金黄色葡萄球菌对奶牛乳腺成纤维细胞TGF-β1/Smad信号通路及其转分化的影响

doi:10.11843/j.issn.0366-6964.2016.07.025

Abstract

Abstract:

This study aimed to investigate the effect of Staphylococcus aureus (S.aureus) on TGF-β1/Smad signaling pathway and transdifferentiation in bovine mammary fibroblasts (BMFB).BMFB were stimulated with heat-killed S.aureus (HKSA) at 10⁴，10⁵，10⁶，10⁷，10⁸ CFU•mL^-1.The transcription of TGF-β1，α-SMA and collagen-Ⅰ mRNA were detected by Real-time PCR after 24 h.The expression of α-SMA，collagen-Ⅰ and p-Smad2/3 protein were detected by Western blot.BMFB were pretreated with TGF-β1 receptor-specific inhibitor SB-431542，BMFB were stimulated with heat-killed S.aureus (HKSA) at 105 CFU•mL^-1.The transcription of α-SMA and collagen-Ⅰ mRNA were detected by Real-time PCR after 24 h.The expression of α-SMA，collagen-Ⅰ and p-Smad2/3 protein were detected by Western blot.Immunofluorescence was used to detect collagen-Ⅰ expression.The results showed that the transcription level of TGF-β1 mRNA，expression of α-SMA，collagen-Ⅰ and p-Smad2/3 increased significantly at different concentration after 24 h stimulation compared with unstimulated BMFB (P<0.05).The expression of TGF-β1，α-SMA，collagen-Ⅰ and p-Smad2/3 reached the maximal level at 105 CFU•mL^-1.The expression of α-SMA，collagen-Ⅰ and p-Smad2/3 were significantly reduced by pretreating BMFB with SB-431542 (P<0.01).The results indicate that TGF-β1/Smad signaling pathway plays an important role in S.aureus induced BMFB transdifferentiation.

Key words: Staphylococcus aureus, bovine mammary fibroblasts, α-SMA, collagen-Ⅰ, p-Smad2/3

CLC Number:

S852.3

YANG Bin，XU Dan-dan，SUN Zhi-peng，ZHAO Jia-qi，YAN Bo-wei，LI Xiao-ting，WU Rui. The Effect of Staphylococcus aureus on TGF-β1/Smad Signaling Pathway and Transdifferentiation in Bovine Mammary Fibroblasts[J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2016, 47(7): 1495-1501.

