Skip to main content

Advertisement

SpringerLink
Log in

Resveratrol inhibits LPS-induced apoptosis in bovine mammary epithelial cells: the role of PGC1α-SIRT3 axis

  • Published:
In Vitro Cellular & Developmental Biology - Animal Aims and scope Submit manuscript

Abstract

Resveratrol (Res) is a bioactive dietary component and alleviates apoptosis in multiple cell types. However, its effect and mechanism on lipopolysaccharide (LPS)-induced bovine mammary epithelial cells (BMEC) apoptosis, which commonly happens in dairy cows with mastitis, is unknown. We hypothesized that Res would inhibit LPS-induced apoptosis in BMEC through SIRT3, a NAD + -dependent deacetylase activated by Res. To test the dose–response effect on apoptosis, 0–50 μM Res were incubated with BMEC for 12 h, followed by 250 μg/mL LPS treatment for 12 h. To investigate the role of SIRT3 in Res-mediated alleviation of apoptosis, BMEC were pretreated with 50 μM Res for 12 h, then incubated with si-SIRT3 for 12 h and were finally treated with 250 μg/mL LPS for 12 h. Res dose-dependently promoted the cell viability and protein levels of Bcl-2 (Linear P < 0.001) but decreased protein levels of Bax, Caspase-3 and Bax/Bcl-2 (Linear P < 0.001). TUNEL assays indicated that cellular fluorescence intensity declined with the rising doses of Res. Res also dose-dependently upregulated SIRT3 expression, but LPS had the opposite effect. SIRT3 silencing abolished these results with Res incubation. Mechanically, Res enhanced the nuclear translocation of PGC1α, the transcriptional cofactor for SIRT3. Further molecular docking analysis revealed that Res could directly bind to PGC1α by forming a hydrogen bond with Tyr-722. Overall, our data suggested that Res relieved LPS-induced BMEC apoptosis through the PGC1α-SIRT3 axis, providing a basis for further in vivo investigations of applying Res to relieve mastitis in dairy cows.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Figure 1.
Figure 2.
Figure 3.
Figure 4.
Figure 5.
Figure 6.
Figure 7.
Figure 8.
Figure 9.

Similar content being viewed by others

Data availability

The original contributions presented in the study are included in the article, further inquiries can be directed to the corresponding author/s.

Abbreviations

BMEC :

Bovine mammary epithelial cells

Res :

Resveratrol

LPS :

Lipopolysaccharide

SIRT3 or Sirtuin3 :

Silent information regulator 2-like protein 3

E. coli :

Escherichia coli

CV :

Coefficient of variation

HRP :

Horseradish peroxidase

CCK-8 :

CCK-8 cell proliferation and cytotoxicity assay kit

TUNEL :

Terminal- deoxynucleoitidyl transferase mediated nick end labeling

WB :

Western-blotting

References

  • Baarine M, Thandapilly SJ, Louis XL, Mazue F, Liping Yu, Delmas D, Netticadan T, Lizard G, Latruffe N (2011) Pro-apoptotic versus anti-apoptotic properties of dietary resveratrol on tumoral and normal cardiac cells. Genes Nutr 6:161–169

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Bastin J, Djouadi F (2016) Resveratrol and myopathy. Nutrients 8:254

    Article  PubMed  PubMed Central  Google Scholar 

  • Bhandari S, Subedi D, Tiwari BB, Shrestha P, Shah S, Al-Mustapha AI (2021) Prevalence and risk factors for multidrug-resistant Escherichia coli isolated from subclinical mastitis in the western Chitwan region of Nepal. J Dairy Sci 104:12765–12772

    Article  CAS  PubMed  Google Scholar 

  • Cui B, Wang Y, Jin J, Yang Z, Guo R, Li X, Yang L, Li Z (2022) Resveratrol treats UVB-induced photoaging by anti-MMP expression, through anti-inflammatory, antioxidant, and antiapoptotic properties, and treats photoaging by upregulating VEGF-B expression. Oxid Med Cell Longev 2022:6037303

