regulatory single nucleotide polymorphisms (rSNPs) (rs2238631, rs2238632, rs2238633 and rs2238634) in intron one, two rSNPs (rs1131882 and rs4523) in exon 3 and one rSNP (rs5756) in the 3’UTR of the thromboxane A2 receptor (TBXA2R) gene have been associated with childhood-onset asthma in Asians. These rSNP alleles alter the DNA landscape for potential transcriptional factors (TFs) to attach resulting in changes in transcriptional factor binding sites (TFBS). These TFBS changes are examined with respect to asthma which has been found to be significantly associated with the rSNPs.展开更多
基金The Sixth Batch of Special Support Plans in Anhui Province(No.dlPtzjh20200050)Key Natural Science Research Project of Higher Education Institutions in Anhui Province(No.KJ2020A0426)。
文摘Objective: To investigate the effects of Yanghe Pingchuan Granules on airway remodeling in asthmatic rats, and to explore the mechanism of Interleukin-6/Janus kinase 2/ Signal transducing activator of transcription 3(IL-6/JAK2/STAT3) signal axis. Methods: We separated 42 healthy male SD rats into two groups, a control group (7) and a model group (35).The model group was sensitized with a combination of ovalbumin (OVA) and aluminum hydroxide for 2 weeks, while the control group was given an equal amount of physiological saline.After 2 weeks, the modeling group was randomly divided into Model group, Yanghe Pingchuan Granules high, medium and low dose groups and Dexamethasone group, each group consisted of 7 animals. After 4 weeks, OVA atomization and gavage were used for stimulation and treatment. Yanghe Pingchuan Granules high, middle and low groups were given 15.48, 7.74, 3.87 g∙kg-1 Yanghe Pingchuan Granules daily, dexamethasone group was given 0.0625 mg∙kg-1 dexamethasone daily, and the other groups were given the same amount of normal saline. HE, PAS and Masson staining were used to observe the lung histopathological changes in rats. The levels of interleukin-6, IL-23 and IL-17A were detected by ELISA. The expression levels of JAK-2, P-JAK2, STAT3 and P-STAT3 in lung tissues were detected by Western blot. Real-time quantitative polymerase chain reaction (qRT-PCR) was used to detect the mRNA expression levels of IL-6, JAK2 and STAT3 in rat lung tissue. Results: The lung tissue structure of the model group was severely damaged compared to the control group, accompanied by a great many of inflammatory cell infiltration, goblet cell hyperplasia, subepithelial collagen fiber deposition and airway epithelial thickening were more obvious. The expressions of IL-6, IL- 23 and IL-17A in serum were significantly increased (P<0.01), the protein expression levels of JAK-2, P-JAK2, STAT3 and P-STAT3 and the mRNA expression levels of IL-6, JAK2 and STAT3 in lung tissue were significantly increased (P<0.01);Compared with the model group, inflammatory cell infiltration, goblet cell proliferation, subepithelial collagen fiber deposition and airway epithelial thickening were significantly reduced in each administration group, and the expressions of IL-6, IL-23 and IL-17A in serum were significantly decreased (P< 0.01). The protein expression levels of JAK-2, P-JAK2, STAT3 and P-STAT3 and mRNA expression levels of IL-6, JAK2 and STAT3 in lung tissue were significantly decreased (P<0.01). Conclusion: Yanghe Pingchuan Granules can significantly alleviate airway remodeling in asthmatic rats, and its mechanism may be through inhibiting the IL-6/JAK2/STAT3 signal axis.
基金General Projects of National Natural Science Fund(81873338)Key Scientific and Technological Projects in Henan Province(202102310491)Training Subjects of Leading Talents in the Department of Traditional Chinese Medicine in Henan Province(Yuweizhongyihan[2021]No.8)。
文摘Objective:To establish and verify aβ_(2)-AR desensitization asthma mice model.Methods:A total of 30 SPF male BALB/c mice were randomly divided into blank group,the common asthma group,andβ_(2)-AR desensitization asthma model group.Asthma model was established,and on this basis,the method of atom-izing inhalation and intraperitoneal injections of salbutamol was used to prepareβ_(2)-AR desensitization asthma model.After the last stimulation on the 21st day of modeling,the airway resistance of mice was measured.ELISA was used to detect the content of serum IgE;HE staining was used to observe the lung organization degree of infla-mmatory cell infiltration;Western blot method was used to detect theβ_(2)-AR content in lung tissue,RT-PCR was used to detect theβ_(2)-ARmRNA expressionin lung tissue.Results:Compared with the blank group,as acetyl choline(Mch)levels increased,groups of OVA induced airway resistance increases;but theβ_(2)-AR desensitization asthma model group increased airway resistance was more significant(P<0.05);compared with the blank group,IgE levels of common asthma group andβ_(2)-AR desensitization asthma model group elevated(P<0.01).The pathological histology observation found theβ_(2)-AR desensitization asthma airway inflammation infiltration in mice,the excessive mucus secretion and collagen deposition,and the pathological performance obviously increase compared with the common asthma group;β_(2)-AR content in the lung tissue ofβ_(2)-AR desensitization asthma model in mice,β_(2)-AR mRNA expression level in the blank group and common asthma model group were significantly decreased(P<0.05).Conclusion:Theβ_(2)-AR desensitization asthma mouse model was successfully established,and the buildingcycle was short.
