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Using Mouse Mammary Tumor Cells to Teach Core Biology Concepts: A Simple Lab Module
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Effect of matrine in MAC-T cells and their transcriptome analysis: A basic study.

Zhao Zhang1, Yuze Yang2, Lijiao Yan1

  • 1College of Veterinary Medicine, Gansu Agricultural University, Lanzhou, China.

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Matrine is not toxic to bovine mammary epithelial cells (MAC-T) at 0.5 mg/mL but induces apoptosis at 2 mg/mL. This study clarifies matrine

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Area of Science:

  • Pharmacology
  • Cell Biology
  • Toxicology

Background:

  • Matrine, a herbal alkaloid, exhibits antibacterial properties and is a potential antibiotic alternative for mastitis treatment.
  • Antibiotic restrictions necessitate exploring safe and effective alternatives like matrine.
  • The safe concentration and toxicological mechanisms of matrine on bovine mammary epithelial cells (MAC-T) remain unclear.

Purpose of the Study:

  • To determine the safe concentration of matrine on MAC-T cells.
  • To investigate matrine-induced apoptosis and its time-dependent effects.
  • To elucidate the molecular mechanisms underlying matrine's toxicity and apoptotic effects in MAC-T cells.

Main Methods:

  • MAC-T cells were treated with varying matrine concentrations (0.5–3 mg/mL) for different durations (4–24 h).
  • Lactate dehydrogenase (LDH) assay was used to assess cytotoxicity.
  • Flow cytometry detected apoptosis, and RNA-sequencing (RNA-seq) analyzed gene expression changes.
  • Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis identified affected biological pathways.

Main Results:

  • Matrine showed no cytotoxicity at 0.5 mg/mL but was cytotoxic at concentrations from 1 to 3 mg/mL.
  • Matrine induced apoptosis in MAC-T cells at 2 mg/mL, with increasing apoptosis over time.
  • RNA-seq identified 1645 differentially expressed genes (DEGs), with 676 upregulated and 969 downregulated.
  • KEGG analysis linked matrine toxicity and apoptosis to pathways including cytokine-cytokine receptor interaction and the P53 pathway.
  • Seven DEGs were identified as potentially associated with matrine-induced toxicity and apoptosis.

Conclusions:

  • Matrine is safe at concentrations up to 0.5 mg/mL for MAC-T cells.
  • Matrine at 2 mg/mL induces time-dependent apoptosis in MAC-T cells.
  • Understanding matrine's molecular mechanisms provides a basis for its safe application in mastitis prevention and treatment.