    Article  PubMed  PubMed Central  Google Scholar 

  • Dalanezi FM, Joaquim SF, Guimaraes FF, Guerra ST, Lopes BC, Schmidt EMS, Cerri RLA, Langoni H (2020) Influence of pathogens causing clinical mastitis on reproductive variables of dairy cows. J Dairy Sci 103:3648–3655

    Article  CAS  PubMed  Google Scholar 

  • Fujita Y, Yamashita T (2018) Sirtuins in neuroendocrine regulation and neurological diseases. Front Neurosci 12:778

    Article  PubMed  PubMed Central  Google Scholar 

  • Gao X, Dai J, Li G, Dai X (2021) Gambogic acid protects LPS-induced apoptosis and inflammation in a cell model of neonatal pneumonia through the regulation of TrkA/Akt signaling pathway. BMC Pharmacol Toxicol 22:28

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • He W, Ma S, Lei L, He J, Li X, Tao J, Wang X, Song S, Wang Y, Wang Y, Shen J, Cai C, Congming Wu (2020) Prevalence, etiology, and economic impact of clinical mastitis on large dairy farms in China. Vet Microbiol 242:108570

    Article  PubMed  Google Scholar 

  • Hou X, Yang S, Yin J (2019) Blocking the REDD1/TXNIP axis ameliorates LPS-induced vascular endothelial cell injury through repressing oxidative stress and apoptosis. Am J Physiol Cell Physiol 316:C104–C110

    Article  CAS  PubMed  Google Scholar 

  • Hu L-F, Lan H-R, Li X-M, Jin K-T (2021) A systematic review of the potential chemoprotective effects of resveratrol on doxorubicin-induced cardiotoxicity: focus on the antioxidant, antiapoptotic, and anti-inflammatory activities. Oxid Med Cell Longev 2021:2951697

    Article  PubMed  PubMed Central  Google Scholar 

  • Lasa A, Churruca I, Eseberri I, Andres-Lacueva C, Portillo MP (2012) Delipidating effect of resveratrol metabolites in 3T3-L1 adipocytes. Mol Nutr Food Res 56:1559–1568

    Article  CAS  PubMed  Google Scholar 

  • Lee HJ, Kang M-G, Cha HY, Kim YM, Lim Y, Yang SJ (2019) Effects of piceatannol and resveratrol on sirtuins and hepatic inflammation in high-fat diet-fed mice. J Med Food 22:833–840

    Article  CAS  PubMed  Google Scholar 

  • Liu M, Fang G, Yin S, Zhao X, Zhang C, Li J, Liu Z (2019) Caffeic acid prevented LPS-induced injury of primary bovine mammary epithelial cells through inhibiting NF-kappaB and MAPK activation. Mediators Inflamm 2019:1897820

    Article  PubMed  PubMed Central  Google Scholar 

  • Liu M, Zhang C, Xiaolong Xu, Zhao X, Han Z, Liu D, Bo R, Li J, Liu Z (2021) Ferulic acid inhibits LPS-induced apoptosis in bovine mammary epithelial cells by regulating the NF-κB and Nrf2 signalling pathways to restore mitochondrial dynamics and ROS generation. Vet Res 52:104

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Ma Z, Wang D, Weng J, Zhang S, Zhang Y (2020) BNIP3 decreases the LPS-induced inflammation and apoptosis of chondrocytes by promoting the development of autophagy. J Orthop Surg Res 15:284

    Article  PubMed  PubMed Central  Google Scholar 

  • Malaguarnera L (2019) Influence of resveratrol on the immune response. Nutrients 11:946

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Malcata FB, Theo Pepler P, O’Reilly EL, Nicola Brady P, Eckersall D, Zadoks RN, Viora L (2020) Point-of-care tests for bovine clinical mastitis: what do we have and what do we need? J Dairy Res 87:60–66

    Article  CAS  PubMed  Google Scholar 

  • Matsushima S, Sadoshima J (2015) The role of sirtuins in cardiac disease. Am J Physiol Heart Circ Physiol 309:H1375–H1389

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • McNeel AK, Reiter BC, Weigel D, Osterstock J, Di Croce FA (2017) Validation of genomic predictions for wellness traits in US Holstein cows. J Dairy Sci 100:9115–9124

