奶牛乳腺炎金黄色葡萄球菌诱导小鼠乳腺组织中TβRⅠ、FGFR2和PDGFRβ的表达
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摘要
为探究TβRⅠ、FGFR2和PDGFRβ对小鼠乳腺组织纤维化的作用,本研究采用奶牛乳腺炎金黄色葡萄球菌(S.aureus)感染小鼠乳腺后,分别于感染后1 d、3 d、5 d、7 d、14 d、21 d采集小鼠乳腺组织,应用免疫组化染色检测TβRⅠ、FGFR2和PDGFRβ在小鼠乳腺组织细胞中的表达与分布,应用荧光定量PCR和western blot方法检测小鼠乳腺组织中TβRⅠ、FGFR2和PDGFRβm RNA转录水平和这些蛋白的相对表达水平。结果显示,小鼠感染S.aureus前期(1 d~5 d) TβRⅠ、FGFR2和PDGFRβ主要在其炎性细胞和乳腺上皮细胞中表达,TβRⅠ、FGFR2和PDGFRβm RNA转录水平和蛋白表达极显著高于对照组(p<0.001),感染S.aureus后期(7 d~21 d),细胞外基质较多,TβRⅠ、FGFR2和PDGFRβ在小鼠乳腺成纤维细胞和乳腺上皮细胞中均有表达,TβRⅠ、FGFR2和PDGFRβ的表达极显著高于对照组(p<0.001)。研究表明,S.aureus感染小鼠后,能够促进乳腺组织中TβRⅠ、FGFR2和PDGFRβ的表达,从而促进小鼠乳腺纤维化。本研究为深入探究生长因子受体在乳腺纤维化的分子作用机制奠定了基础。
To study the relationship of TβR Ⅰ, FGFR2 and PDGFRβ expression to fibrosis in mice mammary gland tissue,mammary gland tissue of mice was collected after 1 d, 3 d, 5 d, 7 d, 14 d and 21 d post Staphylococcus aureus-infection. The expressions of TβRⅠ, FGFR2 and PDGFRβ in mammary gland tissue were detected by immunohistochemical staining, real time PCR and western blot. The results showed that TβRⅠ, FGFR2 and PDGFRβ were mainly expressed in mammary fibroblasts from1 d to 5 d, then in both mammary fibroblasts and mammary epithelial cells from 7 d to 21 d. Both m RNA and protein expressions of TβRⅠ, FGFR2 and PDGFRβ were significantly up-regulated when mice were infected with S.aureus. These results indicated the expressions of TβRⅠ, FGFR2 and PDGFRβ in mice mammary gland tissue were induced by S.aureus.
To study the relationship of TβR Ⅰ, FGFR2 and PDGFRβ expression to fibrosis in mice mammary gland tissue,mammary gland tissue of mice was collected after 1 d, 3 d, 5 d, 7 d, 14 d and 21 d post Staphylococcus aureus-infection. The expressions of TβRⅠ, FGFR2 and PDGFRβ in mammary gland tissue were detected by immunohistochemical staining, real time PCR and western blot. The results showed that TβRⅠ, FGFR2 and PDGFRβ were mainly expressed in mammary fibroblasts from1 d to 5 d, then in both mammary fibroblasts and mammary epithelial cells from 7 d to 21 d. Both m RNA and protein expressions of TβRⅠ, FGFR2 and PDGFRβ were significantly up-regulated when mice were infected with S.aureus. These results indicated the expressions of TβRⅠ, FGFR2 and PDGFRβ in mice mammary gland tissue were induced by S.aureus.
引文
[1]Zhao Xin,Lacasse P.Mammary tissue damage during bovine mastitis:causes and control[J].J Anim Sci,2008,86(13):57-65.
[2]Hillerton J E,Berry E A.Treating mastitis in the cow-a tradition or an archaism[J].J Appl Microbiol,2005,98(6):1250-1255.
[3]丁旭娜.TGF-β1、b FGF和PDGF在奶牛乳腺纤维化中作用的研究[D]:呼和浩特:内蒙古农业大学,2012.
[4]Ahn J Y,Kim M H,Lim M J,et al.The inhibitory effect of ginsan on TGF-βmediated fibrotic process[J].J Cellul Physiol,2011,226(5):1241-1247.
[5]Li Xiao-xi,Chang Hong-en,Nai Wen-qing,et al.Pulmonary expression levels of fibroblast growth factor receptors and lung fibrosis in mice at different ages[J].Nan Fang Yi Ke Da Xue Xue Bao,2013,33(4):607-610.
