AIM: To investigate the effect of chymase inhibitor TY-51469 in the therapy of inflammatory bowel disease and the underlying mechanism. METHODS: Seventy-five healthy Sprague-Dawley rats were randomly assigned to one o...AIM: To investigate the effect of chymase inhibitor TY-51469 in the therapy of inflammatory bowel disease and the underlying mechanism. METHODS: Seventy-five healthy Sprague-Dawley rats were randomly assigned to one of the three groups(control group, model group and TY-51469 experiment group) and each group had twenty-five rats. The rats of the model group and experiment group were subjected to treatment with 3.5% dextran sulfate sodium(DSS) 10 mg/kg to induce colitis. The control group and model group were subjected to intraperitoneal injection of saline, while the experiment group was subjected to intraperitoneal injection of 10 mg/kg TY-51469 each day. Five rats of each group were sacrificed on 0, 7, 14, 21 and 28 d, respectively. The degree of inflammation was assessed by histopathological scoring; flow cytometry was performed to detect the proportion of CD4+CD25+ Tregs in peripheral blood; colon tissues of rats were collected to measure m RNA and protein expression by PCR, Western blot and immunohistochemistry; serum levels of interleukin(IL)-10, transforming growth factor(TGF)-β1 and IL-17A were detected by ELISA. RESULTS: The rats in the experiment group and model group had significantly more severe colitis than the ones in the control group(P < 0.05) before treatment on day 0; no significant difference was observed between the experiment group and model group(P > 0.05). After treatment with TY-51469, the rats in the experiment group had significantly less severe colitis compared with the model group on 7, 14, 21 and 28 d(P < 0.05). The proportion of CD4+CD25+ Tregs was lower in the model group and experiment group than in the control group; the experiment group had a significantly higher proportion of CD4+CD25+ Tregs than that in the model group(P < 0.05). The model group and experiment group demonstrated lower expression of Foxp3 than the control group; the experiment group had higher Foxp3 expression than the model group(P < 0.05). Cytokines IL-10, TGF-β1 and IL-17 A were lower in the model group and experiment group than in the control group; the experiment group had higher expression than the model group(P < 0.05). CONCLUSION: After treatment with chymase inhibitor TY-51469, the experiment group demonstrated more significantly reduced intestinal inflammation and higher expression of immune tolerance related cytokines(IL-10, TGF-β1, IL-17A) and Foxp3 which is specifically expressed in Tregs compared with the model group. Therefore, chymase inhibitor TY-51469 might ameliorate the progression of DSS-induced colitis possibly by increasing the expression of Tregs and cytokines.展开更多
Background Little is known about the role of dual angiotensin Ⅱ forming pathways during heart failure. In the present study, the changes of chymase and angiotensin converting enzyme （ACE） expressions in the failing...Background Little is known about the role of dual angiotensin Ⅱ forming pathways during heart failure. In the present study, the changes of chymase and angiotensin converting enzyme （ACE） expressions in the failing hearts of hamsters were analysed. Methods Heart failure was induced by ligation of left anterior descending branch of the coronary artery. Chymase, ACE and angiotensin Ⅱ type 1 receptor （AT1R） mRNA levels were analysed by reverse transcription polymerase chain reaction （RT-PCR）. The activities of chymase and ACE were determined by radioimmunoassay （RIA）. Myocardial collagen fibre analysis was performed under optical microscope. Results Left ventricular systolic pressure （LVSP） and maximum left ventricular developed pressure increase rate （ dp/dtmax, mmHg/s） gradually moved lower at 2, 3,4 and 8 weeks after operation. On the other hand, left ventricular end-diastolic pressure （LVEDP） increased gradually after operation. Compared with the control group （3.55±0. 06, 4.79±0.70）, the heart weight/body weight ratio in operation group had increased significantly at 4 weeks and 8 weeks （4. 28±0. 43, 6. 17±0.73） （P 〈0. 01 ）. Collagen staining showed that the quantity of myocardial collagen fibre increased significantly in the operation group. RT-PCR showed that the chymase mRNA level in the operation group was consistently greater than that in the control group. ATIR mRNA level was also increased significantly at 3 weeks and 4 weeks, both being 1.3 times that of the control group （ P〈0.01 ）, whereas ACE mRNA level was not changed. Higher activity of chymase was detected in operation group, being 4, 8, 13 and 19 times that of the control group at 2, 3, 4 and 8 weeks （P〈0.01 ）, respectively. ACE activity was also significantly higher at the same time, being 7, 10, 10 and 3.5 times that of the control（P〈0.01）. Angiotensin Ⅱ （Ang Ⅱ） level in operation group increased significantly, being 2.5, 2.7, 3.5 and 2 times that of the control group at 2, 3, 4 and 8 weeks, respectively （P 〈 0. 01 ）. Conclusions A dual Ang Ⅱ forming pathway from both ACE and chymase in the hamster hearts plays an important role during the development of heart failure. At the decompensatory stage, the reduction of AngⅡ level may be associated with the decrease of ACE activity.展开更多