基金National Natural Science Foundation of China (No.81960887)Innovation Project of Guangxi Graduate Education (No.YCXJ2021119)。
文摘Objective:To evaluate the therapeutic effect of Glycyrrhetinic Acid on cough variant asthma(CVA)mice and to investigate the possible mechanism in reducing lung inflammation.Methods:48 young female Balb/c mice were divided into Control,CVA,Prednisone Acetate,Glycyrrhetinic Acid high-dose,Glycyrrhetinic Acid middle-dose and Glycyrrhetinic Acid lowdose groups randomly,with 8 mice in each group.The CVA mice model was established by ovalbumin(OVA)sensitization and OVA challenge,the animal asthma behavior was observed after drug administration,and the index of the lung of mice were recorded.The level of OVAsIgE in the bronchoalveolar lavage fluid(BALF)was tested by ELISA.The pathological changes of the lung tissue were observed by Hematoxylin and Eosin(H&E)staining.The protein expressions of T-bet,IFN-γ,Gata3,IL-4 and IL-13 in the lung tissue were determined by Western blot.Results:Compared with the CVA group,the index of lung of mice,the OVA-sIgE level in BALF and expression levels of Th2-related factor in the lung tissue of mice in Prednisone Acetate and Glycyrrhetinic Acid groups were significantly decreased(P<0.05 or P<0.01),the infiltration of inflammatory cells in the lung tissue was reduced,while expressions of Th1-related factor in the lung tissue was significantly increased(P<0.05 or P<0.01).Conclusion:Glycyrrhetinic acid has therapeutic effect on CVA mice,the underlying mechanism of Glycyrrhetinic acid alleviating lung impairment and airway inflammation may be associated with mediating the Th1/Th2 imbalance in the lung tissue.
基金supported by Guangdong Medical Science and Technology Research Fund Project(No:A2017331)
文摘Objective: To study the influence of glycyrrhetinic acid(GA) on bronchial asthma(BA)smooth muscle proliferation and apoptosis as well as inflammatory factor expression and its molecular mechanism.Methods: Male SD guinea pigs were selected and made into asthma models, bronchial asthma smooth muscle cells were cultured and divided into BA group, GA group and GA + LM group that were treated with serum-free RPMI1640 culture medium, serumfree RPMI1640 culture medium containing 50 ng/mL glycyrrhetinic acid, serum-free RPMI1640 culture medium containing 50 ng/mL glycyrrhetinic acid and 100 ng/mL LM22B-10 respectively; normal guinea pigs were collected and bronchial smooth muscle cells were cultured as control group. The cell proliferation activity as well as the expression of proliferation and apoptosis genes, inflammatory factors and p-ERK1/2 was determined.Results: Proliferation activity value and m RNA expression of Bcl-2, TNF-α, IL-4, IL-6,YKL-40, protein expression of p-ERK1/2 of airway smooth muscle cell in BA group were significantly higher than those of control group while m RNA expression levels of Bax,caspase-9 as well as caspase-3 were significantly lower than that of control group(P < 0.05); proliferation activity value and m RNA expression of Bcl-2, TNF-α, IL-4, IL-6, YKL-40, protein expression of p-ERK1/2 of airway smooth muscle cell in GA group were significantly lower than those of BA group(P < 0.05) while the m RNA expression levels of Bax, caspase-9 as well as caspase-3 were significantly higher than those of BA group(P < 0.05); proliferation activity value and m RNA expression of Bcl-2, TNF-α, IL-4, IL-6, YKL-40 of airway smooth muscle cell in GA + LM group were significantly higher than those of GA group(P < 0.05) while m RNA expression levels of Bax, caspase-9 as well as caspase-3 were significantly lower that of GA group(P < 0.05).Conclusion: GA can inhibit the proliferation of bronchial smooth muscle cells and reduce the expression of inflammatory factors by inhibiting the phosphorylation of ERK1/2.