    Article  CAS  PubMed  Google Scholar 

  • Sergi D, Gelinas A, Beaulieu J, Renaud J, Tardif-Pellerin E, Guillard J, Martinoli MG (2021) Anti-apoptotic and anti-inflammatory role of trans epsilon-viniferin in a neuron-glia co-culture cellular model of Parkinson’s disease. Foods 10:586

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Steele NM, Swartz TH, Enger KM, Schramm H, Cockrum RR, Lacy-Hulbert SJ, White RR, Hogan J, Petersson-Wolfe CS (2019) The effect of J5 bacterins on clinical, behavioral, and antibody response following an Escherichia coli intramammary challenge in dairy cows at peak lactation. J Dairy Sci 102:11233–11249

    Article  CAS  PubMed  Google Scholar 

  • Sundaresan NR, Samant SA, Pillai VB, Rajamohan SB, Gupta MP (2008) SIRT3 is a stress-responsive deacetylase in cardiomyocytes that protects cells from stress-mediated cell death by deacetylation of Ku70. Mol Cell Biol 28:6384–6401

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Timmers S, Konings E, Bilet L, Houtkooper RH, van de Weijer T, Goossens GH, Hoeks J, van der Krieken S, Ryu D, Kersten S, Moonen-Kornips E, Hesselink MKC, Kunz I, Schrauwen-Hinderling VB, Blaak E, Auwerx J, Schrauwen P (2011) Calorie restriction-like effects of 30 days of resveratrol supplementation on energy metabolism and metabolic profile in obese humans. Cell Metab 14:612–622

    Article  CAS  PubMed  Google Scholar 

  • Wang H, Guan Y, Karamercan MA, Ye L, Bhatti T, Becker LB, Baur JA, Sims CA (2015) Resveratrol rescues kidney mitochondrial function following hemorrhagic shock. Shock 44:173–180

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Wang M, Zhu CQ, Zeng L, Cheng L, Ma L, Zhang M, Zhang YZ (2021) Melatonin regulates the cross-talk between autophagy and apoptosis by SIRT3 in testicular Leydig cells. Biochem Biophys Res Commun 555:182–189

    Article  CAS  PubMed  Google Scholar 

  • Wang YY, Sun SP, Zhu HS, Jiao XQ, Zhong K, Guo YJ, Zha GM, Han LQ, Yang GY, Li HP (2018) GABA regulates the proliferation and apoptosis of MAC-T cells through the LPS-induced TLR4 signaling pathway. Res Vet Sci 118:395–402

    Article  CAS  PubMed  Google Scholar 

  • Wang Z, Sun R, Wang G, Chen Z, Li Y, Zhao Y, Liu D, Zhao H, Zhang F, Yao J, Tian X (2020) SIRT3-mediated deacetylation of PRDX3 alleviates mitochondrial oxidative damage and apoptosis induced by intestinal ischemia/reperfusion injury. Redox Biol 28:101343

    Article  CAS  PubMed  Google Scholar 

  • Xu M, Li XY, Song L, Tao C, Fang J, Tao L (2021) miR-484 targeting of Yap1-induced LPS-inhibited proliferation, and promoted apoptosis and inflammation in cardiomyocyte. Biosci Biotechnol Biochem 85:378–385

    Article  PubMed  Google Scholar 

  • Yang X, Xu S, Qian Y, Xiao Q (2017) Resveratrol regulates microglia M1/M2 polarization via PGC-1alpha in conditions of neuroinflammatory injury. Brain Behav Immun 64:162–172

    Article  CAS  PubMed  Google Scholar 

  • Yoo S, Kim J-B (2015) Anti-apoptotic and beneficial metabolic activities of resveratrol in type II Gaucher disease. Biol Pharm Bull 38:913–918

    Article  CAS  PubMed  Google Scholar 

  • Yu D, Xiong J, Gao Y, Li J, Zhu D, Shen X, Sun L, Wang X (2021) Resveratrol activates PI3K/AKT to reduce myocardial cell apoptosis and mitochondrial oxidative damage caused by myocardial ischemia/reperfusion injury. Acta Histochem 123:151739