References

［1］BAR-GAL G K，BLUM S E，HADAS L，et al.Host-specificity of Staphylococcus aureus causing intramammary infections in dairy animals assessed by genotyping and virulence genes［J］.Vet Microbiol，2015，176(1-2)：143-154.
［2］AKERS R M，NICKERSON S C.Mastitis and its impact on structure and function in the ruminant mammary gland［J］.J Mammary Gland Biol Neoplasia，2011，16(4)：275-289.
［3］MINTON K.Extracellular matrix：Preconditioning the ECM for fibrosis［J］.Nat Rev Mol Cell Biol，2014，15(12)：766-767.
［4］RITTIÉ L.Another dimension to the importance of the extracellular matrix in fibrosis［J］. J Cell Commun Signal，2015，9(1)：99-100.
［5］BONNANS C，CHOU J，WERB Z.Remodelling the extracellular matrix in development and disease［J］.Nat Rev Mol Cell Biol，2014，15(12)：786-801.
［6］HINZ B.Myofibroblasts［J］.Exp Eye Res，2016，142：56-70.
［7］KENDALL R T，FEGHALI-BOSTWICK C A.Fibroblasts in fibrosis：novel roles and mediators［J］.Front Pharmacol，2014，5：123.
［8］BIERNACKA A，DOBACZEWSKI M，FRANGOGIANNIS N G.TGF-β signaling in fibrosis［J］.Growth Factors，2011，29(5)：196-202.
［9］NAKERAKANTI S，TROJANOWSKA M.The role of TGF-β receptors in fibrosis［J］.Open Rheumatol J，2012，6：156-162.
［10］MENG X M，TANG P M，LI J，et al.TGF-β/Smad signaling in renal fibrosis［J］.Front Physiol，2015，6：82.
［11］ANDREOTTI C S，PEREYRA E A，BARAVALLE C，et al.Staphylococcus aureus chronic intramammary infection modifies protein expression of transforming growth factor β (TGF-β) subfamily components during active involution［J］. Res Vet Sci，2014，96(1)：5-14.
［12］WANG X，XIU L，HU Q，et al.Deep sequencing-based transcriptional analysis of bovine mammary epithelial cells gene expression in response to in vitro infection with Staphylococcus aureus stains［J］. PLoS One，2013，8(12)：e82117.
［13］FU Y，ZHOU E，LIU Z，et al.Staphylococcus aureus and Escherichia coli elicit different innate immune responses from bovine mammary epithelial cells［J］.Vet Immunol Immunopathol，2013，155(4)：245-252.
［14］DARBY I A，LAVERDET B，BONTÉ F，et al.Fibroblasts and myofibroblasts in wound healing［J］.Clin Cosmet Investig Dermatol，2014，7：301-311.
［15］丁旭娜.TGF-β1、bFGF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.
DING X N.Studies on the effect of TGF-β1，bFGF and PDGF in bovine mammary fibrosis［D］.Huhhot：Inner Mongolia Agricultural University，2012.(in Chinese)
［16］YOSHIDA K，MURATA M，YAMAGUCHI T，et al.TGF-β/Smad signaling during hepatic fibro-carcinogenesis (review)［J］.Int J Oncol，2014，45(4)：1363-1371.
［17］LAN H Y.Diverse roles of TGF-β/Smads in renal fibrosis and inflammation［J］.Int J Biol Sci，2011，7(7)：1056-1067.
［18］FUJITA M，SHANNON J M，MORIKAWA O，et al.Overexpression of tumor necrosis factor-alpha diminishes pulmonary fibrosis induced by bleomycin or transforming growth factor-beta［J］.Am J Respir Cell Mol Biol，2003，29(6)：669-676.
［19］LAW B A，CARVER W E.Activation of cardiac fibroblasts by ethanol is blocked by TGF-β inhibition［J］.Alcohol Clin Exp Res，2013，37(8)：1286-1294.
［20］XIE L，ZHOU D，XIONG J，et al.Paraquat induce pulmonary epithelial-mesenchymal transition through transforming growth factor-β1-dependent mechanism［J］. Exp Toxicol Pathol，2016，68(1)：69-76.