基金supported by grants awarded to YY by the National Natural Science Foundation of China (81870019, 82170029)the Guangdong Provincial Natural Science Foundation (2018A030313554)+3 种基金the Innovation Research Team for Basic and Clinical Studies on Chronic Liver Diseases of 2018 High-Level Health Teams of ZhuhaiYKQ by the National Natural Science Foundation of China (82002612)the Chinese Postdoctoral Science Foundation (2019M660211)ZGC by the Science and Technology Program of Guangzhou,China (201704020179)。
文摘Background: Airway inflammation is the core pathological process of asthma, with the key inflammatory regulators incompletely defined. Recently, fibroblast growth factor 2(FGF2) has been reported to be an inflammatory regulator;however, its role in asthma remains elusive. This study aimed to investigate the immunomodulatory role of FGF2 in asthma.Methods: First, FGF2 expression was characterised in clinical asthma samples and the house dust mite(HDM)-induced mouse chronic asthma model. Second, recombinant mouse FGF2(rm-FGF2) protein was intranasally delivered to determine the effect of FGF2 on airway inflammatory cell infiltration. Third, human airway epithelium-derived A549 cells were stimulated with either HDM or recombinant human interleukin-1β(IL-1β) protein combined with or without recombinant human FGF2. IL-1β-induced IL-6 or IL-8 release levels were determined using enzyme-linked immunosorbent assay, and the involved signalling transduction was explored via Western blotting.Results: Compared with the control groups, the FGF2 protein levels were significantly upregulated in the bronchial epithelium and alveolar areas of clinical asthma samples [(6.70±1.79) vs.(16.32±2.40), P=0.0184;(11.20±2.11) vs.(21.00±3.00), P=0.033, respectively] and HDM-induced asthmatic mouse lung lysates [(1.00±0.15) vs.(5.14±0.42),P<0.001]. Moreover, FGF2 protein abundance was positively correlated with serum total and anti-HDM IgE levels in the HDM-induced chronic asthma model(R^(2)=0.857 and 0.783, P=0.0008 and 0.0043, respectively). Elevated FGF2protein was mainly expressed in asthmatic bronchial epithelium and alveolar areas and partly co-localised with infiltrated inflammatory cell populations in HDM-induced asthmatic mice. More importantly, intranasal instillation of rm-FGF2 aggravated airway inflammatory cell infiltration [(2.45±0.09) vs.(2.88±0.14), P=0.0288] and recruited more subepithelial neutrophils after HDM challenge [(110.20±29.43) cells/mm^(2) vs.(238.10±42.77) cells/mm^(2), P=0.0392]without affecting serum IgE levels and Th2 cytokine transcription. In A549 cells, FGF2 was upregulated through HDM stimulation and promoted IL-1β-induced IL-6 or IL-8 release levels [up to(1.41±0.12)-or(1.44±0.14)-fold change vs.IL-1β alone groups, P=0.001 or 0.0344, respectively]. The pro-inflammatory effect of FGF2 is likely mediated through the fibroblast growth factor receptor(FGFR)/mitogen-activated protein kinase(MAPK)/nuclear factor kappa B(NF-κB)pathway.Conclusions: Our findings suggest that FGF2 is a potential inflammatory modulator in asthma, which can be induced by HDM and acts through the FGFR/MAPK/NF-κB pathway in the airway epithelial cells.
基金supported by the National Natural Science Foundation of China (No. 30900648)
文摘This study examined the expression of the anterior gradient-2 （AGR2） protein and Muc5ac protein in the lung tissues of asthmatic mice and the effect of dexamethasone, with an at- tempt to explore the role of AGR2 in the over-secretion of mucus in the airway. Eighteen BALB/c mice were divided into asthma group, control group and dexamethasone group. In dexamethasone group, dexamethasone was intraperitoneally administered. Expression of AGR2 protein and Muc5ac protein in the murine lung tissues was immunohistochemically detected. IL-13 level was determined in the bronchoalveolar lavage fluid （BALF） by ELISA. The results exhibited that the expression of AGR2 protein in asthma group （0.522±0.041） was significantly higher than that in normal controls （0.361±0.047） （P〈0.01） and bore a positive linear relationship to the expression of Muc5ac protein （r=0.873, P〈0.05） and IL-13 level （r=0.828, P〈0.05）. Expression of AGR2 protein in the dexa- methasone group （0.456±0.049） was significantly lower than that in the asthma group. It was concluded that： （1） the expression of AGR2 protein was significantly higher in asthmatic mice as com- pared with their normal counterparts; （2） the expression was obviously related to the expression of Muc5ac protein and IL-13; （3） dexamethasone could down-regulate the expression of AGR2 protein. Our findings suggested that AGR2 might be involved in the over-secretion of mucus in the airway in asthma.
