Hostname: page-component-586b7cd67f-r5fsc Total loading time: 0 Render date: 2024-11-26T07:58:53.307Z Has data issue: false hasContentIssue false

Streptococcus agalactiae-induced autophagy of bovine mammary epithelial cell via PI3K/AKT/mTOR pathway

Published online by Cambridge University Press:  07 April 2022

Mengzhu Qi
Affiliation:
College of Veterinary Medicine, Shandong Agricultural University, Tai`an, Shandong 271018, China
Hao Geng
Affiliation:
College of Veterinary Medicine, Shandong Agricultural University, Tai`an, Shandong 271018, China
Na Geng
Affiliation:
College of Veterinary Medicine, Shandong Agricultural University, Tai`an, Shandong 271018, China
Yukun Cui
Affiliation:
College of Veterinary Medicine, Shandong Agricultural University, Tai`an, Shandong 271018, China
Changxi Qi
Affiliation:
College of Veterinary Medicine, Shandong Agricultural University, Tai`an, Shandong 271018, China
Guodong Cheng
Affiliation:
College of Veterinary Medicine, Shandong Agricultural University, Tai`an, Shandong 271018, China
Kaimin Song
Affiliation:
College of Veterinary Medicine, Shandong Agricultural University, Tai`an, Shandong 271018, China
Liping Hu
Affiliation:
Shandong Provincial Center for Animal Disease Control and Prevention, Jinan, Shandong 251000, China
Yongxia Liu*
Affiliation:
College of Veterinary Medicine, Shandong Agricultural University, Tai`an, Shandong 271018, China Research Center for Animal Disease Control Engineering, Shandong Agricultural University, Tai`an, Shandong 271018, China
Jianzhu Liu*
Affiliation:
College of Veterinary Medicine, Shandong Agricultural University, Tai`an, Shandong 271018, China
Bo Han
Affiliation:
College of Veterinary Medicine, China Agricultural University, Beijing 100193, China
*
Authors for correspondence: Yongxia Liu, Email: [email protected] Jianzhu Liu, Email: [email protected]
Authors for correspondence: Yongxia Liu, Email: [email protected] Jianzhu Liu, Email: [email protected]

Abstract

Streptococcus agalactiae (S. agalactiae) infection is a significant cause of mastitis, resulting in loss of cellular homeostasis and tissue damage. Autophagy plays an essential function in cell survival, defense, and the preservation of cellular homeostasis, and is often part of the response to pathogenic challenge. However, the effect of autophagy induced by S. agalactiae in bovine mammary epithelial cells (bMECs) is mainly unknown. So in this study, an intracellular S. agalactiae infection model was established. Through evaluating the autophagy-related indicators, we observed that after S. agalactiae infection, a significant quantity of LC3-I was converted to LC3-II, p62 was degraded, and levels of Beclin1 and Bcl2 increased significantly in bMECs, indicating that S. agalactiae induced autophagy. The increase in levels of LAMP2 and LysoTracker Deep Red fluorescent spots indicated that lysosomes had participated in the degradation of autophagic contents. After autophagy was activated by rapamycin (Rapa), the amount of p-Akt and p-mTOR decreased significantly, whilst the amount of intracellular S. agalactiae increased significantly. Whereas the autophagy was inhibited by 3-methyladenine (3MA), the number of intracellular pathogens decreased. In conclusion, the results demonstrated that S. agalactiae could induce autophagy through PI3K/Akt/mTOR pathway and utilize autophagy to survive in bMECs.

Type
Research Article
Copyright
Copyright © The Author(s), 2022. Published by Cambridge University Press on behalf of Hannah Dairy Research Foundation

Access options

Get access to the full version of this content by using one of the access options below. (Log in options will check for institutional or personal access. Content may require purchase if you do not have access.)