[1]	GONG Yongping, CHEN Zhenrong, YIN Wenqi, WEN Jifeng, YI Keke, YAN Qigui. Analysis of Multidrug-resistant Genomic Island PRI1 of Enterococcus faecium LS170308 from Moschus berezovskii [J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2019, 50(6): 1275-1283.
[2]	GONG Yongping, CHEN Zhenrong, YIN Wenqi, WEN Jifeng, YI Keke, YAN Qigui. Analysis of Multidrug-resistant Genomic Island PRI1 of Enterococcus faecium LS170308 from Moschus berezovskii [J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2019, 50(6): 1275-1283.
[3]	ZHANG Hang, YANG Feng, LI Xinpu, WANG Xurong, LUO Jinyin, LI Hongsheng. Study on Relationship between Antimicrobial Resistance and Efflux Pump Related Genes of Methicillin-Resistant Staphylococcus aureus in Dairy Cow [J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2019, 50(11): 2326-2332.
[4]	MA Weiwu, ZHOU Xuezhang. Expression of Panton-Valenine Leukocidin from Staphylococcus aureus and Its Effects on Bovine Mammary Epithelial Cells [J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2019, 50(10): 2105-2112.
[5]	CHEN Yao, DING Hong-lei, HE Ying, MU Hao, WANG Li-min, WANG Hao-ju. Antimicrobial Susceptibility Analysis and ESBL Gene Detection of Staphylococcus aureus Isolated at Animal Farms in Chongqing [J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2018, 49(5): 1074-1080.
[6]	CHEN Yao, DING Hong-lei, HE Ying, MU Hao, WANG Li-min, WANG Hao-ju. Antimicrobial Susceptibility Analysis and ESBL Gene Detection of Staphylococcus aureus Isolated at Animal Farms in Chongqing [J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2018, 49(5): 1074-1080.
[7]	LIU Gui-lan, ZHOU Yong-lin, LI Wen-hua, XIE Ling-ling, LIU Ai-ling, DENG Xu-ming. The Antimicrobial and Anti-hemolysis Effects of Total Magnolol on Staphylococcus aureus [J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2018, 49(2): 432-438.
[8]	LIU Gui-lan, ZHOU Yong-lin, LI Wen-hua, XIE Ling-ling, LIU Ai-ling, DENG Xu-ming. The Antimicrobial and Anti-hemolysis Effects of Total Magnolol on Staphylococcus aureus [J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2018, 49(2): 432-438.
[9]	MENG Dan, MENG Qing-ling, QIAO Jun, CAI Kuo-jun, WANG Deng-feng, MA Shuai, WU Ye-hui, CAI Xue-peng. Study on Resistance Characteristics, Virulence Genes and Molecular Classification of Staphylococcus aureus Isolated from Cows in Xinjiang [J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2018, 49(1): 181-194.
[10]	MENG Dan, MENG Qing-ling, QIAO Jun, CAI Kuo-jun, WANG Deng-feng, MA Shuai, WU Ye-hui, CAI Xue-peng. Study on Resistance Characteristics, Virulence Genes and Molecular Classification of Staphylococcus aureus Isolated from Cows in Xinjiang [J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2018, 49(1): 181-194.
[11]	LI Yu-hua, FU Jie-li, LI Pei-fei, YANG Qiang-zhen, WANG Li-rui, XIE Wei-yi, LI Xin-hong. Effects of Staphylococcus aureus on Motility and Protein Phosphorylation Modification of Boar Sperm during Storage in vitro [J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2017, 48(9): 1665-1673.
[12]	LI Yu-hua, FU Jie-li, LI Pei-fei, YANG Qiang-zhen, WANG Li-rui, XIE Wei-yi, LI Xin-hong. Effects of Staphylococcus aureus on Motility and Protein Phosphorylation Modification of Boar Sperm during Storage in vitro [J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2017, 48(9): 1665-1673.
[13]	XU Guo-yang, FU Li-zhi, YANG Jin-long, YANG Liu,XU Deng-feng, ZHANG Yi-fan, SHEN Ke-fei, ZHANG Su-hui. Isolation and Identification of Pathogenic Bacteria of Lymphadenitis in Goat and Establishment of Multiplex PCR Detection Method [J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2017, 48(2): 324-330.
[14]	XU Guo-yang, FU Li-zhi, YANG Jin-long, YANG Liu,XU Deng-feng, ZHANG Yi-fan, SHEN Ke-fei, ZHANG Su-hui. Isolation and Identification of Pathogenic Bacteria of Lymphadenitis in Goat and Establishment of Multiplex PCR Detection Method [J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2017, 48(2): 324-330.
[15]	YANG Bin，XU Dan-dan，SUN Zhi-peng，ZHAO Jia-qi，YAN Bo-wei，LI Xiao-ting，WU Rui. The Effect of Staphylococcus aureus on TGF-β1/Smad Signaling Pathway and Transdifferentiation in Bovine Mammary Fibroblasts [J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2016, 47(7): 1495-1501.

The Effect of Staphylococcus aureus on TGF-β1/Smad Signaling Pathway and Transdifferentiation in Bovine Mammary Fibroblasts

PDF (PC)

Knowledge

Abstract

Cite this article

share this article

References

Related Articles 15

Recommended Articles

Metrics

Comments