基金Supported by medical research project of Health and Family Planning Commission of Chongqing(Grant No.20142019)
文摘Objective: To explore the effect of PPAR γ agonist(rosiglitazone) on the secretion of Th2 cytokines and the proportion of immune cell subsets in asthma mice,Methods: Ovalbumin(OVA)-sensitized mice were used to build asthma models,Those mice were divided into the normal control group,model group and rosiglitazone group,Differences of the changes in lung histopathology of mice in the three group were observed through hematoxylin and eosin(HE) strain,and the numbers of the total cells,eosinophils and neutrophils in BALF of mice in the three groups were compared,ELISA and real-time PCR were employed to detect the protein levels of interleukin(IL)-5,IL-13,IL-4 and IL-10 and m RNA level,respectively,Flow cytometry number was implied to analyze the proportion of immune cell subsets in peripheral blood of mice,Results: Compared with the mice in the control group,and mice of the model group,the infiltration of inflammatory cells in BALF increased,bronchial smooth muscle became thickened,a large amount of collagen deposited,the secretion of Th2 cytokine increased significantly,the ratio of regulatory T cells(Treg) decreased,the ratio of T17 cells rose distinctly; while in mice of the rosiglitazone group,the changes of their lung histopathology were improved obviously,the number of infiltration of inflammatory cells declined,the thickened smooth muscle relieved,the deposition of collagen decreased,the secretion of Th2 cytokine was inhibited,the ratio of Treg went up,and the increased of the ratio of T17 cells was inhibited but still not return to normal level,Conclusions: Rosiglitazone can regulate the proportion of Treg and Th17 cells and inhibit the secretion of Th2 cytokines,which inhibit the airway inflammatory response for asthma mice effectively.
文摘BACKGROUND： Several studies have demonstrated that SH2-B13 is over-expressed in the lung, C7T5 spinal ganglia, and the corresponding spinal dorsal horn of asthmatic mice. SH2-Bβ expression has been shown to positively correlate with nerve growth factor （NGF） expression levels. This indicates that SH2-Bβ, in the presence of NGF, may participate in asthmatic attacks. OBJECTIVE： To observe the effects of anti-NGF on SH2-Bβ expression in primary afferent neurons （C7-T5 spinal ganglia and corresponding spinal dorsal horn） and in the lung to further investigate the regulatory effects of NGF on SH2-B/3 expression. DESIGN, TIME AND SETTING： A completely randomized block design experiment was performed at the Department of Neurobiology, China Medical University between March 2004 and July 2005. MATERIALS： Thirty-six male, BALB/c mice were included in this study. Ovalbumin solution was purchased from Sigma, USA. SH2-Bβ polyclonal antibody was provided by Santa Cruz, USA. Anti-NGF reagent was obtained from Wuhan Boster Bioengineering Co., Ltd., China. METHODS： Thirty-six mice were randomly and evenly divided into three groups： control, model, and anti-NGF. In the model group, asthma was induced by intraperitoneal injection and aerosol inhalation of ovalbumin solution. Mice in the anti-NGF group received anti-NGF through the nasal cavity 3 hours prior to aerosol inhalation. In the control group, mice were subjected to experimental procedures similar to the model group, except that ovalbumin solution was replaced by phosphate buffered saline （PBS）. MAIN OUTCOME MEASURES： SH2-Bβ expression in primary afferent neurons （C7T5 spinal ganglia and the corresponding spinal dorsal horn） and the lung was detected by immunohistochemistry and Western blot. Immunostaining intensity level of SH2-Bβ was analyzed using the MetaMorph image analysis system. RESULTS： lmmunohistochemistry results revealed that the mean intensity of SH2-Bβ expression in the C7 T5 spinal ganglia, the corresponding spinal dorsal horn, and the lungs was significantly greater in the model group than in the control group （P 〈 0.01）. However, expression was significantly less in the anti-NGF group, compared with the model group （P 〈 0.01）. Western Blot results demonstrated that SH2-Bβ expression was significantly greater in the model group C7-T5 spinal ganglia and lung, compared with the control group （P 〈 0.01）. However, expression was significantly less in the anti-NGF group, compared with the model group （P 〈 0.01）. CONCLUSION： The present study showed that, in the primary afferent neurons and the lung, SH2-Bβ participated in asthmatic attack. Anti-NGF down-regulated SH2-Bβ expression in C7-T5 spinal ganglia and the corresponding spinal dorsal horn, as well as the lung, of asthmatic mice. These results indicate that SH2-Bβ could be an important signaling molecule in mediating effects of NGF during asthmatic attack.
文摘Four regulatory single nucleotide polymorphisms (rSNPs) (rs2238631, rs2238632, rs2238633 and rs2238634) in intron one, two rSNPs (rs1131882 and rs4523) in exon 3 and one rSNP (rs5756) in the 3’UTR of the thromboxane A2 receptor (TBXA2R) gene have been associated with childhood-onset asthma in Asians. These rSNP alleles alter the DNA landscape for potential transcriptional factors (TFs) to attach resulting in changes in transcriptional factor binding sites (TFBS). These TFBS changes are examined with respect to asthma which has been found to be significantly associated with the rSNPs.