[6]Fan Bin,Ma Li-kun,Li Qian,et al.Role of PDGFs/PDGFRs signaling pathway in myocardial fibrosis of DOCA/salt hypertensive rats[J].Int Clin Exp Pathol,2014,7(1):16-27.
[7]Raimondi A R,Molinolo A A,Itoiz M E.Fibroblast growth factor-2 expression during experimental oral carcinogenesis.Its possible role in the induction of pre-malignant fibrosis[J].JOral Pathol Med,2010,35(4):212-217.
[8]Daoussis D,Tsamandas A C,Liossis S N C,et al.B-cell depletion therapy in patients with diffuse systemic sclerosis associates with a significant decrease in PDGFR expression and activation in spindle-like cells in the skin[J].Arthritis Res Ther,2012,14(3):R145.
[9]Choi M E.Mechanism of transforming growth factor-beta1 signaling[J].Kidney Int,2000,58(77):S53-S58.
[10]毕艳楠,王凤龙,丁玉林,等.金黄色葡萄球菌诱导小鼠乳腺纤维化模型TGF-β1的表达[J].中国预防兽医学报,2018,40(3):238-242.
[11]刘珍.呼和浩特地区临床型奶牛乳腺炎病原分离鉴定及其对小白鼠致病性的研究[D]:呼和浩特:内蒙古农业大学,2005.
[12]赵素微.奶牛乳腺炎主要病原菌致病性及其免疫原性研究[D]:呼和浩特:内蒙古农业大学,2008.
[13]Kiku Y,Nagasawa Y,Tanabe F,et al.The cell wall component lipoteichoic acid of Staphylococcus aureus induces chemokine gene expression in bovine mammary epithelialcells[J].Vet Med Sci,2016,78(9):1505-1510.
[14]Castilho I G,Sta D,Langoni H,et al.Host-pathogen interactions in bovine mammary epithelial cells and HeL a cells by Staphylococcus aureus isolated from subclinical bovine mastitis[J].J Dairy Sci,2017,100(8):6414-6421.
[15]吴建美.金黄色葡萄球菌对奶牛乳腺细胞几种生长因子表达及其信号机制作用的研究[D]:呼和浩特:内蒙古农业大学,2015.
[16]Lehtonen S T,Veijola A,Karvonen H,et al.Pirfenidone and nintedanib modulate properties of fibroblasts and myofibroblasts in idiopathic pulmonary fibrosis[J].Resp Res,2016,17(1):14.
[17]李娟,郑全辉,郝小惠,等.转化生长因子β1及其受体TβRⅠ、TβRⅡ在矽尘致大鼠肺纤维化中表达[J].中国职业医学,2013,40(4):305-308.
[18]Mackenzie B A,Korfei M,Henneke I,et al.Increased FGF1-FGFRc expression in idiopathic pulmonary fibrosis[J].Resp Res,2015,16(1):83.
[19]Hes G M,Tzankov A,Hebeda K,et al.Platelet derived growth factor receptorβ(PDGFRβ)immunohistochemistry highlights activated bone marrow stroma and is potentially predictive for fibrosis progression in prefibrotic myeloproliferative neoplasia[J].Histopathol,2015,67(5):617-624.
[20]江宇杰,张燕,石小枫,等.TIMP1/2和TGF-β1siRNA对肝纤维化大鼠PDGF/ERK信号通路的影响[D].重庆:重庆医科大学,2014.
[21]刘印,贺利平,田京.不同时期b FGF或副甲状腺激素相关肽对TGF-β1诱导的兔BMSCs向软骨细胞分化的影响[J].中国修复重建外科杂志,2013,27(2):199-202.
[22]Wu Jian-mei,Ding Yu-lin,Bi Yan-nan,et al.Staphylococcus aureus induces TGF-β1 and bFGF expression through the activation of AP-1 and NF-κB transcription factors in bovine mammary gland fibroblasts[J].Microb Pathog,2016,95:7-14.
[2]Hillerton J E,Berry E A.Treating mastitis in the cow-a tradition or an archaism[J].J Appl Microbiol,2005,98(6):1250-1255.
[3]丁旭娜.TGF-β1、b FGF和PDGF在奶牛乳腺纤维化中作用的研究[D]:呼和浩特:内蒙古农业大学,2012.