    Article  CAS  PubMed  Google Scholar 

  • Zaatout N (2022) An overview on mastitis-associated Escherichia coli: pathogenicity, host immunity and the use of alternative therapies. Microbiol Res 256:126960

    Article  CAS  PubMed  Google Scholar 

  • Zhang J, Dong X, Yan Q, Ren W, Zhang R, Jiang X, Geng Z, Xu X, Liu C, Zhang S, Liu D, Liu Y (2021) Galectin-1 inhibited LPS-induced autophagy and apoptosis of human periodontal ligament stem cells. Inflammation 44:1302–1314

    Article  CAS  PubMed  Google Scholar 

  • Zu Y, Chen XF, Li Q, Zhang ST, Si LN (2021) PGC-1alpha activates SIRT3 to modulate cell proliferation and glycolytic metabolism in breast cancer. Neoplasma 68:352–361

    Article  CAS  PubMed  Google Scholar 

Download references

Acknowledgements

We thank Prof. Wei Yang and Prof. Xudong Sun at Heilongjiang Bayi Agricultural University (Daqing, China) for providing the MAC-T as a gift.

Funding

This research was supported by a grant from the science and technology innovation program of Hunan Province (Changsha, China, 2022RC1161), the science fund for distinguished young scholars of Changsha City, China (Changsha, China, kq2106045), the Natural Science Foundation of Hunan (Changsha, China, 2022JJ40169), and the earmarked Fund for China Agriculture Research System (Beijing, China, CARS-37).

Author information

Authors and Affiliations

  1. College of Veterinary Medicine, Hunan Agricultural University, Changsha, 410128, China

    Lu Ouyang, Huilun Tang, Zilin Liu, Yuan Tian, Xin Gao, Tao Peng & Lei Liu

  2. College of Animal Science, Hunan Agricultural University, Changsha, 410128, China

    Zuo Wang, Xinyi Lan, Weijun Shen, Dingfu Xiao & Fachun Wan

Authors
  1. Lu Ouyang
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Huilun Tang
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Zilin Liu
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Yuan Tian
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Xin Gao
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Tao Peng
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Zuo Wang
    View author publications

    You can also search for this author in PubMed Google Scholar

  8. Xinyi Lan
    View author publications

    You can also search for this author in PubMed Google Scholar

  9. Weijun Shen
    View author publications

    You can also search for this author in PubMed Google Scholar

  10. Dingfu Xiao
    View author publications

    You can also search for this author in PubMed Google Scholar

  11. Fachun Wan
    View author publications

    You can also search for this author in PubMed Google Scholar

  12. Lei Liu
    View author publications

    You can also search for this author in PubMed Google Scholar

Contributions

Conceptualization, L.L. and W.F.; Methodology, O.L., H.T. and L.L.; Validation, O.L., H.T. and Z.L.; Formal Analysis, O.L., H.T., L.L.; Investigation, O.L., H.T., Z.L., Y.T., X. G., T.P. and Z.W.; Resources, L.L.; Data Curation, X.L.; Writing – Original Draft Preparation, L.L. and O.L.; Writing – Review & Editing, F.W., D. X. and L.L.; Visualization, X.L. and W.S.; Supervision, F.W. and L.L.; Project Administration, W.S.; Funding Acquisition, L.L., W.F. and D.X.

Corresponding authors

Correspondence to Dingfu Xiao, Fachun Wan or Lei Liu.

Ethics declarations

Conflicts of interest

The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.

Institutional review board statement

Not applicable.

Rights and permissions

Springer Nature or its licensor (e.g. a society or other partner) holds exclusive rights to this article under a publishing agreement with the author(s) or other rightsholder(s); author self-archiving of the accepted manuscript version of this article is solely governed by the terms of such publishing agreement and applicable law.

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Ouyang, L., Tang, H., Liu, Z. et al. Resveratrol inhibits LPS-induced apoptosis in bovine mammary epithelial cells: the role of PGC1α-SIRT3 axis. In Vitro Cell.Dev.Biol.-Animal 59, 264–276 (2023). https://doi.org/10.1007/s11626-023-00766-2

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s11626-023-00766-2

Keywords

Search

Navigation