The myosin light chain 2 promoter-human heart chymase (MLC2-hChymase) transgenic mice founded by our laboratory were used as the model to study the function of chymase in the heart angiotension Ⅱ (Ang Ⅱ) formation a...The myosin light chain 2 promoter-human heart chymase (MLC2-hChymase) transgenic mice founded by our laboratory were used as the model to study the function of chymase in the heart angiotension Ⅱ (Ang Ⅱ) formation and heart remodeling. Tissue-specific expression of human heart chymase gene and transcriptional expression of type Ⅰ and type Ⅲ collagens genes were analyzed by RT-PCR. Activities of chymase, ACE and the levels of Ang Ⅱ in heart and plasma were determined with radioimmunoas-say (RIA) kit. Activity of heart matrix metalloproteinase-9 (MMP-9) was detected using gelatin zymography. The cardiac hypertrophic phenotypes were also observed with the physiological and morphological methods. The results in the MLC2-hChymase transgenic mice indicated: ( i ) human heart chymase gene was expressed specially in the heart; (u) heart chymase activity increased markedly in the transgenic mice vs non-transgenic mice (control) (0.27±0.07 U/mg vs. 0.15±0.02 U/mg, P【0.05) with no significant difference in展开更多
Mast cells (MCs) play a pivotal role in the hypersensitivity reaction by regulating the innate and adaptive immune responses. Humans have two types of MCs. The first type, termed MCTC, is found in the skin and other c...Mast cells (MCs) play a pivotal role in the hypersensitivity reaction by regulating the innate and adaptive immune responses. Humans have two types of MCs. The first type, termed MCTC, is found in the skin and other connective tissues and expresses both tryptase and chymase, while the second, termed MCT, which only expresses tryptase, is found primarily in the mucosa. MCs induced from human adult-type CD34+ cells are reported to be of the MCT type, but the development of MCs during embryonic/fetal stages is largely unknown. Using an efficient coculture system, we identified that a CD34+c-kit+ cell population, which appeared prior to the emergence of CD34+CD45+ hematopoietic stem and progenitor cells (HSPCs), stimulated robust production of pure Tryptase+Chymase+ MCs (MCTCs). Single-cell analysis revealed dual development directions of CD34+c-kit+ progenitors, with one lineage developing into erythro-myeloid progenitors (EMP) and the other lineage developing into HSPC. Interestingly, MCTCs derived from early CD34+c-kit+ cells exhibited strong histamine release and immune response functions. Particularly, robust release of IL-17 suggested that these early developing tissue-type MCTCs could play a central role in tumor immunity. These findings could help elucidate the mechanisms controlling early development of MCTCs and have significant therapeutic implications.展开更多
Objective:The gene for mast cell chymase (CMA1) is an ideal candidate for investigating the genetic predisposition to coronary heart disease (CHD),as activated mast cells have been found to be present in a greater pro...Objective:The gene for mast cell chymase (CMA1) is an ideal candidate for investigating the genetic predisposition to coronary heart disease (CHD),as activated mast cells have been found to be present in a greater proportion in the shoulder region of atheroma than in normal coronary intimae.Previous studies have indicated that CMA1 promoter polymorphism rs1800875 may be involved in regulating immunoglobulin E (IgE) levels in patients with eczema,and it is associated with the progression of immunoglobulin A nephropathy.Methods:The association between single nucleotide polymorphism (SNP) rs1800875,serum chymase,and serum IgE levels was examined in 175 CHD subjects and 95 non-CHD subjects.Results:Statistical analysis indicated no significant difference in allele frequency between CHD and non-CHD.However,a significant association was found between CMA1 genotypes and total IgE levels in CHD subjects.Meanwhile,crossover analysis revealed that,in GG homozygotes,CHD risk was nearly six times higher in those with IgE (U/ml) level <2.58 (natural logarithm conversion),while no association was found with chymase level.Conclusions:Polymorphism rs1800875 of CMA1 may be associated with serum IgE level in CHD subjects,but not with chymase level in both groups.In GG homozygotes,high IgE level is a protective factor against coronary disease.展开更多
基金Supported by Science and Technology Project of Liaoning ProvinceNo.2013225303+1 种基金Science and Technology Project of Shenyang CityNo.F13-316-1-40