[4]Ahn J Y,Kim M H,Lim M J,et al.The inhibitory effect of ginsan on TGF-βmediated fibrotic process[J].J Cellul Physiol,2011,226(5):1241-1247.
[5]Li Xiao-xi,Chang Hong-en,Nai Wen-qing,et al.Pulmonary expression levels of fibroblast growth factor receptors and lung fibrosis in mice at different ages[J].Nan Fang Yi Ke Da Xue Xue Bao,2013,33(4):607-610.
[6]Fan Bin,Ma Li-kun,Li Qian,et al.Role of PDGFs/PDGFRs signaling pathway in myocardial fibrosis of DOCA/salt hypertensive rats[J].Int Clin Exp Pathol,2014,7(1):16-27.
[7]Raimondi A R,Molinolo A A,Itoiz M E.Fibroblast growth factor-2 expression during experimental oral carcinogenesis.Its possible role in the induction of pre-malignant fibrosis[J].JOral Pathol Med,2010,35(4):212-217.
[8]Daoussis D,Tsamandas A C,Liossis S N C,et al.B-cell depletion therapy in patients with diffuse systemic sclerosis associates with a significant decrease in PDGFR expression and activation in spindle-like cells in the skin[J].Arthritis Res Ther,2012,14(3):R145.
[9]Choi M E.Mechanism of transforming growth factor-beta1 signaling[J].Kidney Int,2000,58(77):S53-S58.
[10]毕艳楠,王凤龙,丁玉林,等.金黄色葡萄球菌诱导小鼠乳腺纤维化模型TGF-β1的表达[J].中国预防兽医学报,2018,40(3):238-242.
[11]刘珍.呼和浩特地区临床型奶牛乳腺炎病原分离鉴定及其对小白鼠致病性的研究[D]:呼和浩特:内蒙古农业大学,2005.
[12]赵素微.奶牛乳腺炎主要病原菌致病性及其免疫原性研究[D]:呼和浩特:内蒙古农业大学,2008.
[13]Kiku Y,Nagasawa Y,Tanabe F,et al.The cell wall component lipoteichoic acid of Staphylococcus aureus induces chemokine gene expression in bovine mammary epithelialcells[J].Vet Med Sci,2016,78(9):1505-1510.
[14]Castilho I G,Sta D,Langoni H,et al.Host-pathogen interactions in bovine mammary epithelial cells and HeL a cells by Staphylococcus aureus isolated from subclinical bovine mastitis[J].J Dairy Sci,2017,100(8):6414-6421.
[15]吴建美.金黄色葡萄球菌对奶牛乳腺细胞几种生长因子表达及其信号机制作用的研究[D]:呼和浩特:内蒙古农业大学,2015.
[16]Lehtonen S T,Veijola A,Karvonen H,et al.Pirfenidone and nintedanib modulate properties of fibroblasts and myofibroblasts in idiopathic pulmonary fibrosis[J].Resp Res,2016,17(1):14.
[17]李娟,郑全辉,郝小惠,等.转化生长因子β1及其受体TβRⅠ、TβRⅡ在矽尘致大鼠肺纤维化中表达[J].中国职业医学,2013,40(4):305-308.
[18]Mackenzie B A,Korfei M,Henneke I,et al.Increased FGF1-FGFRc expression in idiopathic pulmonary fibrosis[J].Resp Res,2015,16(1):83.
[19]Hes G M,Tzankov A,Hebeda K,et al.Platelet derived growth factor receptorβ(PDGFRβ)immunohistochemistry highlights activated bone marrow stroma and is potentially predictive for fibrosis progression in prefibrotic myeloproliferative neoplasia[J].Histopathol,2015,67(5):617-624.
[20]江宇杰,张燕,石小枫,等.TIMP1/2和TGF-β1siRNA对肝纤维化大鼠PDGF/ERK信号通路的影响[D].重庆:重庆医科大学,2014.
[21]刘印,贺利平,田京.不同时期b FGF或副甲状腺激素相关肽对TGF-β1诱导的兔BMSCs向软骨细胞分化的影响[J].中国修复重建外科杂志,2013,27(2):199-202.
[22]Wu Jian-mei,Ding Yu-lin,Bi Yan-nan,et al.Staphylococcus aureus induces TGF-β1 and bFGF expression through the activation of AP-1 and NF-κB transcription factors in bovine mammary gland fibroblasts[J].Microb Pathog,2016,95:7-14.