References

Almeida, A, Alves-Barroco, C, Sauvage, E, Bexiga, R and Glaser, P (2016) Persistence of a dominant bovine lineage of group B Streptococcus reveals genomic signatures of host adaptation. Environmental Microbiology 18, 42164229.CrossRefGoogle ScholarPubMed
Babuta, M, Furi, I, Bala, S, Bukong, TN, Lowe, P, Catalano, D, Calenda, C, Kodys, K and Szabo, G (2019) Dysregulated autophagy and lysosome function are linked to exosome production by micro-RNA 155 in alcoholic liver disease. Hepatology 70, 21232141.CrossRefGoogle ScholarPubMed
Chikte, S, Panchal, N and Warnes, G (2014) Use of LysoTracker dyes: a flow cytometric study of autophagy. Cytometry Part A 85, 169178.CrossRefGoogle ScholarPubMed
De Gaetano, GV, Pietrocola, G, Romeo, L, Galbo, R and Beninati, C (2018) The Streptococcus agalactiae cell wall-anchored protein PbsP mediates adhesion to and invasion of epithelial cells by exploiting the host vitronectin/α v integrin axis. Molecular Microbiology 110, 8294.CrossRefGoogle ScholarPubMed
Eskelinen, EL (2006) Roles of LAMP-1 and LAMP-2 in lysosome biogenesis and autophagy. Molecular Aspects of Medicine 27, 495502.CrossRefGoogle ScholarPubMed
Feng, FB and Qiu, HY (2018) Effects of Artesunate on chondrocyte proliferation, apoptosis and autophagy through the PI3K/AKT/mTOR signaling pathway in rat models with rheumatoid arthritis. Biomedicine & Pharmacotherapy 102, 12091220.CrossRefGoogle ScholarPubMed
Fukuda, M, Asano, S, Nakamura, T, Adachi, M, Yoshida, M, Yanagida, M and Nishida, E (1997) CRM1 is responsible for intracellular transport mediated by the nuclear export signal. Nature 390, 308311.CrossRefGoogle ScholarPubMed
Geng, N, Liu, K, Lu, J, Xu, Y and Han, B (2020 a) Autophagy of bovine mammary epithelial cell induced by intracellular Staphylococcus aureus. The Journal of Microbiology 58, 320329.CrossRefGoogle ScholarPubMed
Geng, N, Wang, X, Yu, X, Wang, R and Liu, Y (2020 b) Staphylococcus aureus avoids autophagy clearance of bovine mammary epithelial cells by impairing lysosomal function. Frontiers in Immunology 11, 746.CrossRefGoogle ScholarPubMed
He, C, Bassik, MC, Moresi, V, Sun, K, Wei, Y, Zou, Z, An, Z, Loh, J, Fisher, J and Sun, Q (2012) Exercise-induced BCL2-regulated autophagy is required for muscle glucose homeostasis. Nature 481, 511515.CrossRefGoogle ScholarPubMed
Jiang, P and Mizushima, N (2015) LC3- and p62-based biochemical methods for the analysis of autophagy progression in mammalian cells. Methods (San Diego, Calif.) 75, 1318.CrossRefGoogle ScholarPubMed
Jørgensen, HJ, Nordstoga, A, Sviland, S, Zadoks, RN, Sølverød, L, Kvilte, B and Mørk, T (2015) Streptococcus agalactiae in the environment of bovine dairy herds – rewriting the textbooks? Veterinary Microbiology 184, 6472.CrossRefGoogle ScholarPubMed
Kang, R, Zeh, HJ, Lotze, MT and Tang, D (2011) The Beclin 1 network regulates autophagy and apoptosis. Cell Death & Differentiation 18, 571580.CrossRefGoogle ScholarPubMed
Liu, G, Pei, F, Yang, F, Li, L, Amit, A, Liu, S, Buchan, J and William, C (2017) Role of autophagy and apoptosis in non-small-cell lung cancer. International Journal of Molecular Sciences 18, 367.CrossRefGoogle ScholarPubMed
Lopiccolo, J, Blumenthal, GM, Bernstein, WB and Dennis, PA (2008) Targeting the PI3K/Akt/mTOR pathway: effective combinations and clinical considerations. Drug Resistance Updates 11, 3250.CrossRefGoogle ScholarPubMed
Lorenzini, A, Tresini, M, Mawal-Dewan, M, Frisoni, L, Hong, Z, Allen, RG, Sell, C and Cristofalo, VJ (2002) Role of the Raf/MEK/ERK and the PI3K/Akt(PKB) pathways in fibroblast senescence. Experimental Gerontology 37, 11491156.CrossRefGoogle ScholarPubMed
Lu, N, Li, X, Tan, R, An, J, Cai, Z, Hu, X, Wang, F, Wang, H, Lu, C and Lu, H (2018) HIF-1α/Beclin1-mediated autophagy is involved in neuroprotection induced by hypoxic preconditioning. Journal of Molecular Neuroscience 66, 238250.CrossRefGoogle ScholarPubMed