文摘AIM: To investigate the effect of chymase inhibitor TY-51469 in the therapy of inflammatory bowel disease and the underlying mechanism. METHODS: Seventy-five healthy Sprague-Dawley rats were randomly assigned to one of the three groups(control group, model group and TY-51469 experiment group) and each group had twenty-five rats. The rats of the model group and experiment group were subjected to treatment with 3.5% dextran sulfate sodium(DSS) 10 mg/kg to induce colitis. The control group and model group were subjected to intraperitoneal injection of saline, while the experiment group was subjected to intraperitoneal injection of 10 mg/kg TY-51469 each day. Five rats of each group were sacrificed on 0, 7, 14, 21 and 28 d, respectively. The degree of inflammation was assessed by histopathological scoring; flow cytometry was performed to detect the proportion of CD4+CD25+ Tregs in peripheral blood; colon tissues of rats were collected to measure m RNA and protein expression by PCR, Western blot and immunohistochemistry; serum levels of interleukin(IL)-10, transforming growth factor(TGF)-β1 and IL-17A were detected by ELISA. RESULTS: The rats in the experiment group and model group had significantly more severe colitis than the ones in the control group(P < 0.05) before treatment on day 0; no significant difference was observed between the experiment group and model group(P > 0.05). After treatment with TY-51469, the rats in the experiment group had significantly less severe colitis compared with the model group on 7, 14, 21 and 28 d(P < 0.05). The proportion of CD4+CD25+ Tregs was lower in the model group and experiment group than in the control group; the experiment group had a significantly higher proportion of CD4+CD25+ Tregs than that in the model group(P < 0.05). The model group and experiment group demonstrated lower expression of Foxp3 than the control group; the experiment group had higher Foxp3 expression than the model group(P < 0.05). Cytokines IL-10, TGF-β1 and IL-17 A were lower in the model group and experiment group than in the control group; the experiment group had higher expression than the model group(P < 0.05). CONCLUSION: After treatment with chymase inhibitor TY-51469, the experiment group demonstrated more significantly reduced intestinal inflammation and higher expression of immune tolerance related cytokines(IL-10, TGF-β1, IL-17A) and Foxp3 which is specifically expressed in Tregs compared with the model group. Therefore, chymase inhibitor TY-51469 might ameliorate the progression of DSS-induced colitis possibly by increasing the expression of Tregs and cytokines.
基金This investigation was supported by Major National Basic ResearchProgram, People’s Republic of China (973 Program) (No.G2000056904)
文摘Background Little is known about the role of dual angiotensin Ⅱ forming pathways during heart failure. In the present study, the changes of chymase and angiotensin converting enzyme （ACE） expressions in the failing hearts of hamsters were analysed. Methods Heart failure was induced by ligation of left anterior descending branch of the coronary artery. Chymase, ACE and angiotensin Ⅱ type 1 receptor （AT1R） mRNA levels were analysed by reverse transcription polymerase chain reaction （RT-PCR）. The activities of chymase and ACE were determined by radioimmunoassay （RIA）. Myocardial collagen fibre analysis was performed under optical microscope. Results Left ventricular systolic pressure （LVSP） and maximum left ventricular developed pressure increase rate （ dp/dtmax, mmHg/s） gradually moved lower at 2, 3,4 and 8 weeks after operation. On the other hand, left ventricular end-diastolic pressure （LVEDP） increased gradually after operation. Compared with the control group （3.55±0. 06, 4.79±0.70）, the heart weight/body weight ratio in operation group had increased significantly at 4 weeks and 8 weeks （4. 28±0. 43, 6. 17±0.73） （P 〈0. 01 ）. Collagen staining showed that the quantity of myocardial collagen fibre increased significantly in the operation group. RT-PCR showed that the chymase mRNA level in the operation group was consistently greater than that in the control group. ATIR mRNA level was also increased significantly at 3 weeks and 4 weeks, both being 1.3 times that of the control group （ P〈0.01 ）, whereas ACE mRNA level was not changed. Higher activity of chymase was detected in operation group, being 4, 8, 13 and 19 times that of the control group at 2, 3, 4 and 8 weeks （P〈0.01 ）, respectively. ACE activity was also significantly higher at the same time, being 7, 10, 10 and 3.5 times that of the control（P〈0.01）. Angiotensin Ⅱ （Ang Ⅱ） level in operation group increased significantly, being 2.5, 2.7, 3.5 and 2 times that of the control group at 2, 3, 4 and 8 weeks, respectively （P 〈 0. 01 ）. Conclusions A dual Ang Ⅱ forming pathway from both ACE and chymase in the hamster hearts plays an important role during the development of heart failure. At the decompensatory stage, the reduction of AngⅡ level may be associated with the decrease of ACE activity.
