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Knockdown MiR-302b Alleviates LPS-Induced Injury by Targeting Smad3 in C28/I2 Chondrocytic Cells

Yueshu Wang1, Tao Yu2, Hui Jin3

  • 1Department of Hand Surgery, China-Japan Union Hospital of Jilin University, Changchun, China.

Cellular Physiology and Biochemistry : International Journal of Experimental Cellular Physiology, Biochemistry, and Pharmacology
|February 8, 2018
PubMed
Abstract

Insights

MicroRNA-302b (miR-302b) exacerbates inflammation and apoptosis in osteoarthritis chondrocytes. Suppressing miR-302b protects against these effects by up-regulating Smad3, offering a potential therapeutic strategy for osteoarthritis.

Area of Science:

  • Molecular Biology
  • Cell Biology
  • Biochemistry

Background:

  • Osteoarthritis (OA) is a prevalent degenerative joint disease.
  • MicroRNAs (miRNAs) play a role in OA pathogenesis, but miR-302b's function is unclear.
  • This study investigates miR-302b's role in lipopolysaccharide (LPS)-induced chondrocyte injury.

Purpose of the Study:

  • To elucidate the role of miR-302b in LPS-induced chondrocyte injury.
  • To identify miR-302b target proteins involved in chondrocyte inflammation and apoptosis.
  • To explore miR-302b's potential as a therapeutic target in OA.

Main Methods:

  • Human OA chondrocytes (C28/12 cell line) were used.
  • Cells were transfected with miR-302b inhibitors and mimics.
  • LPS treatment was applied to induce injury.
  • Cell viability, apoptosis, and inflammatory cytokine expression were assessed.
  • Dual luciferase assays identified miR-302b targets.
  • Western blotting analyzed protein expression of Smad3, Notch, and mTOR signaling pathways.

Main Results:

  • LPS treatment reduced chondrocyte viability, increased apoptosis, and elevated inflammatory cytokines.
  • miR-302b expression was upregulated in LPS-treated chondrocytes.
  • miR-302b mimic transfection worsened LPS effects; miR-302b inhibition reversed them.
  • Smad3 was identified as a direct target of miR-302b, with miR-302b negatively regulating its expression.
  • miR-302b inhibition suppressed inflammation by up-regulating Smad3.
  • miR-302b inhibition increased Notch and mTOR signaling pathway proteins, with miR-302b directly targeting Notch2.

Conclusions:

  • miR-302b aggravates LPS-induced chondrocyte injury, promoting apoptosis and inflammation.
  • miR-302b directly targets and down-regulates Smad3 and Notch2.
  • Suppression of miR-302b may protect against OA progression by up-regulating Smad3 and modulating Notch/mTOR pathways.
Keywords:
ChondrocytesMiR-302bOsteoarthritisSmad3

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