Modulation of Cellular Senescence in HEK293 and HepG2 Cells by Ultrafiltrates UPla and ULu Is Partly Mediated by Modulation of Mitochondrial Homeostasis under Oxidative Stress

Zhou, Junxian; Liu, Kang; Bauer, Chris; Bendner, Gerald; Dietrich, Heike; Slivka, Jakub Peter; Wink, Michael; Wong, Michelle B. F.; Chan, Mike K. S.; Skutella, Thomas

doi:10.3390/ijms24076748

Open AccessArticle

Modulation of Cellular Senescence in HEK293 and HepG2 Cells by Ultrafiltrates UPla and ULu Is Partly Mediated by Modulation of Mitochondrial Homeostasis under Oxidative Stress

by

Junxian Zhou

¹,

Kang Liu

²,

Chris Bauer

³,

Gerald Bendner

¹

,

Heike Dietrich

¹,

Jakub Peter Slivka

⁴,

Michael Wink

⁵

,

Michelle B. F. Wong

⁶,

Mike K. S. Chan

⁶ and

Thomas Skutella

^1,*

¹

Institute for Anatomy and Cell Biology, Medical Faculty, Heidelberg University, 69120 Heidelberg, Germany

²

Department of Pharmacology of Chinese Materia Medica, China Pharmaceutical University, Nanjing 210009, China

³

MicroDiscovery, 10405 Berlin, Germany

⁴

Reviva s.r.o. Pekarska 6, 158 00 Prague, Czech Republic

⁵

Institute of Pharmacy and Molecular Biotechnology, Heidelberg University, 69120 Heidelberg, Germany

⁶

EW European Wellness International GmbH, 72184 Eutingen im Gäu, Germany

^*

Author to whom correspondence should be addressed.

Int. J. Mol. Sci. 2023, 24(7), 6748; https://doi.org/10.3390/ijms24076748

Submission received: 11 March 2023 / Revised: 31 March 2023 / Accepted: 3 April 2023 / Published: 4 April 2023

(This article belongs to the Special Issue Cellular Senescence in Physiological and Pathological Processes 2.0)

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Abstract

Protein probes, including ultrafiltrates from the placenta (UPla) and lung (ULu) of postnatal rabbits, were investigated in premature senescent HEK293 and HepG2 cells to explore whether they could modulate cellular senescence. Tris-Tricine–PAGE, gene ontology (GO), and LC–MS/MS analysis were applied to describe the characteristics of the ultrafiltrates. HEK293 and HepG2 cells (both under 25 passages) exposed to a sub-toxic concentration of hydrogen peroxide (H₂O₂, 300 μM) became senescent; UPla (10 μg/mL), ULu (10 μg/mL), as well as positive controls lipoic acid (10 μg/mL) and transferrin (10 μg/mL) were added along with H₂O₂ to the cells. Cell morphology; cellular proliferation; senescence-associated beta-galactosidase (SA-β-X-gal) activity; expression of senescence biomarkers including p16 INK4A (p16), p21 Waf1/Cip1 (p21), HMGB1, MMP-3, TNF-α, IL-6, lamin B1, and phospho-histone H2A.X (γ-H2AX); senescence-related gene expression; reactive oxygen species (ROS) levels; and mitochondrial fission were examined. Tris-Tricine–PAGE revealed prominent detectable bands between 10 and 100 kDa. LC–MS/MS identified 150–180 proteins and peptides in the protein probes, and GO analysis demonstrated a distinct enrichment of proteins associated with “extracellular space” and “proteasome core complex”. UPla and ULu modulated senescent cell morphology, improved cell proliferation, and decreased beta-galactosidase activity, intracellular and mitochondrial ROS production, and mitochondrial fission caused by H₂O₂. The results from this study demonstrated that UPla and Ulu, as well as lipoic acid and transferrin, could protect HEK293 and HepG2 cells from H₂O₂-induced oxidative damage via protecting mitochondrial homeostasis and thus have the potential to be explored in anti-aging therapies.

Keywords: cellular senescence; cell proliferation; SA-β-X-gal; senescence marker; intracellular ROS; mitochondrial ROS; mitochondrial fission

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MDPI and ACS Style

Zhou, J.; Liu, K.; Bauer, C.; Bendner, G.; Dietrich, H.; Slivka, J.P.; Wink, M.; Wong, M.B.F.; Chan, M.K.S.; Skutella, T. Modulation of Cellular Senescence in HEK293 and HepG2 Cells by Ultrafiltrates UPla and ULu Is Partly Mediated by Modulation of Mitochondrial Homeostasis under Oxidative Stress. Int. J. Mol. Sci. 2023, 24, 6748. https://doi.org/10.3390/ijms24076748

AMA Style

Zhou J, Liu K, Bauer C, Bendner G, Dietrich H, Slivka JP, Wink M, Wong MBF, Chan MKS, Skutella T. Modulation of Cellular Senescence in HEK293 and HepG2 Cells by Ultrafiltrates UPla and ULu Is Partly Mediated by Modulation of Mitochondrial Homeostasis under Oxidative Stress. International Journal of Molecular Sciences. 2023; 24(7):6748. https://doi.org/10.3390/ijms24076748

Chicago/Turabian Style

Zhou, Junxian, Kang Liu, Chris Bauer, Gerald Bendner, Heike Dietrich, Jakub Peter Slivka, Michael Wink, Michelle B. F. Wong, Mike K. S. Chan, and Thomas Skutella. 2023. "Modulation of Cellular Senescence in HEK293 and HepG2 Cells by Ultrafiltrates UPla and ULu Is Partly Mediated by Modulation of Mitochondrial Homeostasis under Oxidative Stress" International Journal of Molecular Sciences 24, no. 7: 6748. https://doi.org/10.3390/ijms24076748

APA Style

Zhou, J., Liu, K., Bauer, C., Bendner, G., Dietrich, H., Slivka, J. P., Wink, M., Wong, M. B. F., Chan, M. K. S., & Skutella, T. (2023). Modulation of Cellular Senescence in HEK293 and HepG2 Cells by Ultrafiltrates UPla and ULu Is Partly Mediated by Modulation of Mitochondrial Homeostasis under Oxidative Stress. International Journal of Molecular Sciences, 24(7), 6748. https://doi.org/10.3390/ijms24076748

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Modulation of Cellular Senescence in HEK293 and HepG2 Cells by Ultrafiltrates UPla and ULu Is Partly Mediated by Modulation of Mitochondrial Homeostasis under Oxidative Stress

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