基金the National NaturalScience Foundation of China (Grant No. 39570297), the Foundation of Chinese Academy of Medical Sciences (Grant No. 1995-1997) and in part by the National Major Basic Research Program of China (Grant No. G2000056904).
文摘The myosin light chain 2 promoter-human heart chymase (MLC2-hChymase) transgenic mice founded by our laboratory were used as the model to study the function of chymase in the heart angiotension Ⅱ (Ang Ⅱ) formation and heart remodeling. Tissue-specific expression of human heart chymase gene and transcriptional expression of type Ⅰ and type Ⅲ collagens genes were analyzed by RT-PCR. Activities of chymase, ACE and the levels of Ang Ⅱ in heart and plasma were determined with radioimmunoas-say (RIA) kit. Activity of heart matrix metalloproteinase-9 (MMP-9) was detected using gelatin zymography. The cardiac hypertrophic phenotypes were also observed with the physiological and morphological methods. The results in the MLC2-hChymase transgenic mice indicated: ( i ) human heart chymase gene was expressed specially in the heart; (u) heart chymase activity increased markedly in the transgenic mice vs non-transgenic mice (control) (0.27±0.07 U/mg vs. 0.15±0.02 U/mg, P【0.05) with no significant difference in
基金This work was supported by the National Basic Research Program(973 Program2015CB96A902)+4 种基金the National Natural Science Foundation of China(H81170466 and H81370597)and the CAMS Initiatives for Innovative Medicine(2016-I2M-1-018)awarded to F.M.the CAMS Initiatives for Innovative Medicine(2017-12M-2005)the Union Youth Fund of Chinese Academy of Medical Sciences(81572089)to G.B.and the National Nature Science Foundation of China Youth Fund(81700107)to B.M.
文摘Mast cells (MCs) play a pivotal role in the hypersensitivity reaction by regulating the innate and adaptive immune responses. Humans have two types of MCs. The first type, termed MCTC, is found in the skin and other connective tissues and expresses both tryptase and chymase, while the second, termed MCT, which only expresses tryptase, is found primarily in the mucosa. MCs induced from human adult-type CD34+ cells are reported to be of the MCT type, but the development of MCs during embryonic/fetal stages is largely unknown. Using an efficient coculture system, we identified that a CD34+c-kit+ cell population, which appeared prior to the emergence of CD34+CD45+ hematopoietic stem and progenitor cells (HSPCs), stimulated robust production of pure Tryptase+Chymase+ MCs (MCTCs). Single-cell analysis revealed dual development directions of CD34+c-kit+ progenitors, with one lineage developing into erythro-myeloid progenitors (EMP) and the other lineage developing into HSPC. Interestingly, MCTCs derived from early CD34+c-kit+ cells exhibited strong histamine release and immune response functions. Particularly, robust release of IL-17 suggested that these early developing tissue-type MCTCs could play a central role in tumor immunity. These findings could help elucidate the mechanisms controlling early development of MCTCs and have significant therapeutic implications.
基金Project supported by the National Natural Science Foundation of China (No. 30670867) to Mei-xiang XIANGthe Major Program of Science and Technology Department of Zhejiang Province,China(No. 2007C13058) to Mei-xiang XIANG
文摘Objective:The gene for mast cell chymase (CMA1) is an ideal candidate for investigating the genetic predisposition to coronary heart disease (CHD),as activated mast cells have been found to be present in a greater proportion in the shoulder region of atheroma than in normal coronary intimae.Previous studies have indicated that CMA1 promoter polymorphism rs1800875 may be involved in regulating immunoglobulin E (IgE) levels in patients with eczema,and it is associated with the progression of immunoglobulin A nephropathy.Methods:The association between single nucleotide polymorphism (SNP) rs1800875,serum chymase,and serum IgE levels was examined in 175 CHD subjects and 95 non-CHD subjects.Results:Statistical analysis indicated no significant difference in allele frequency between CHD and non-CHD.However,a significant association was found between CMA1 genotypes and total IgE levels in CHD subjects.Meanwhile,crossover analysis revealed that,in GG homozygotes,CHD risk was nearly six times higher in those with IgE (U/ml) level <2.58 (natural logarithm conversion),while no association was found with chymase level.Conclusions:Polymorphism rs1800875 of CMA1 may be associated with serum IgE level in CHD subjects,but not with chymase level in both groups.In GG homozygotes,high IgE level is a protective factor against coronary disease.