Maejima, Y, Kyoi, S, Zhai, P, Tong, L and Sadoshima, J (2013) Mst1 inhibits autophagy by promoting Beclin1-Bcl-2 interaction. Nature Medicine 19, 14781488.CrossRefGoogle Scholar
Martins, WK, Santos, NF, Rocha, CDS, Bacellar, IOL and Baptista, MS (2018) Parallel damage in mitochondria and lysosomes is an efficient way to photoinduce cell death. Autophagy 15, 259279.CrossRefGoogle ScholarPubMed
Mawal-Dewan, M, Lorenzini, A, Frisoni, L, Zhang, H, Cristofalo, VJ and Sell, C (2002) Regulation of collagenase expression during replicative senescence in human fibroblasts by Akt-forkhead signaling. Journal of Biological Chemistry 277, 78577864.CrossRefGoogle ScholarPubMed
Mukhopadhyay, S, Panda, PK, Sinha, N, Das, DN and Bhutia, SK (2014) Autophagy and apoptosis: where do they meet? Apoptosis 19, 555566.CrossRefGoogle ScholarPubMed
Niu, H, Zhang, H, Wu, F, Xiong, B, Tong, J and Jiang, L (2020) Proteomics study on the protective mechanism of soybean isoflavone against inflammation injury of bovine mammary epithelial cells induced by Streptococcus agalactiae. Cell Stress and Chaperones 26, 91101.CrossRefGoogle Scholar
Pang, M, Sun, L, He, T, Bao, H, Zhang, L, Zhou, Y, Zhang, H, Wei, R, Liu, Y and Wang, R (2017) Molecular and virulence characterization of highly prevalent Streptococcus agalactiae circulated in bovine dairy herds. Veterinary Research 48, 65.CrossRefGoogle ScholarPubMed
Pierzyńska-Mach, A, Janowski, PA and Dobrucki, JW (2014) Evaluation of acridine orange, LysoTracker Red, and quinacrine as fluorescent probes for long-term tracking of acidic vesicles. Cytometry Part A 85, 729737.CrossRefGoogle ScholarPubMed
Run, W, Wen, Z, Lumei, W, Na, G, Xiaozhou, W, Meihua, Z and Jianzhu, L, Yongxia, L and Bo, H (2021) Intracellular Staphylococcus aureus inhibits autophagy of bovine mammary epithelial cells through activating p38α. Journal of Dairy Research 88, 293301.Google Scholar
Runwal, G, Stamatakou, E, Siddiqi, FH, Puri, C and Rubinsztein, DC (2019) LC3-positive structures are prominent in autophagy-deficient cells. Scientific Reports 9, 10147.CrossRefGoogle ScholarPubMed
Seegers, H, Fourichon, C and Beaudeau, F (2003) Production effects related to mastitis and mastitis economics in dairy cattle herds. Veterinary Research 34, 475491.CrossRefGoogle ScholarPubMed
Tanaka, Y, Guhde, G, Suter, A, Eskelinen, EL, Hartmann, D, Lullmann-Rauch, R, Janssen, P, Blanz, J, Figura, KV and Saftig, P (2000) Accumulation of autophagic vacuoles and cardiomyopathy in LAMP-2-deficient mice. Nature 406, 902906.CrossRefGoogle ScholarPubMed
Tong, J, Sun, M, Zhang, H, Yang, D and Jiang, L (2020) Proteomic analysis of bovine mammary epithelial cells after in vitro incubation with S. agalactiae: potential biomarkers. Veterinary Research 51, 98.CrossRefGoogle Scholar
Tu, QD, Jin, J, Hu, X, Ren, Y, Zhao, L and He, Q (2020) Curcumin improves the renal autophagy in rat experimental membranous nephropathy via regulating the PI3K/AKT/mTOR and Nrf2/HO-1 signaling pathways. Biomed Research International 2020, 7069052.Google ScholarPubMed
Yang, J, Pi, C and Wang, G (2018) Inhibition of PI3K/Akt/mTOR pathway by apigenin induces apoptosis and autophagy in hepatocellular carcinoma cells. Biomedicine & Pharmacotherapy 103, 699707.CrossRefGoogle ScholarPubMed
Zhang, A, Song, Y, Zhang, Z, Jiang, S and Ni, G (2021) Effects of autophagy inhibitor 3-Methyladenine on ischemic stroke: a protocol for systematic review and meta-analysis. Medicine 100, e23873.CrossRefGoogle ScholarPubMed
Zhao, Z, Sun, C, Chen, L, Qin, J and Li, W (2019) Inorganic nitrite increases the susceptibility of tilapia (Oreochromis niloticus) leucocytes to Streptococcus agalactiae. Fish & Shellfish Immunology 97, 111.CrossRefGoogle ScholarPubMed
Zhitomirsky, B, Farber, H and Assaraf, YG (2018) LysoTracker and MitoTracker Red are transport substrates of P-glycoprotein: implications for anticancer drug design evading multidrug resistance. Journal of Cellular & Molecular Medicine 22, 21312141.CrossRefGoogle ScholarPubMed